Since the diccovery of neural stem cells(NSCs)in the embryonic and adult mammalian central nerous system,it provided novel ideas forneurogenesis as the potential of proliferation and differentiation of NSCs.One of the...Since the diccovery of neural stem cells(NSCs)in the embryonic and adult mammalian central nerous system,it provided novel ideas forneurogenesis as the potential of proliferation and differentiation of NSCs.One of the ways to promote the clinical application of neural stem cells(NSCs)is searching effective methods which regulate the proliferation and differentiation.This is also a problem urgently to be solved in medical field.Plenty of earlier studies have shown that traditional chinese medicine can promote the proliferation and differentiation of NSCs by regulating the related signaling pathway in vivo and in vitro.The reports of Chinese and foreign literatures on regulating the proliferation and differentiation of neural stem cells in recent ten years and their target and signaling pathways is analyzed in this review.The traditional chinese medicine regulate proliferation and differentiation of NSCs by the signaling pathways of Notch,PI3K/Akt,Wnt/β-catenin,and GFs.And,those signaling pathways have cross-talk in the regulation progress.Moreover,some traditional Chinese medicine,such as astragalus,has a variety of active ingredients to regulate proliferation and differentiation of NSCs through different signaling pathways.However,to accelerate the clinical application of neural stem cells,the studies aboutthe proliferation and differentiation of NSCs and Chinese medicine should be further deepened,the mechanism of multiple targets and the comprehensive regulation function of traditional Chinese medicine should be clarified.展开更多
Objective Diabetic patients pose a greater challenge in managing chronic wound healing,leading to a higher amputation risk compared to non-diabetic patients.Due to their paracrine function by secreting various cytokin...Objective Diabetic patients pose a greater challenge in managing chronic wound healing,leading to a higher amputation risk compared to non-diabetic patients.Due to their paracrine function by secreting various cytokines and angiogenic factors,mesenchymal stem cells(MSCs)have been acknowledged to be a potential agent in modulating wound healing process.However,post-transplanted MSCs are vulnerable to death,indicating poor survival and migration ability in the wound site of the host,especially under hyperglycemia.As hyperglycemia induces reactive oxygen species(ROS)generation and cellular apoptosis,improvement of MSCs survival and migration potentials under hyperglycemia could contribute to a more efficient MSCs-based wound healing therapy.Salidroside(Sa),a small-molecule drug derived from Rhodiola plant,has been proved to enhance the paracrine function of skeletal muscle cells,as well as their migration even under hypoxichyperglycemia.Herein,we investigated whether Sa could improve the survival and migration potentials of MSCs,subsequently enhance the wound healing process under hyperglycemia.Methods MSCs were cultured under three conditions:low glucose,high glucose,and high glucose+Sa.qPCR analysis and western blotting were done to examine the mRNA and protein expression level of several factors which are important in upregulating the wound healing process.MTT colorimetric assay,intracellular ROS detection,and flow cytometry assay were employed to examine the effect of Sa in MSCs survival.Transwell chamber assay,scratch assay,and phalloidin staining were done to elucidate the role of Sa in regulating MSCs migration potential.For in vivo experiment,diabetic wound healing mice model was generated to elucidate the effect of Sa-pretreated MSCs transplantation in wound closure rate,as well as re-epithelization status,observed with hematoxylin and eosin staining.The diabetic wound healing mice model were divided into three groups:1)mice injected with PBS,2)mice transplanted with PBS-pretreated MSCs,and 3)mice transplanted with Sa-pretreated MSCs.Results(1)Hyperglycemic condition induced the generation of ROS and suppressed total cell number of MSCs,while Sa treatment into MSCs restored these hyperglycemia-induced alterations.In line with this,total apoptotic cells were also suppressed by treating MSCs with Sa.The expression level of cell survival factor,heme-oxygenase 1(HO-1),was enhanced in Sa-pretreated MSCs.Further treatment of HO-1 inhibitor into Sa-pretreated MSCs nullified the ROS level and total apoptotic cells,indica-ting the importance of HO-1 in mediating the Sa-induced survival of MSCs under hyperglycemia.(2)Transwell chamber and scratch assay results showed that Sa-pretreated MSCs have a higher migration potential under hyperglycemia,supported by higher F-actin polymerization fractal dimension.Fibroblast growth factor 2(FGF2)and hepatocyte growth factor(HGF)expression level,which are essential factors for cell migration,were also improved in Sa-pretreated MSCs under hyperglycemia.(3)In diabetic wound healing mice model,transplantation of Sa-pretreated MSCs resulted in significantly improved wound closure rate and re-epithelization.The protein levels of HO-1,FGF2,and HGF were also enhanced in the tissues obtained from the wound site of diabetic wound healing mice model which were transplanted with Sa-pretreated MSCs.Conclusions Salidroside pretreatment on MSCs could improve their survival and migration potentials,subsequently promoting wound healing process under hyperglycemia.This prospective MSC-based therapy could serve as a novel strategy to improve diabetic wound healing.展开更多
Cancer stem cells(CSCs)are the driving force for sustainable tumor growth and metastasis and responsible for drug resistance and cancer relapse.Nanoparticle-based drug delivery has been demonstrated to be effective in...Cancer stem cells(CSCs)are the driving force for sustainable tumor growth and metastasis and responsible for drug resistance and cancer relapse.Nanoparticle-based drug delivery has been demonstrated to be effective in combating tumor growth.However,it has been challenging to selectively eliminate CSCs due to the lack of a general signature for a spectrum of cancers.It is known that CSCs from various types of cancer show lower stiffness compared to non-CSCs.It remains unclear whether low stiffness in CSCs influences cellular uptake in nanoparticle-based drug delivery and thus the chemotherapy efficacy.Graphene quantum dot(GQD)is emerging as a promising carrier material in delivering anti-cancer drugs.We found that breast CSCs were softer than conventional cancer cells,which were further softer compared to healthy breast tissue cells.Importantly,soft CSCs uptook more GQD than conventional cancer cells,while stiff breast cancer cells with relatively low stiffness uptook more GQD than healthy breast cells.Softening cells by pharmacologically inhibiting actomyosin activity using either siRNA or actomyosin inhibitors significantly enhanced the cellular uptake of GQD in breast cancer cells but not CSCs,while stiffening cells by activating actomyosin using CA-MLCK/ROCK or actomyosin activators considerably suppressed the nanoparticle uptake in both cancer cells and CSCs.GQD could specifically target CSC because of low cell stiffness of CSC in breast cancer cell line MCF-7 and MDA-MB-231.Further regulating cell stiffness reflected that decreasing breast cancer cell stiffness by inhibiting actomyosin activity using blebbistatin could promote GQD uptake.Vice versa,stiffening cancer cell by activating actomyosin decreased GQD uptake.The attachment of anti-cancer drug doxorubicin did not alter the trend of GQD uptake in neither soften nor stiffen cancer cells.Actomyosin activity regulates cellular uptake ofGQD might through clathrin and caveolin-mediated endocytosis.Cancer cells are softer than normal cells from the same organ,CSC are softer than non-CSC.Thus we further confirmed that the GQD uptake of normal breast cell line MCF-10 is less than breast cancer cell line MCF-7 and MDA-MB-231.Suggesting the clinical significance that using GQD as drug carrier targeting softer cells could reduce side effects to normal tissue cells.Since CSC are softer than non-CSC,GQD could decreased the percentage of CSC in whole cancer cell population by targeting softer cells.These results suggesting that it would be possible to target cancer cells and CSC by targeting from a perspective of cell mechanical difference.High uptake of nanoparticles in soft cancer cells could not be explained by their differential membrane potentials.Mechanistically,low cell mechanics or inhibiting actomyosin activity activated both clathrin and caveolin-mediated endocytosis signaling pathways,while high cell mechanics or activating actomyosin suppressed these signalings.Pharmacologically inhibiting clathrin or caveolin-mediated endocytosis signaling significantly decreased GQD uptake in CSCs and in conventional breast cancer cells when actomyosin was suppressed.Further,GQD conjugated with doxorubicin could be specifically delivered into CSCs with low stiffness and eliminated more CSCs in the presence of both CSCs and non-CSCs.Taken together,these data reveal the regulatory role of cell mechanics in cellular uptake of nanoparticles and demonstrate that GQD can be utilized to specifically eliminate CSCs,which have important implications in nanoparticle-based drug delivery for cancer therapy.展开更多
OBJECTIVE To establish an in vitro cel model based on patient-specific human microglia to study the pathological mechanism of Alzheimer disease(AD) and to screen candidate drugs.METHODS First,the induced pluripotent s...OBJECTIVE To establish an in vitro cel model based on patient-specific human microglia to study the pathological mechanism of Alzheimer disease(AD) and to screen candidate drugs.METHODS First,the induced pluripotent stem cells(iPSCs) of AD patients and cognitive normal controls(CNC) were induced to hematopoietic progenitor cells(HPCs),and then HPCs were further induced with IL-34,M-CSF,GM-CSF and TGF-β1 for 20 d to obtain microglialike cells(MGLCs).HPCs were isolated by flow cytometry and MGLCs were identified by immunofluorescence.Cell phagocytosis was determined by phagocytosis neutral red experiusing Luminex assay kits,and the cell growth curve during the experiment was recorded by IncuCyte ZOOM.The phagocytic ability and secretion of cytokines of MGLCs were observed under the stimulation of LPS.RESULTS MGLCs from AD patients(AD-MGLCs) and CNC expressed microglia markers IBA1,TMEM119,P2 RY12,TREM2 and CD11 B.The results of phagocytosis neutral red experiment showed that under normal conditions,AD-MGLCs had stronger phagocytic ability(P<0.01).Stimulation by LPS resulted in increased phagocytosis of cel s,and the increase in phagocytosis of CNC-MGLCs was higher than AD-MGLCs(P<0.01).Experiments showed that high concentrations of LPS(>2 mg·L^(-1)) resulted in CNC-microglia death(P<0.01),whereas ADMGLCs did not show significant death.The cytokine assay showed that under normal conditions,the concentrations of IFN-γ and IL-2 secreted by AD patients were slightly higher than those of CNC.After LPS stimulation,the secretion of TNF-α,IL-6 and IL-10 was significantly increased.The increased secretion of AD-MGLCs was greater than that CNC-MGLCs(P<0.01).CONCLUSION AD-iPSCs derived MGLCs exhibit significant inflammatory characteristics and are more active than CNC,which may be associated with chronic inflammatory responses caused by microglia in AD,thus may provide valuable new tools to screen candidate drugs for the disease and to discover the mechanisms underlying AD pathogenesis.展开更多
OBJECTIVE To investigate the inhibitory effect of scutellarin on the self-renewal and differentiation of HT-29 cells-derived cancer stem-like cells(HT-29CSC)in vitro and in vivo,and to explore its mechanism.METHODS Th...OBJECTIVE To investigate the inhibitory effect of scutellarin on the self-renewal and differentiation of HT-29 cells-derived cancer stem-like cells(HT-29CSC)in vitro and in vivo,and to explore its mechanism.METHODS The effect of scutellarin on the growth of HT-29CSC was determined by 3D Culture assay.The effect of scutellarin on growth and transformation of HT-29CSC was probed by soft agar colony formation assay.The effect of scutellarin on the differentiation of HT-29CSC was determined by serum induction differentiation assay in vitro.The effects of scutellarin on the expressions of marker gene Lgr5,target gene c-Myc,proliferation gene CK20 and Nanog gene were measured by quantitative real-time RT-PCR.Investigate the effect of scutellarin on the expression of c-Myc,Gli1,and Lgr5 protein by Western blotting.A subcutaneous xenograft model of colon cancer in nude mice was established and administered by intraperitoneal injection.The change of body weight and tumor size of nude mice were observed every two days.Investi⁃gate the effects of scutellarin on the growth of xenograft tumors in nude mice.The expression of CD133,Lgr5,Gli1,Ptch1,c-Myc,Ki67,CK20,Nanog gene in tumors were measured by quantitative real-time RT-PCR.The expression of c-Myc,Gli1,Lgr5,CD133,Ki67 protein were measured by Western blotting.RESULTS Scutellarin can inhibit the growth of HT-29CSC in 3D culture.Compared with the solvent control group,scutellarin can significantly inhibit the growth and transformation and differentiation of HT-29CSC in vitro(P<0.01).The expression levels of marker genes Lgr5,target gene c-Myc,proliferation gene CK20 and Nanog in HT-29CSC were down-regulated by scutellarin.Scutellarin can reduce the expression of c-Myc,Gli1,and Lgr5 protein in HT-29CSC.Scutellarin can inhibit the growth of colon cancer xenografts,lower CD133,Lgr5,Gli1,Ptch1,c-Myc,Ki67,CK20,and Nanog mRNA level of xenograft tumors,reduce the expression of c-Myc,Gli1,Lgr5,CD133,and Ki67 protein of xenograft tumors in nude mice.CONCLUSION Scutellarin,which is the main component of scutellaria barbata,can inhibit the differentiation of HT-29CSC and the mechanism is to inhibit the activity of Hedgehog signaling pathway.展开更多
The experiment was designed to study effects of retinoic acid and ascorbic acid on differentiation of mouse embryonic stem cells to cardiomyocytes. Embryonic bodies （EB） were developed from mESC in suspension cultur...The experiment was designed to study effects of retinoic acid and ascorbic acid on differentiation of mouse embryonic stem cells to cardiomyocytes. Embryonic bodies （EB） were developed from mESC in suspension culture, different levels of concentration of retinoic acid and ascorbic acid were used to determine the optimal conditions for EB formation. The results showed that the optimal concentrations were 10.9 mol. L-1 and 0.1 mg. mL-1 for retinoic acid and ascorbic acids, respectively. 50% of EB which was significantly （p〈0.05） different from the control group developed to cardiomyocytes. In conclusion, rctinoic acid and ascorbic acid had strong ability to promote cardiomyocyte differentiation of mouse embryonic stem cells. 10-9 mol. L-1 retinoic acid and 0.10 mg. mL-1 ascorbic acids were recommended to induce differentiation of mouse ES ceUs toward cardiomyocytes.展开更多
Non-alcoholic fatty liver disease(NAFLD)is the most common chronic liver disease,defined by several phases,ranging from benign fat accumulation to non-alcoholic steatohepatitis(NASH),which can lead to liver cancer and...Non-alcoholic fatty liver disease(NAFLD)is the most common chronic liver disease,defined by several phases,ranging from benign fat accumulation to non-alcoholic steatohepatitis(NASH),which can lead to liver cancer and cirrhosis.Although NAFLD is a disease of disordered metabolism,it also involves several immune cell-mediated inflammatory processes,either promoting and/or suppressing hepatocyte inflammation through the secretion of pro-inflammatory and/or anti-inflammatory factors to influence the NAFLD process.However,the underlying disease mechanism and the role of immune cells in NAFLD are still under investigation,leaving many open-ended questions.In this review,we presented the recent concepts about the interplay of immune cells in the onset and pathogenesis of NAFLD.We also highlighted the specific non-immune cells exhibiting immunological properties of therapeutic significance in NAFLD.We hope that this review will help guide the development of future NAFLD therapeutics.展开更多
Objective The binding of cell adhesive peptides(such as RGD)to integrins initiates the recruitment of cytoplasmic adaptor proteins(e.g.,vinculin)and the formation of focal adhesion(FA)complexes required for cell adhes...Objective The binding of cell adhesive peptides(such as RGD)to integrins initiates the recruitment of cytoplasmic adaptor proteins(e.g.,vinculin)and the formation of focal adhesion(FA)complexes required for cell adhesion.The ability to manipulate this ligand-mediated cell adhesion process is crucial for regulating cell migration,cell differentiation,injury healing,and immune response.Some recent studies reported the importance of the tether length/mobility of the cell adhesive ligands in regulating the traction force development of cells.In the native cellular microenvironment,such a dynamic change in the nanoscale tether length of bioactive ligands is often mediated by conformational changes of the structural proteins due to protein folding or degradation.However,no prior studies have demonstrated the modulation of the ligand tether mobility by controlling the intramolecular folding of polymeric linkers.Unfoldable synthetic macromolecules with easy synthetic routes and controllable structures,such as supramolecular host-guest single chain nanogels(SCNGs),are ideal candidates for mimicking the changes in the tether mobility of bioactive ligands via biorthogonal triggers.Methods S,S’-bis(a’a’-dimethyl-a’’-propargyl acetate)trithiocarbonate was first used to mediate the RAFT polymerization of N,N-dimethyl acrylamide,vinyl-adamantane and vinyl-β-cyclodextrin to yield the ADA@CD-SCNGs.The preparation of the unfoldable host-guest SCNGs was evidenced by the by gel permeation chromatography,proton nuclear magnetic resonance spectroscopy,atomic force microscopy and dynamic light scattering.Then the RGD peptide was conjugated to the alkynyl group on one end of the SCNGs before immobilizing the material on the substrate,which was confirmed by scanning electron microscopy(SEM).The regulation of cell behaviours by unfolding of the SCNG-RGD was confirmed by immunofluorescence staining of vinculin and Yes-associated protein(YAP).Results The preparation of ADA@CD-SCNGs was confirmed by GPC which showed a unimodal molecular weight distribution.DLS and AFM data also proved that the SCNGs had an average diameter of 12±3nm.SEM images showed that SCNGs were conjugated as a linker of RGD peptide to thiolated glass substrate at an average density of 162±11 particles/μm2.These particles disappeared after adding free competitive ADA guest molecules,indicating the triggered unfolding of the tether SCNGs.In addition,the unfolding of supramolecular ADA@CD-SCNGs was also evidenced by a decrease in the GPC elution time and a slight increase in the apparent molecular weight.These results show that the immobilized ADA@CD-SCNGs can be unfolded to tune the tether length and mobility of the conjugated RGD ligands.Then we investigated the regulation of the cell behaviors on the substrate by triggering the unfolding of SCNG linkers.A critical level of traction force is required to effectively initiate and maintain integrin-mediated formation of FA complexes and subsequent mechano-transduction signaling.An increased tether length in cell-adhesive ligands can lead to a diminished cell traction force as if cells are adhering to soft substrates.Here,the unfolding of the ADA@CD-SCNG-RGD triggered by the addition of free ADA led to disassembly of the mature focal adhesions in the cells as evidenced by the reduced vinculin and F-actin in staining.Subsequently,nuclear YAP also decreased significantly because of the impaired mechano-sensing and diminished cell cytoskeleton tension.In addition,the extensively spread cells gradually became round after the medium was supplemented with free competitive ADA to unfold the SCNG linker.These finding demonstrates that the substrates with the unfolded ADA@CD-SCNG-RGD only supported weak cell adhesions.In contrast,on the substrate conjugated with the nonunfoldable MBA-SCNG-RGD linker,the addition of free ADA resulted in no change in the spread cell morphology and protein expressions.These results indicate that the unfoldable host-guest ADA@CD-SCNG can be used to manipulate the nanoscale presentation of ligands to regulate cell behaviors.Conclusions We demonstrate the application of SCNGs as the supramolecular linker to tune the nanoscale ligand tether length.These findings demonstrate that the strategy of manipulating the tether mobility of bioactive ligands by using supramolecular SCNGs as linkers provides a highly tunable,biomimetic,and bio-orthogonal approach to study the dynamic events of cell adhesion.展开更多
Aim The present study aims to investigate whether BMSCs transplantation may inhibit hypertrophic hearts and its underlying mechanisms. Background There is no evidence so far that Bone marrow-derived mesenchy- mal stem...Aim The present study aims to investigate whether BMSCs transplantation may inhibit hypertrophic hearts and its underlying mechanisms. Background There is no evidence so far that Bone marrow-derived mesenchy- mal stem cells （BMSCs） can heal pathological myocardial hypertrophy. Methods To observe the antihypertrophic actions, BMSCs was indirectly cocultured with NRVCs in vitro, or intramyocardially transplanted into hypertrophic hearts in vivo. Results ISO-induced typical hypertrophic characteristics of cardiomyocytes were obviously preven- ted by BMSCs in the co-culture model in vitro and after BMSCs transplantation in vivo. Furthermore, the activation of the Ca2＋/calcineurin/NFATc3 hypertrophic pathway was shown abrogated in the presence of BMSCs both in vitro and in vivo. Interestingly, blockage of VEGF release from BMSCs but not bFGF and IGF-1 can abolish the protec- tive effects of BMSCs on cardiomyocytes hypertrophy. Consistently, VEGF administration attenuated ISO-induced BNP and β-MHC expression and the activation of Ca2＋/cal- the enlargement of cellular size, the augment of ANP, cineurin/NFATc3 hypertrophic pathway, and these can be abrogated by blocking VEGFR-1, indicating VEGFR-1 is involved in the antihypertrophic role of VEGF. We further find that the ample VEGF secretion contributing to the anti-hypertrophic effects of BMSCs originates from BMSCs interplay with cardiac cells but not BMSCs or cardiomyo- cytes alone. Conclusions Thus, mesenchymal stem cells are able to inhibit myocardial hypertrophy via interacting with cardiomyocytes so as to promote VEGF release which inhibits the activation of the Ca2＋/calcineurin/NFATc3 hypertrophic signaling pathway in cardiac cells, in addition to its well-recognized ability to ameliorate myocardial injuries by replacing dead cells.展开更多
Embryonic stem (ES) cell biology is attracting much attention in cell biology because of their pluripotent behaviors and potential therapeutic applications. However,what maintains ES cell pluripotency and what trigger...Embryonic stem (ES) cell biology is attracting much attention in cell biology because of their pluripotent behaviors and potential therapeutic applications. However,what maintains ES cell pluripotency and what triggers ES cell展开更多
Spinal cord injury repair is one of the major challenges in medicine,as it can lead to permanent loss of function of central nervous system and damage to other function of the body.Stem cell transplantation together w...Spinal cord injury repair is one of the major challenges in medicine,as it can lead to permanent loss of function of central nervous system and damage to other function of the body.Stem cell transplantation together with tissue engineering is increasingly becoming a potential choice of treatment.However,direct transplantation of stem cells without scaffolds has yielded poor clinical outcome.Here we show a strategy of using mouse embryonic stem cells(ESCs)cultured within a silk fibroin(SF)based,three-dimensional scaffold with oriented channels by a directional temperature field freezing technique and lysophilization.We find that the ESCs maintained proliferation and migrated in the scaffolds and the cells migrated fastest along the SF channels.SF scaffolds contributed to ESC differentiation into neural and glial cell like cells and expressions of the neural and glial cell markers MAP2 and GFAP were greatly elevated when retinoic acid was used as an inducing factor.Our results suggest that this approach may offer some hope in the future for spinal cord injury repair using SF scaffolds and ESCs.展开更多
OBJECTIVE To explore the key mechanism of Bazi Bushen capsule(BZBS)in delaying the senescence of mesenchymal stem cells(MSCs)through network pharmacology and in vitro experiments.METHODS Network pharmacology was used ...OBJECTIVE To explore the key mechanism of Bazi Bushen capsule(BZBS)in delaying the senescence of mesenchymal stem cells(MSCs)through network pharmacology and in vitro experiments.METHODS Network pharmacology was used to predict the mechanism targets of BZBS in delaying MSCs senescence.A MSCs senescence model induced by D-galactose(D-gal)was used to investigate the effect and mechanism of BZBS on MSCs senescence in vitro.RESULTS Network pharmacology analy⁃sis showed that BZSB could delay MSCs senes⁃cence.The experiment showed that BZBS could significantly improve the survival activity of the aged MSCs.It significantly reduced the positive rate ofβ-galactosidase staining and p16,p21 expression in aged MSCs,enhanced the ability of adipogenic differentiation and osteogenic differentiation,and increased expression of Nanog,OCT4 and SOX2 in senescent MSCs.CONCLU⁃SIONS Network pharmacology and in vitro cell experiments verified that BZBS could delay MSCs senescence.展开更多
Objective Bone-marrow stem-cell transplantation has been shown to improve cardiac function in patients with AMI, but the safety of intracoronory infusion of autologous peripheral blood stem-cell(PBSCs) in patients wit...Objective Bone-marrow stem-cell transplantation has been shown to improve cardiac function in patients with AMI, but the safety of intracoronory infusion of autologous peripheral blood stem-cell(PBSCs) in patients with AMI is unknown. For this reason, we observe the feasibility and safety of PBSCs transplantation by intracoronory infusion in such patients.Method Fourty one patients with AMI were allocated to receive Granulocyte Colony-Stimulating Factor (G-CSF:Filgrastim,300 μg) with the dose of 300 μg-600 μg/day to mobilize the stem cell, and the duration of applying G-CSF was 5 days . On the sixth day, PBSCs were separated by Baxter CS 3000 blood cell separator into suspend liquid 57 ml. Then the suspend liquid was infused into the infarct related artery (IRA)by occluding the over the wire balloon and infusing artery through balloon center lumen. In the process of the intracoronary infusion of PBSCs, the complications should be observed, which were arrhythmias including of bradycardia, sinus arrest or atrial ventricular block, premature ventricular beats ,ventricular tachycardia, ventricular fibrillation; and hypotention, etc. Results There were total 10 cases with complications during the intracoronary infusion of PBSCs. The incidence of complications was 24.4%(10/41), including bradycardia is 2.4 %(1/41), sinus arrest or atrial ventricular block is 4.9%(2/41), ventricular fibrillation is 2.4 %( 1/41), hypotention is14.6 % (6 /41).Conclusions In patients with AMI, intracoronary infusion of PBSCs is feasible and safe.展开更多
Aim Aging is an independent risk factor for heart disease, however the effective intervention has not been found so far. Bone marrow mesenchymal stem cells （BMSCs） have been shown to offer a wide variety of cel- l...Aim Aging is an independent risk factor for heart disease, however the effective intervention has not been found so far. Bone marrow mesenchymal stem cells （BMSCs） have been shown to offer a wide variety of cel- lular functions including the protective effects on damaged hearts. Here we investigated the antiaging properties of BMSCs and the underlying mechanism in a cellular model of cardiomyocyte senescence and a rat model of aging hearts. Methods In vitro study, neonatal rat ventricular cells （NRVCs） and BMSCs were cocultured in the same dish with a semipermeable membrane to separate the two populations. In vivo, the BMSCs were injected into the rat hearts to observe their antiaging effects. The expression of β-galactosidase and aging-related proteins, and the lev- els of oxidative stress were determined in vivo and in vitro. The heart function was measured by the High-Resolution Imaging System. Results Monocultured NRVCs displayed the senescence-associated phenotypes, characterized by an increase in the number of β-galaetosidase-positive cells and decreases in the degradation and disappearance of cellular organelles in a time-dependent manner. The levels of reactive oxygen species and malondialdehyde were el- evated, whereas the activities of antioxidant enzymes superoxide dismutase and glutathione peroxidase were de- creased, along with upregulation of p53, p21cipl/wafl and p16INK4a in the aging eardiomyoeytes. These deleterious alterations were abrogated in aging NRVCs cocultured with BMSCs. Qualitatively, the same senescent phenotypes were consistently observed in aging rat hearts. Notably, BMSC transplantation significantly prevented these detri- mental alterations and improved the impaired cardiac function in the aging rats. Conclusions BMSCs possess strong antiseneseence action on the aging NRVCs and hearts and can improve cardiac function after transplantation in aging rats. The present study, therefore, provides an alternative approach for the treatment of heart failure in the elderly population.展开更多
OBJECTIVE To establish an in vitro cell model based on patient-specific human neural stem cells to study the pathomechanism of sporadic AD as well as screen candidate drugs.METHODS The peripheral blood cells from spor...OBJECTIVE To establish an in vitro cell model based on patient-specific human neural stem cells to study the pathomechanism of sporadic AD as well as screen candidate drugs.METHODS The peripheral blood cells from sporadic AD patients and cognitive normal controls were repro.grammed into inducedpluripotent stem cells(iPSCs),which were further induced into neural stem cells and neurons.The cell growth curve during the differentiation process was recorded by the IncuCyte ZOOM,and neural stem cells and neurons were identified by immunofluorescence.The apoptosis of neural stem cells and neurons was detected by Click-iT~Plus TUNEL Assay.RESULTS Neural stem cells derived from AD patients and cognitive normal controls can express neural stem cell markers Nes.tin,Sox1,Sox2 and Ki67.TUNEL assay results showed that the number of TUNEL-positive cells in neu.ral stem cells derived from AD patients was significantly higher than that of cognitive normal controls(P<0.01).When neural stem cells were differentiated into neurons,the percentage of MAP2 positive cells in the neural stem cell-derived culture dish of AD patients was significantly higher than the cogni.tive normal controls at day 16 of neuronal differentiation(P<0.01);the TUNEL assay showed that the number of TUNEL-positive cells in AD-derived neurons was significantly greater than that in cognitive normal controls(P<0.01) at day 16 of neuronal differentiation.CONCLUSION Our study revealed that AD-iPSC-derived neural stem cells exhibit premature neuronal differentiation and increased neural apoptosis,which might be relevant to the neuronal loss of AD,thus may provide valuable new tools to screen candidate drugs for the disease and to discover the mechanisms underlying AD pathogenesis.展开更多
基金supported by National Natural Science Foundation of China(81473549)Fundamental Research Funds for Central Universities(XDJK2017E158)
文摘Since the diccovery of neural stem cells(NSCs)in the embryonic and adult mammalian central nerous system,it provided novel ideas forneurogenesis as the potential of proliferation and differentiation of NSCs.One of the ways to promote the clinical application of neural stem cells(NSCs)is searching effective methods which regulate the proliferation and differentiation.This is also a problem urgently to be solved in medical field.Plenty of earlier studies have shown that traditional chinese medicine can promote the proliferation and differentiation of NSCs by regulating the related signaling pathway in vivo and in vitro.The reports of Chinese and foreign literatures on regulating the proliferation and differentiation of neural stem cells in recent ten years and their target and signaling pathways is analyzed in this review.The traditional chinese medicine regulate proliferation and differentiation of NSCs by the signaling pathways of Notch,PI3K/Akt,Wnt/β-catenin,and GFs.And,those signaling pathways have cross-talk in the regulation progress.Moreover,some traditional Chinese medicine,such as astragalus,has a variety of active ingredients to regulate proliferation and differentiation of NSCs through different signaling pathways.However,to accelerate the clinical application of neural stem cells,the studies aboutthe proliferation and differentiation of NSCs and Chinese medicine should be further deepened,the mechanism of multiple targets and the comprehensive regulation function of traditional Chinese medicine should be clarified.
基金Supported by grants from the National Natural Science Foundation of China ( 81372202,81872273, 31871367)
文摘Objective Diabetic patients pose a greater challenge in managing chronic wound healing,leading to a higher amputation risk compared to non-diabetic patients.Due to their paracrine function by secreting various cytokines and angiogenic factors,mesenchymal stem cells(MSCs)have been acknowledged to be a potential agent in modulating wound healing process.However,post-transplanted MSCs are vulnerable to death,indicating poor survival and migration ability in the wound site of the host,especially under hyperglycemia.As hyperglycemia induces reactive oxygen species(ROS)generation and cellular apoptosis,improvement of MSCs survival and migration potentials under hyperglycemia could contribute to a more efficient MSCs-based wound healing therapy.Salidroside(Sa),a small-molecule drug derived from Rhodiola plant,has been proved to enhance the paracrine function of skeletal muscle cells,as well as their migration even under hypoxichyperglycemia.Herein,we investigated whether Sa could improve the survival and migration potentials of MSCs,subsequently enhance the wound healing process under hyperglycemia.Methods MSCs were cultured under three conditions:low glucose,high glucose,and high glucose+Sa.qPCR analysis and western blotting were done to examine the mRNA and protein expression level of several factors which are important in upregulating the wound healing process.MTT colorimetric assay,intracellular ROS detection,and flow cytometry assay were employed to examine the effect of Sa in MSCs survival.Transwell chamber assay,scratch assay,and phalloidin staining were done to elucidate the role of Sa in regulating MSCs migration potential.For in vivo experiment,diabetic wound healing mice model was generated to elucidate the effect of Sa-pretreated MSCs transplantation in wound closure rate,as well as re-epithelization status,observed with hematoxylin and eosin staining.The diabetic wound healing mice model were divided into three groups:1)mice injected with PBS,2)mice transplanted with PBS-pretreated MSCs,and 3)mice transplanted with Sa-pretreated MSCs.Results(1)Hyperglycemic condition induced the generation of ROS and suppressed total cell number of MSCs,while Sa treatment into MSCs restored these hyperglycemia-induced alterations.In line with this,total apoptotic cells were also suppressed by treating MSCs with Sa.The expression level of cell survival factor,heme-oxygenase 1(HO-1),was enhanced in Sa-pretreated MSCs.Further treatment of HO-1 inhibitor into Sa-pretreated MSCs nullified the ROS level and total apoptotic cells,indica-ting the importance of HO-1 in mediating the Sa-induced survival of MSCs under hyperglycemia.(2)Transwell chamber and scratch assay results showed that Sa-pretreated MSCs have a higher migration potential under hyperglycemia,supported by higher F-actin polymerization fractal dimension.Fibroblast growth factor 2(FGF2)and hepatocyte growth factor(HGF)expression level,which are essential factors for cell migration,were also improved in Sa-pretreated MSCs under hyperglycemia.(3)In diabetic wound healing mice model,transplantation of Sa-pretreated MSCs resulted in significantly improved wound closure rate and re-epithelization.The protein levels of HO-1,FGF2,and HGF were also enhanced in the tissues obtained from the wound site of diabetic wound healing mice model which were transplanted with Sa-pretreated MSCs.Conclusions Salidroside pretreatment on MSCs could improve their survival and migration potentials,subsequently promoting wound healing process under hyperglycemia.This prospective MSC-based therapy could serve as a novel strategy to improve diabetic wound healing.
文摘Cancer stem cells(CSCs)are the driving force for sustainable tumor growth and metastasis and responsible for drug resistance and cancer relapse.Nanoparticle-based drug delivery has been demonstrated to be effective in combating tumor growth.However,it has been challenging to selectively eliminate CSCs due to the lack of a general signature for a spectrum of cancers.It is known that CSCs from various types of cancer show lower stiffness compared to non-CSCs.It remains unclear whether low stiffness in CSCs influences cellular uptake in nanoparticle-based drug delivery and thus the chemotherapy efficacy.Graphene quantum dot(GQD)is emerging as a promising carrier material in delivering anti-cancer drugs.We found that breast CSCs were softer than conventional cancer cells,which were further softer compared to healthy breast tissue cells.Importantly,soft CSCs uptook more GQD than conventional cancer cells,while stiff breast cancer cells with relatively low stiffness uptook more GQD than healthy breast cells.Softening cells by pharmacologically inhibiting actomyosin activity using either siRNA or actomyosin inhibitors significantly enhanced the cellular uptake of GQD in breast cancer cells but not CSCs,while stiffening cells by activating actomyosin using CA-MLCK/ROCK or actomyosin activators considerably suppressed the nanoparticle uptake in both cancer cells and CSCs.GQD could specifically target CSC because of low cell stiffness of CSC in breast cancer cell line MCF-7 and MDA-MB-231.Further regulating cell stiffness reflected that decreasing breast cancer cell stiffness by inhibiting actomyosin activity using blebbistatin could promote GQD uptake.Vice versa,stiffening cancer cell by activating actomyosin decreased GQD uptake.The attachment of anti-cancer drug doxorubicin did not alter the trend of GQD uptake in neither soften nor stiffen cancer cells.Actomyosin activity regulates cellular uptake ofGQD might through clathrin and caveolin-mediated endocytosis.Cancer cells are softer than normal cells from the same organ,CSC are softer than non-CSC.Thus we further confirmed that the GQD uptake of normal breast cell line MCF-10 is less than breast cancer cell line MCF-7 and MDA-MB-231.Suggesting the clinical significance that using GQD as drug carrier targeting softer cells could reduce side effects to normal tissue cells.Since CSC are softer than non-CSC,GQD could decreased the percentage of CSC in whole cancer cell population by targeting softer cells.These results suggesting that it would be possible to target cancer cells and CSC by targeting from a perspective of cell mechanical difference.High uptake of nanoparticles in soft cancer cells could not be explained by their differential membrane potentials.Mechanistically,low cell mechanics or inhibiting actomyosin activity activated both clathrin and caveolin-mediated endocytosis signaling pathways,while high cell mechanics or activating actomyosin suppressed these signalings.Pharmacologically inhibiting clathrin or caveolin-mediated endocytosis signaling significantly decreased GQD uptake in CSCs and in conventional breast cancer cells when actomyosin was suppressed.Further,GQD conjugated with doxorubicin could be specifically delivered into CSCs with low stiffness and eliminated more CSCs in the presence of both CSCs and non-CSCs.Taken together,these data reveal the regulatory role of cell mechanics in cellular uptake of nanoparticles and demonstrate that GQD can be utilized to specifically eliminate CSCs,which have important implications in nanoparticle-based drug delivery for cancer therapy.
基金National Key Research and Development Program(2016YFC1306301).
文摘OBJECTIVE To establish an in vitro cel model based on patient-specific human microglia to study the pathological mechanism of Alzheimer disease(AD) and to screen candidate drugs.METHODS First,the induced pluripotent stem cells(iPSCs) of AD patients and cognitive normal controls(CNC) were induced to hematopoietic progenitor cells(HPCs),and then HPCs were further induced with IL-34,M-CSF,GM-CSF and TGF-β1 for 20 d to obtain microglialike cells(MGLCs).HPCs were isolated by flow cytometry and MGLCs were identified by immunofluorescence.Cell phagocytosis was determined by phagocytosis neutral red experiusing Luminex assay kits,and the cell growth curve during the experiment was recorded by IncuCyte ZOOM.The phagocytic ability and secretion of cytokines of MGLCs were observed under the stimulation of LPS.RESULTS MGLCs from AD patients(AD-MGLCs) and CNC expressed microglia markers IBA1,TMEM119,P2 RY12,TREM2 and CD11 B.The results of phagocytosis neutral red experiment showed that under normal conditions,AD-MGLCs had stronger phagocytic ability(P<0.01).Stimulation by LPS resulted in increased phagocytosis of cel s,and the increase in phagocytosis of CNC-MGLCs was higher than AD-MGLCs(P<0.01).Experiments showed that high concentrations of LPS(>2 mg·L^(-1)) resulted in CNC-microglia death(P<0.01),whereas ADMGLCs did not show significant death.The cytokine assay showed that under normal conditions,the concentrations of IFN-γ and IL-2 secreted by AD patients were slightly higher than those of CNC.After LPS stimulation,the secretion of TNF-α,IL-6 and IL-10 was significantly increased.The increased secretion of AD-MGLCs was greater than that CNC-MGLCs(P<0.01).CONCLUSION AD-iPSCs derived MGLCs exhibit significant inflammatory characteristics and are more active than CNC,which may be associated with chronic inflammatory responses caused by microglia in AD,thus may provide valuable new tools to screen candidate drugs for the disease and to discover the mechanisms underlying AD pathogenesis.
基金National Natural Science Foundation of China(8157381381173598)+1 种基金Excellent Talent Program of Chengdu University of Traditional Chinese Medicine(YXRC2019002)Fund of Scientific Research Innovation Team Construction in Sichuan Provincial University(18TD0017)
文摘OBJECTIVE To investigate the inhibitory effect of scutellarin on the self-renewal and differentiation of HT-29 cells-derived cancer stem-like cells(HT-29CSC)in vitro and in vivo,and to explore its mechanism.METHODS The effect of scutellarin on the growth of HT-29CSC was determined by 3D Culture assay.The effect of scutellarin on growth and transformation of HT-29CSC was probed by soft agar colony formation assay.The effect of scutellarin on the differentiation of HT-29CSC was determined by serum induction differentiation assay in vitro.The effects of scutellarin on the expressions of marker gene Lgr5,target gene c-Myc,proliferation gene CK20 and Nanog gene were measured by quantitative real-time RT-PCR.Investigate the effect of scutellarin on the expression of c-Myc,Gli1,and Lgr5 protein by Western blotting.A subcutaneous xenograft model of colon cancer in nude mice was established and administered by intraperitoneal injection.The change of body weight and tumor size of nude mice were observed every two days.Investi⁃gate the effects of scutellarin on the growth of xenograft tumors in nude mice.The expression of CD133,Lgr5,Gli1,Ptch1,c-Myc,Ki67,CK20,Nanog gene in tumors were measured by quantitative real-time RT-PCR.The expression of c-Myc,Gli1,Lgr5,CD133,Ki67 protein were measured by Western blotting.RESULTS Scutellarin can inhibit the growth of HT-29CSC in 3D culture.Compared with the solvent control group,scutellarin can significantly inhibit the growth and transformation and differentiation of HT-29CSC in vitro(P<0.01).The expression levels of marker genes Lgr5,target gene c-Myc,proliferation gene CK20 and Nanog in HT-29CSC were down-regulated by scutellarin.Scutellarin can reduce the expression of c-Myc,Gli1,and Lgr5 protein in HT-29CSC.Scutellarin can inhibit the growth of colon cancer xenografts,lower CD133,Lgr5,Gli1,Ptch1,c-Myc,Ki67,CK20,and Nanog mRNA level of xenograft tumors,reduce the expression of c-Myc,Gli1,Lgr5,CD133,and Ki67 protein of xenograft tumors in nude mice.CONCLUSION Scutellarin,which is the main component of scutellaria barbata,can inhibit the differentiation of HT-29CSC and the mechanism is to inhibit the activity of Hedgehog signaling pathway.
基金Supported by the Scientifi c Research Foundation for Doctors of Northeast Agricultural University(2012RCB27)Open Projects of Key Laboratory of Animal Genetics,Breeding and Reproduction,College of Heilongjiang Province(GXZDSYS-2012-07)
文摘The experiment was designed to study effects of retinoic acid and ascorbic acid on differentiation of mouse embryonic stem cells to cardiomyocytes. Embryonic bodies （EB） were developed from mESC in suspension culture, different levels of concentration of retinoic acid and ascorbic acid were used to determine the optimal conditions for EB formation. The results showed that the optimal concentrations were 10.9 mol. L-1 and 0.1 mg. mL-1 for retinoic acid and ascorbic acids, respectively. 50% of EB which was significantly （p〈0.05） different from the control group developed to cardiomyocytes. In conclusion, rctinoic acid and ascorbic acid had strong ability to promote cardiomyocyte differentiation of mouse embryonic stem cells. 10-9 mol. L-1 retinoic acid and 0.10 mg. mL-1 ascorbic acids were recommended to induce differentiation of mouse ES ceUs toward cardiomyocytes.
文摘Non-alcoholic fatty liver disease(NAFLD)is the most common chronic liver disease,defined by several phases,ranging from benign fat accumulation to non-alcoholic steatohepatitis(NASH),which can lead to liver cancer and cirrhosis.Although NAFLD is a disease of disordered metabolism,it also involves several immune cell-mediated inflammatory processes,either promoting and/or suppressing hepatocyte inflammation through the secretion of pro-inflammatory and/or anti-inflammatory factors to influence the NAFLD process.However,the underlying disease mechanism and the role of immune cells in NAFLD are still under investigation,leaving many open-ended questions.In this review,we presented the recent concepts about the interplay of immune cells in the onset and pathogenesis of NAFLD.We also highlighted the specific non-immune cells exhibiting immunological properties of therapeutic significance in NAFLD.We hope that this review will help guide the development of future NAFLD therapeutics.
文摘Objective The binding of cell adhesive peptides(such as RGD)to integrins initiates the recruitment of cytoplasmic adaptor proteins(e.g.,vinculin)and the formation of focal adhesion(FA)complexes required for cell adhesion.The ability to manipulate this ligand-mediated cell adhesion process is crucial for regulating cell migration,cell differentiation,injury healing,and immune response.Some recent studies reported the importance of the tether length/mobility of the cell adhesive ligands in regulating the traction force development of cells.In the native cellular microenvironment,such a dynamic change in the nanoscale tether length of bioactive ligands is often mediated by conformational changes of the structural proteins due to protein folding or degradation.However,no prior studies have demonstrated the modulation of the ligand tether mobility by controlling the intramolecular folding of polymeric linkers.Unfoldable synthetic macromolecules with easy synthetic routes and controllable structures,such as supramolecular host-guest single chain nanogels(SCNGs),are ideal candidates for mimicking the changes in the tether mobility of bioactive ligands via biorthogonal triggers.Methods S,S’-bis(a’a’-dimethyl-a’’-propargyl acetate)trithiocarbonate was first used to mediate the RAFT polymerization of N,N-dimethyl acrylamide,vinyl-adamantane and vinyl-β-cyclodextrin to yield the ADA@CD-SCNGs.The preparation of the unfoldable host-guest SCNGs was evidenced by the by gel permeation chromatography,proton nuclear magnetic resonance spectroscopy,atomic force microscopy and dynamic light scattering.Then the RGD peptide was conjugated to the alkynyl group on one end of the SCNGs before immobilizing the material on the substrate,which was confirmed by scanning electron microscopy(SEM).The regulation of cell behaviours by unfolding of the SCNG-RGD was confirmed by immunofluorescence staining of vinculin and Yes-associated protein(YAP).Results The preparation of ADA@CD-SCNGs was confirmed by GPC which showed a unimodal molecular weight distribution.DLS and AFM data also proved that the SCNGs had an average diameter of 12±3nm.SEM images showed that SCNGs were conjugated as a linker of RGD peptide to thiolated glass substrate at an average density of 162±11 particles/μm2.These particles disappeared after adding free competitive ADA guest molecules,indicating the triggered unfolding of the tether SCNGs.In addition,the unfolding of supramolecular ADA@CD-SCNGs was also evidenced by a decrease in the GPC elution time and a slight increase in the apparent molecular weight.These results show that the immobilized ADA@CD-SCNGs can be unfolded to tune the tether length and mobility of the conjugated RGD ligands.Then we investigated the regulation of the cell behaviors on the substrate by triggering the unfolding of SCNG linkers.A critical level of traction force is required to effectively initiate and maintain integrin-mediated formation of FA complexes and subsequent mechano-transduction signaling.An increased tether length in cell-adhesive ligands can lead to a diminished cell traction force as if cells are adhering to soft substrates.Here,the unfolding of the ADA@CD-SCNG-RGD triggered by the addition of free ADA led to disassembly of the mature focal adhesions in the cells as evidenced by the reduced vinculin and F-actin in staining.Subsequently,nuclear YAP also decreased significantly because of the impaired mechano-sensing and diminished cell cytoskeleton tension.In addition,the extensively spread cells gradually became round after the medium was supplemented with free competitive ADA to unfold the SCNG linker.These finding demonstrates that the substrates with the unfolded ADA@CD-SCNG-RGD only supported weak cell adhesions.In contrast,on the substrate conjugated with the nonunfoldable MBA-SCNG-RGD linker,the addition of free ADA resulted in no change in the spread cell morphology and protein expressions.These results indicate that the unfoldable host-guest ADA@CD-SCNG can be used to manipulate the nanoscale presentation of ligands to regulate cell behaviors.Conclusions We demonstrate the application of SCNGs as the supramolecular linker to tune the nanoscale ligand tether length.These findings demonstrate that the strategy of manipulating the tether mobility of bioactive ligands by using supramolecular SCNGs as linkers provides a highly tunable,biomimetic,and bio-orthogonal approach to study the dynamic events of cell adhesion.
文摘Aim The present study aims to investigate whether BMSCs transplantation may inhibit hypertrophic hearts and its underlying mechanisms. Background There is no evidence so far that Bone marrow-derived mesenchy- mal stem cells （BMSCs） can heal pathological myocardial hypertrophy. Methods To observe the antihypertrophic actions, BMSCs was indirectly cocultured with NRVCs in vitro, or intramyocardially transplanted into hypertrophic hearts in vivo. Results ISO-induced typical hypertrophic characteristics of cardiomyocytes were obviously preven- ted by BMSCs in the co-culture model in vitro and after BMSCs transplantation in vivo. Furthermore, the activation of the Ca2＋/calcineurin/NFATc3 hypertrophic pathway was shown abrogated in the presence of BMSCs both in vitro and in vivo. Interestingly, blockage of VEGF release from BMSCs but not bFGF and IGF-1 can abolish the protec- tive effects of BMSCs on cardiomyocytes hypertrophy. Consistently, VEGF administration attenuated ISO-induced BNP and β-MHC expression and the activation of Ca2＋/cal- the enlargement of cellular size, the augment of ANP, cineurin/NFATc3 hypertrophic pathway, and these can be abrogated by blocking VEGFR-1, indicating VEGFR-1 is involved in the antihypertrophic role of VEGF. We further find that the ample VEGF secretion contributing to the anti-hypertrophic effects of BMSCs originates from BMSCs interplay with cardiac cells but not BMSCs or cardiomyo- cytes alone. Conclusions Thus, mesenchymal stem cells are able to inhibit myocardial hypertrophy via interacting with cardiomyocytes so as to promote VEGF release which inhibits the activation of the Ca2＋/calcineurin/NFATc3 hypertrophic signaling pathway in cardiac cells, in addition to its well-recognized ability to ameliorate myocardial injuries by replacing dead cells.
文摘Embryonic stem (ES) cell biology is attracting much attention in cell biology because of their pluripotent behaviors and potential therapeutic applications. However,what maintains ES cell pluripotency and what triggers ES cell
基金supported by funds from Huazhong University of Science and Technology,Wuhan,China
文摘Spinal cord injury repair is one of the major challenges in medicine,as it can lead to permanent loss of function of central nervous system and damage to other function of the body.Stem cell transplantation together with tissue engineering is increasingly becoming a potential choice of treatment.However,direct transplantation of stem cells without scaffolds has yielded poor clinical outcome.Here we show a strategy of using mouse embryonic stem cells(ESCs)cultured within a silk fibroin(SF)based,three-dimensional scaffold with oriented channels by a directional temperature field freezing technique and lysophilization.We find that the ESCs maintained proliferation and migrated in the scaffolds and the cells migrated fastest along the SF channels.SF scaffolds contributed to ESC differentiation into neural and glial cell like cells and expressions of the neural and glial cell markers MAP2 and GFAP were greatly elevated when retinoic acid was used as an inducing factor.Our results suggest that this approach may offer some hope in the future for spinal cord injury repair using SF scaffolds and ESCs.
基金Natural Science Foundation of Hebei Province(H2022106065)Scientific Research Program of Hebei Provincial Administration of Traditional Chinese Medicine(2023172)。
文摘OBJECTIVE To explore the key mechanism of Bazi Bushen capsule(BZBS)in delaying the senescence of mesenchymal stem cells(MSCs)through network pharmacology and in vitro experiments.METHODS Network pharmacology was used to predict the mechanism targets of BZBS in delaying MSCs senescence.A MSCs senescence model induced by D-galactose(D-gal)was used to investigate the effect and mechanism of BZBS on MSCs senescence in vitro.RESULTS Network pharmacology analy⁃sis showed that BZSB could delay MSCs senes⁃cence.The experiment showed that BZBS could significantly improve the survival activity of the aged MSCs.It significantly reduced the positive rate ofβ-galactosidase staining and p16,p21 expression in aged MSCs,enhanced the ability of adipogenic differentiation and osteogenic differentiation,and increased expression of Nanog,OCT4 and SOX2 in senescent MSCs.CONCLU⁃SIONS Network pharmacology and in vitro cell experiments verified that BZBS could delay MSCs senescence.
基金This study was partially supported by a grant from the ministry of HealthL abor and Welfare of Japan+1 种基金Hum an Genom e and Regenerative Medicine Project (ChairpersonHidehiko Saito)
文摘Objective Bone-marrow stem-cell transplantation has been shown to improve cardiac function in patients with AMI, but the safety of intracoronory infusion of autologous peripheral blood stem-cell(PBSCs) in patients with AMI is unknown. For this reason, we observe the feasibility and safety of PBSCs transplantation by intracoronory infusion in such patients.Method Fourty one patients with AMI were allocated to receive Granulocyte Colony-Stimulating Factor (G-CSF:Filgrastim,300 μg) with the dose of 300 μg-600 μg/day to mobilize the stem cell, and the duration of applying G-CSF was 5 days . On the sixth day, PBSCs were separated by Baxter CS 3000 blood cell separator into suspend liquid 57 ml. Then the suspend liquid was infused into the infarct related artery (IRA)by occluding the over the wire balloon and infusing artery through balloon center lumen. In the process of the intracoronary infusion of PBSCs, the complications should be observed, which were arrhythmias including of bradycardia, sinus arrest or atrial ventricular block, premature ventricular beats ,ventricular tachycardia, ventricular fibrillation; and hypotention, etc. Results There were total 10 cases with complications during the intracoronary infusion of PBSCs. The incidence of complications was 24.4%(10/41), including bradycardia is 2.4 %(1/41), sinus arrest or atrial ventricular block is 4.9%(2/41), ventricular fibrillation is 2.4 %( 1/41), hypotention is14.6 % (6 /41).Conclusions In patients with AMI, intracoronary infusion of PBSCs is feasible and safe.
文摘Aim Aging is an independent risk factor for heart disease, however the effective intervention has not been found so far. Bone marrow mesenchymal stem cells （BMSCs） have been shown to offer a wide variety of cel- lular functions including the protective effects on damaged hearts. Here we investigated the antiaging properties of BMSCs and the underlying mechanism in a cellular model of cardiomyocyte senescence and a rat model of aging hearts. Methods In vitro study, neonatal rat ventricular cells （NRVCs） and BMSCs were cocultured in the same dish with a semipermeable membrane to separate the two populations. In vivo, the BMSCs were injected into the rat hearts to observe their antiaging effects. The expression of β-galactosidase and aging-related proteins, and the lev- els of oxidative stress were determined in vivo and in vitro. The heart function was measured by the High-Resolution Imaging System. Results Monocultured NRVCs displayed the senescence-associated phenotypes, characterized by an increase in the number of β-galaetosidase-positive cells and decreases in the degradation and disappearance of cellular organelles in a time-dependent manner. The levels of reactive oxygen species and malondialdehyde were el- evated, whereas the activities of antioxidant enzymes superoxide dismutase and glutathione peroxidase were de- creased, along with upregulation of p53, p21cipl/wafl and p16INK4a in the aging eardiomyoeytes. These deleterious alterations were abrogated in aging NRVCs cocultured with BMSCs. Qualitatively, the same senescent phenotypes were consistently observed in aging rat hearts. Notably, BMSC transplantation significantly prevented these detri- mental alterations and improved the impaired cardiac function in the aging rats. Conclusions BMSCs possess strong antiseneseence action on the aging NRVCs and hearts and can improve cardiac function after transplantation in aging rats. The present study, therefore, provides an alternative approach for the treatment of heart failure in the elderly population.
基金supported by National key research and development program(2016YFC1306300)
文摘OBJECTIVE To establish an in vitro cell model based on patient-specific human neural stem cells to study the pathomechanism of sporadic AD as well as screen candidate drugs.METHODS The peripheral blood cells from sporadic AD patients and cognitive normal controls were repro.grammed into inducedpluripotent stem cells(iPSCs),which were further induced into neural stem cells and neurons.The cell growth curve during the differentiation process was recorded by the IncuCyte ZOOM,and neural stem cells and neurons were identified by immunofluorescence.The apoptosis of neural stem cells and neurons was detected by Click-iT~Plus TUNEL Assay.RESULTS Neural stem cells derived from AD patients and cognitive normal controls can express neural stem cell markers Nes.tin,Sox1,Sox2 and Ki67.TUNEL assay results showed that the number of TUNEL-positive cells in neu.ral stem cells derived from AD patients was significantly higher than that of cognitive normal controls(P<0.01).When neural stem cells were differentiated into neurons,the percentage of MAP2 positive cells in the neural stem cell-derived culture dish of AD patients was significantly higher than the cogni.tive normal controls at day 16 of neuronal differentiation(P<0.01);the TUNEL assay showed that the number of TUNEL-positive cells in AD-derived neurons was significantly greater than that in cognitive normal controls(P<0.01) at day 16 of neuronal differentiation.CONCLUSION Our study revealed that AD-iPSC-derived neural stem cells exhibit premature neuronal differentiation and increased neural apoptosis,which might be relevant to the neuronal loss of AD,thus may provide valuable new tools to screen candidate drugs for the disease and to discover the mechanisms underlying AD pathogenesis.
