CD34+ cells from human umbilical cord blood were purified by Dynal beads M-450 CD34 immunoselection system and cultured in the presence of various cytokines alone or in combination, including stem cell factor (SCF), i...CD34+ cells from human umbilical cord blood were purified by Dynal beads M-450 CD34 immunoselection system and cultured in the presence of various cytokines alone or in combination, including stem cell factor (SCF), interleukin-6 (IL-6) and erythropoietin (EPO). The results revealed that: (D In methylcellulose culture, the plating efficiencies of purified cord blood CD34+ cells were much different when stimulated by various cytokines. IL-6 alone had the lowest colo-ny yield, while the combination of SCF, IL-6 and EPO had the highest yield. ② In the suspension culture, IL-6 alone or IL-6 + EPO had little expanding effect on cord blood CD34+ celis, the other cytokine combinations could expand cord blood CD34+ celis at different Ievels. Among them, the combination of SCF, IL-6 and EPO had the maximal expanding effect on cord blood CD34+ celis, the number of progenitor celis peaked at day 21, about 29-fold increase and nucleated celis increased approximately 3676-fold at day 28. The expanding effect of展开更多
To induce the growth and differentiation of dendritic cells (DCs) from human cord blood, CD34 + cells isolated from human cord blood by mini MACS were cultured in a liquid culture system with rhSCF, rhGM CSF, rhTNF α...To induce the growth and differentiation of dendritic cells (DCs) from human cord blood, CD34 + cells isolated from human cord blood by mini MACS were cultured in a liquid culture system with rhSCF, rhGM CSF, rhTNF α and rhFL for 10 days. Then the induced cells were characterized by DC′s morphological and phenotypic properties. In addition, they stimulated the proliferation of allogeneic T cells and possessed an efficient capacity for initiating T cell dependent antitumor immune responses in vitro. It is concluded that mature DCs could be obtained from human cord blood CD34 + cells.展开更多
本研究旨在观察几种不同细胞因子组合通过体外培养以诱导造血干/祖细胞增殖和向巨核细胞/血小板分化。应用无血清培养基(S tem Span(SFEM)体外扩增脐血CD34+细胞并向巨核细胞/血小板定向分化,将不同细胞因子组合分为3个阶段培养,并比较...本研究旨在观察几种不同细胞因子组合通过体外培养以诱导造血干/祖细胞增殖和向巨核细胞/血小板分化。应用无血清培养基(S tem Span(SFEM)体外扩增脐血CD34+细胞并向巨核细胞/血小板定向分化,将不同细胞因子组合分为3个阶段培养,并比较其培养效果。结果表明,在第一阶段的第14天时,SCF+TPO+FL+IL-3组细胞扩增倍数最高为11 000±1 000,显著高于SCF+TPO+FL组,但与SCF+TPO+IL-3组及SCF+TPO+FL+IL-3+羟皮质激素组相比较无显著差异;在第二培养阶段的第7天时,SCF+TPO+FL+IL-11组巨核细胞系扩增倍数为204666.7±11718.9,显著高于SCF+TPO+FL+IL-3组,与SCF+TPO+FL+IL1-1+BM P4+VEGF组比较并无显著差异。在第三阶段中,SCF+TPO+FL+IL-11和SCF+TPO+FL+IL-11+BM P4+VEGF2组的CD41+血小板样细胞所占比例显著高于SCF+TPO+FL+IL-3组。结论:SCF+TPO+FL+IL-3因子组合可显著提高造血干/祖细胞的扩增倍数,SCF+TPO+FL+IL-11组合有利于巨核细胞的扩增成熟及分化,为下一步的体外培养巨核细胞/血小板体系的建立奠定了一定的基础。展开更多
本研究探讨高迁移率族蛋白B1(high mobility group box1,HMGB1)对人脐血CD34+细胞迁移的影响及其机制。用流式细胞仪检测脐血CD34+细胞表面HMGB1受体:晚期糖基化终产物受体(receptor for advancedglycation end products,RAGE)、Toll样...本研究探讨高迁移率族蛋白B1(high mobility group box1,HMGB1)对人脐血CD34+细胞迁移的影响及其机制。用流式细胞仪检测脐血CD34+细胞表面HMGB1受体:晚期糖基化终产物受体(receptor for advancedglycation end products,RAGE)、Toll样受体2(Toll-like receptors2,TLR2)及TLR4的表达。用磁珠分选法富集的新鲜人脐血CD34+细胞,经不同浓度的HMGB1(10、50、100、1000ng/ml)刺激后,应用transwell小室趋化装置观察HMGB1对人脐血CD34+细胞的迁移活性,显微镜下计数,计算趋化指数,即试验孔与对照孔细胞数的比值。以未加HMGB1的培养细胞为对照组。结果表明:人脐血CD34+细胞经免疫磁珠分选富集的纯度达98%以上。HMGB1在一定的浓度范围内随浓度递增对人脐血CD34+细胞的迁移作用逐渐增强,当HMGB1浓度为100ng/ml时迁移活性最强,与对照组比较差异显著(p<0.01)。抗-RAGE抗体可部分抑制HMGB1对脐血CD34+细胞的迁移作用。结论:一定浓度的HMGB1加强人脐血CD34+细胞迁移功能,此作用有可能通过RAGE介导。展开更多
采用两步法分离出脐血CD34^+细胞,比较研究了混合脐血血浆联合IL-3,IL-6,GM-CSF,Epo 4种中、晚期造血因子和单纯的4种造血因子情况下脐血CD34^+细胞的体外扩增。结果表明,混合脐血血浆联合造血因子对粒-巨噬细胞集落形成单位(granulocyt...采用两步法分离出脐血CD34^+细胞,比较研究了混合脐血血浆联合IL-3,IL-6,GM-CSF,Epo 4种中、晚期造血因子和单纯的4种造血因子情况下脐血CD34^+细胞的体外扩增。结果表明,混合脐血血浆联合造血因子对粒-巨噬细胞集落形成单位(granulocyte-macrophage colony-forming unit,CFU-GM),红系爆式集落形成单位(burst-forming unit of erythriod,BFU-E),混合集落形成单位(minxed colony-forming unit,CFU-mix)3种集落的扩增效果明显优于单纯的4种造血因子联合组,差异具有统计学意义(P<0.01),但单纯混合脐血血浆扩增效果较差。脐血造血细胞扩增对子成人脐血移植有重要意义。上述结果提示,混合脐血血浆的扩增成功,可代替或弥补早期造血生长因子的作用,用于脐血造血细胞的体外扩增。展开更多
文摘CD34+ cells from human umbilical cord blood were purified by Dynal beads M-450 CD34 immunoselection system and cultured in the presence of various cytokines alone or in combination, including stem cell factor (SCF), interleukin-6 (IL-6) and erythropoietin (EPO). The results revealed that: (D In methylcellulose culture, the plating efficiencies of purified cord blood CD34+ cells were much different when stimulated by various cytokines. IL-6 alone had the lowest colo-ny yield, while the combination of SCF, IL-6 and EPO had the highest yield. ② In the suspension culture, IL-6 alone or IL-6 + EPO had little expanding effect on cord blood CD34+ celis, the other cytokine combinations could expand cord blood CD34+ celis at different Ievels. Among them, the combination of SCF, IL-6 and EPO had the maximal expanding effect on cord blood CD34+ celis, the number of progenitor celis peaked at day 21, about 29-fold increase and nucleated celis increased approximately 3676-fold at day 28. The expanding effect of
文摘To induce the growth and differentiation of dendritic cells (DCs) from human cord blood, CD34 + cells isolated from human cord blood by mini MACS were cultured in a liquid culture system with rhSCF, rhGM CSF, rhTNF α and rhFL for 10 days. Then the induced cells were characterized by DC′s morphological and phenotypic properties. In addition, they stimulated the proliferation of allogeneic T cells and possessed an efficient capacity for initiating T cell dependent antitumor immune responses in vitro. It is concluded that mature DCs could be obtained from human cord blood CD34 + cells.
文摘本研究探讨高迁移率族蛋白B1(high mobility group box1,HMGB1)对人脐血CD34+细胞迁移的影响及其机制。用流式细胞仪检测脐血CD34+细胞表面HMGB1受体:晚期糖基化终产物受体(receptor for advancedglycation end products,RAGE)、Toll样受体2(Toll-like receptors2,TLR2)及TLR4的表达。用磁珠分选法富集的新鲜人脐血CD34+细胞,经不同浓度的HMGB1(10、50、100、1000ng/ml)刺激后,应用transwell小室趋化装置观察HMGB1对人脐血CD34+细胞的迁移活性,显微镜下计数,计算趋化指数,即试验孔与对照孔细胞数的比值。以未加HMGB1的培养细胞为对照组。结果表明:人脐血CD34+细胞经免疫磁珠分选富集的纯度达98%以上。HMGB1在一定的浓度范围内随浓度递增对人脐血CD34+细胞的迁移作用逐渐增强,当HMGB1浓度为100ng/ml时迁移活性最强,与对照组比较差异显著(p<0.01)。抗-RAGE抗体可部分抑制HMGB1对脐血CD34+细胞的迁移作用。结论:一定浓度的HMGB1加强人脐血CD34+细胞迁移功能,此作用有可能通过RAGE介导。
文摘采用两步法分离出脐血CD34^+细胞,比较研究了混合脐血血浆联合IL-3,IL-6,GM-CSF,Epo 4种中、晚期造血因子和单纯的4种造血因子情况下脐血CD34^+细胞的体外扩增。结果表明,混合脐血血浆联合造血因子对粒-巨噬细胞集落形成单位(granulocyte-macrophage colony-forming unit,CFU-GM),红系爆式集落形成单位(burst-forming unit of erythriod,BFU-E),混合集落形成单位(minxed colony-forming unit,CFU-mix)3种集落的扩增效果明显优于单纯的4种造血因子联合组,差异具有统计学意义(P<0.01),但单纯混合脐血血浆扩增效果较差。脐血造血细胞扩增对子成人脐血移植有重要意义。上述结果提示,混合脐血血浆的扩增成功,可代替或弥补早期造血生长因子的作用,用于脐血造血细胞的体外扩增。
