以人胎肝基质细胞为饲养层有效扩增脐带血CD34~＋细胞

目的以人胎肝基质细胞（fetal liver stromal cell,FLSC）作为饲养层,进行脐带血CD34＋细胞的扩增培养,寻找更加有效的体外扩增方法。方法比较脐带血CD34＋细胞在无饲养层和人FLSC饲养层条件下,培养7,13,15d的细胞增殖状况;并利用流式细胞仪检测不同时间点CD34＋细胞的比例。结果随着培养时间的延长,脐带血CD34＋细胞的扩增呈现时间依赖性改变;在FLSC饲养层条件下,CD34＋细胞培养7,13,15d的扩增数量分别为（11.83±0.76）×105,（14.37±0.32）×105和（18.73±0.25）×105,而在无饲养层条件下的扩增数量分别为（2.90±0.10）×105,（8.87±0.15）×105和（11.97±0.15）×105,两者相比有显著性差异（P（0.01）;此外,流式细胞仪检测各时间点CD34＋细胞的比例,在培养第13天,FLSC饲养层条件下CD34＋细胞的比例为10.21%,而在无饲养层为1.01%,两者存在显著差异。结论以人FLSC作为饲养层可以有效扩增脐带血造血干/祖细胞,为今后临床造血干细胞移植提供充足的细胞来源。

In vitro expansion of umbilical cord blood CD34+ cells over human fetal liver stromal cells

Objective To seek novel in vitro expansion methods to expand umbilical cord blood CD34＋ cell over human fetal liver stromal cells（FLSC）.Methods Expansion results of umbilical cord blood CD34＋ cells were evaluated by cell count and flow cytometry analysis and compared between using FLSC-feeder cells co-culture and feeder-free culture systems.Results After 7d,13d,15d culture,the expansion of CD34＋ cells using FLSC-feeder cells co-culture system was significantly better than that using the feeder-free culture system,with CD34＋ cell count being（11.83±0.76）×105,（14.37±0.32）×105 and（18.73±0.25）×105 using FLSC-feeder cells co-culture system,respectively,compared with（2.90±0.10）×105,（8.87±0.15）×105,and（11.97±0.15）105 using the feeder-free culture system.After 13d culture,analysis of flow cytometry data showed significant difference in CD34＋ cell percentages between using FLSC-feeder cells co-culture system and using the feeder-free culture system（10.21% versus 1.01%）.Conclusion Umbilical cord blood hematopoietic stem/progenitor cells can be efficiently expanded when FLSC-feeder cells co-culture system is used,which may provide sufficient hematopoietic stem/progenitor cells for clinical transplantation.