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
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 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
Objective:To investigate the influence of electroacupuncture(EA) combined with repetitive transcranial magnetic stimulation(rTMS) on the temporal profile of nestin expression after induction of focal cerebral ischemia...Objective:To investigate the influence of electroacupuncture(EA) combined with repetitive transcranial magnetic stimulation(rTMS) on the temporal profile of nestin expression after induction of focal cerebral ischemia in adult rats and to explore the mechanism of EA combined with rTMS in treating ischemic brain injury.Method:The model of transient focal ischemia was produced by occlusion of middle cerebral artery.Seventy-five Wistar rats were randomly divided into normal group,model group,EA group,rTMS group and EA +rTMS group.The neurologic impairment rating and ability of learning and memory were observed at the 7th、14th and 28th d after infarction respectively.Meanwhile,Western blotting was used to observe the number of nestin expression positive cells.Result:Nestin-positive cells were found in cortex,subgranular zone(SGZ),subventricular zone(SVZ) of the ipsilateral side at different time points after cerebral ischemia.The number of nestin-positive cells peaked at the 7th d,began to decrease at the 14th d and was significantly higher in EA+rTMS group than that in model group(P< 0.05),then almost reached normal at the 28th d.The improvement of neural motor function deficits as well as the indexes of learning and memory were more obvious in EA+rTMS group compared with model group(P<0.01,P<0.05).These effects were most obvious in EA +rTMS group compared with the EA and rTMS group(P<0.05).Conclusion:EA and rTMS possess the potency of building up and can increase the number of nestin-positive cells in some brain regions after focal cerebral ischemia,which might be one of the important mechanisms of EA combined with rTMS in treating ischemia brain injury.展开更多
为探讨切割穹窿海马伞海马提取液和银杏叶提取物(extract of ginkgo bilobo,EGb)在海马NSCs向胆碱能神经元定向分化中的作用,分别制备大鼠穹窿海马伞切割侧和正常侧海马提取液;将从鼠胚海马中分离扩增的NSCs球分成4组,应用不同的培养液...为探讨切割穹窿海马伞海马提取液和银杏叶提取物(extract of ginkgo bilobo,EGb)在海马NSCs向胆碱能神经元定向分化中的作用,分别制备大鼠穹窿海马伞切割侧和正常侧海马提取液;将从鼠胚海马中分离扩增的NSCs球分成4组,应用不同的培养液促其分化:(1)联合组:含切割侧海马提取液和银杏内酯的DMEM/F12培养基;(2)提取液组:含切割侧海马提取液的DMEM/F12培养基;(3)EGb组:含银杏内酯的DMEM/F12培养基;(4)对照组:含正常侧海马提取液的DMEM/F12培养基。培养14d后行ChAT免疫荧光检测,计算ChAT阳性神经元的分化率,图像处理细胞面积和周长。结果显示联合组各项指标均明显优于其它各组(P<0.01);提取液组、EGb组各项指标也均优于对照组(P<0.05);细胞面积提取液组优于EGb组(P<0.05),细胞周长EGb组优于提取液组(P<0.05),两组ChAT阳性神经元分化率无明显差异(P>0.05)。上述提示切割穹窿海马伞的海马提取液和EGb联合应用可诱导海马NSCs分化为更多、更为成熟的胆碱能神经元。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 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.
基金Supported by the National Nature Science Foundation of China(No.30672216)Project of Wuhan Hygiene Bureau(No.WX08A01,No.WZ08B02)
文摘Objective:To investigate the influence of electroacupuncture(EA) combined with repetitive transcranial magnetic stimulation(rTMS) on the temporal profile of nestin expression after induction of focal cerebral ischemia in adult rats and to explore the mechanism of EA combined with rTMS in treating ischemic brain injury.Method:The model of transient focal ischemia was produced by occlusion of middle cerebral artery.Seventy-five Wistar rats were randomly divided into normal group,model group,EA group,rTMS group and EA +rTMS group.The neurologic impairment rating and ability of learning and memory were observed at the 7th、14th and 28th d after infarction respectively.Meanwhile,Western blotting was used to observe the number of nestin expression positive cells.Result:Nestin-positive cells were found in cortex,subgranular zone(SGZ),subventricular zone(SVZ) of the ipsilateral side at different time points after cerebral ischemia.The number of nestin-positive cells peaked at the 7th d,began to decrease at the 14th d and was significantly higher in EA+rTMS group than that in model group(P< 0.05),then almost reached normal at the 28th d.The improvement of neural motor function deficits as well as the indexes of learning and memory were more obvious in EA+rTMS group compared with model group(P<0.01,P<0.05).These effects were most obvious in EA +rTMS group compared with the EA and rTMS group(P<0.05).Conclusion:EA and rTMS possess the potency of building up and can increase the number of nestin-positive cells in some brain regions after focal cerebral ischemia,which might be one of the important mechanisms of EA combined with rTMS in treating ischemia brain injury.
文摘为探讨切割穹窿海马伞海马提取液和银杏叶提取物(extract of ginkgo bilobo,EGb)在海马NSCs向胆碱能神经元定向分化中的作用,分别制备大鼠穹窿海马伞切割侧和正常侧海马提取液;将从鼠胚海马中分离扩增的NSCs球分成4组,应用不同的培养液促其分化:(1)联合组:含切割侧海马提取液和银杏内酯的DMEM/F12培养基;(2)提取液组:含切割侧海马提取液的DMEM/F12培养基;(3)EGb组:含银杏内酯的DMEM/F12培养基;(4)对照组:含正常侧海马提取液的DMEM/F12培养基。培养14d后行ChAT免疫荧光检测,计算ChAT阳性神经元的分化率,图像处理细胞面积和周长。结果显示联合组各项指标均明显优于其它各组(P<0.01);提取液组、EGb组各项指标也均优于对照组(P<0.05);细胞面积提取液组优于EGb组(P<0.05),细胞周长EGb组优于提取液组(P<0.05),两组ChAT阳性神经元分化率无明显差异(P>0.05)。上述提示切割穹窿海马伞的海马提取液和EGb联合应用可诱导海马NSCs分化为更多、更为成熟的胆碱能神经元。
