间充质干细胞条件培养基和细胞因子联合应用扩增造血干细胞

Combined Application of Mesenchymal Stem Cell Conditioned Medium and Cytokines to Expand Hematopoietic Stem Cells

目的:优化造血干细胞(hematopoietic stem cells,HSCs)体外扩增方案,提高扩增效率。方法:从脐带血中通过免疫磁珠分选法获得CD34+HSCs;组织块贴壁法原代分离培养获得脐带间充质干细胞(mesenchymal stem cells,MSCs),扩增后收集条件培养基;将固定细胞因子组合与MSCs条件培养基联合应用于HSCs扩增培养,通过细胞计数、流式检测及集落分析来判定扩增效应。结果:联合培养方案的造血细胞的数量、CD34+HSCs的比例及集落形成的数量均明显多于单独利用细胞因子组合时。结论:MSCs条件培养基和细胞因子联合应用能明显提高HSCs扩增效率。

Objective To optimize the hematopoietic stem cell(HSC)in vitro expansion protocol to improve expansion efficiency.Methods CD34+HSCs were obtained from umbilical cord blood by immunomagnetic bead separation.The umbilical cord mesenchymal stem cells(MSCs)were obtained by primary isolation and culture of tissue block adherence method,and conditioned medium was collected after expansion.MSC conditioned medium combined with fixed cytokines were used in the expansion and culture of HSCs,and the expansion effect was determined by cell counting,flow cytometry and colony analysis.Results The number of hematopoietic cells,the proportion of CD34+HSCs and the number of colony formation in the combination culture scheme were significantly higher than those in the cytokine treatment alone.Conclusion The combination of MSCs conditioned medium and cytokines can significantly improve the amplification efficiency of HSCs.