摘要
【目的】探讨人骨髓间充质干细胞(MSC)及其培养上清、融冻产物对脐血CD34+细胞扩增及分化的作用。【方法】分离纯化人骨髓MSC,收集纯化三代的MSC培养上清,采用反复冻融法获得MSC融冻产物,采用免疫磁珠分选系统纯化脐血CD34+细胞。实验分为4组,分别为含MSC滋养层组、MSC上清组、MSC融冻产物组及单纯造血生长因子(HGFs)对照组。流式细胞仪检测人脐血CD34+细胞在这四种培养体系中第6天和第12天有核细胞数及CD34+、CD34+CD38-、CD41+、CD19+、CD3+细胞含量。【结果】①MSC滋养层组对脐血有核细胞和对脐血CD34+、CD34+CD38-的扩增能力最强,MSC培养上清组和MSC滋养层相比具有类似的作用,但其作用强度减弱(P<0.05)。②MSC培养上清和MSC均具有抑制脐血CD34+细胞分化为CD3+和CD19+细胞的作用,差异无显著性(P>0.05)。③MSC培养上清和MSC均具有促进脐血CD34+细胞向CD41+细胞分化的作用,MSC滋养层的作用强于MSC培养上清(P<0.05)。④MSC融冻产物已完全丧失对脐血CD34+细胞扩增和分化作用,与单纯因子对照组相似(P>0.05)。【结论】MSC培养上清对脐血CD34+、CD34+CD38-细胞均具有扩增作用,而且可促进脐血CD34+细胞向CD41+细胞分化。
【Objective】 To investigate the effects of mesenchymal stem cells (MSC) feeder layer,culture sera and freeze- thaw lysates on expansion and differentiation of cord blood CD34+ cells in vitro.【Methods】 MSC were isolated from human bone marrow and cultured until the third passage.Sera were obtained from the cultured MSC,and freeze-thaw lysates were obtained by repeated freeze-thaw procedures.Cord blood CD34+ cells were isolated by magnetic cell separation system,and were co-cultured with the MSC feeder layer,culture sera,freeze-thaw lysates and hematopoietic growth factors (HGFs),respectively.The nucleated cells,CD34+ cells,CD34+ CD38- cells,CD41+ cells and CD3+ cells in the above culture system were detected by flow cytometry on day 6 and day 12.【Results】 ①MSC feeder layer had a strong effect on nucleated cells,CD34+,CD34+ CD38- cells expansion.The MSC sera and freeze-thaw lysates had similar effect on cell expansion,but the effect was weaker than that of feeder layer (P 0.05).②Both MSC sera and feeder layer inhibited cord blood CD34+ cells differentiation toward CD3+ cells or CD19+ cells,and no significant differences were found between these two groups (P 0.05).③Both MSC sera and feeder layer promoted cord blood CD34+ cells differentiation toward CD41+ cells,and the effect was stronger in the feeder layer than that of the sera (P 0.05).④Freeze-thaw lysates had no effect on cell expansion and differentiation,and were similar with that of HGFs (P 0.05).【Conclusions】 The MSC sera have positive effects on expansion of cord blood CD34+ and CD34+ CD38- cells,moreover they have the ability of promoting cord blood CD34+ cells differentiation toward CD41+ cells.
出处
《中山大学学报(医学科学版)》
CAS
CSCD
北大核心
2010年第1期39-44,共6页
Journal of Sun Yat-Sen University:Medical Sciences
基金
国家自然科学基金(30872786)
广东省科技计划引导项目(2008B030301305)