不同细胞因子对脐血CD34+细胞扩增的影响

目的探讨不同细胞因子促进脐血CD34 细胞扩增的效应,为下一步的临床应用奠定基础。方法采用EasySep免疫磁珠阳性选择系统分离脐血中CD34 细胞,在无血清、无基质细胞培养体系中添加不同细胞因子培养2周,分组:A组,SCF FL TPOB组,SCF FL TPO IL-3C组,SCF FL TPO G-CSF。结果CD34 细胞数目于培养后7d达到峰值,3组间差异无显著性;B组扩增至14d时细胞总数达(384.40±17.48)倍,显著高于另外2组。扩增7d后的脐血细胞经体外培养可形成造血集落。结论体外扩增脐血CD34 细胞理想的细胞因子组合为SCF TPO FL,以7d为宜。

Effect of different cytokines on the proliferation of CD34+ hematopoietic cells in cord blood

Objective] To investigate the proliferative effect of different cytokines on the CD34 hematopoietic cells in umbilical cord blood. Methods] In this study, a serum-free, stroma-free medium for hematopoietic stem cells proliferation was systematically developed. CD34 cells isolated with the EasySep human CD34 positive selection system from umbilical cord blood were cultured in vitro with different cytokines for 14 days to evaluate proliferative potential. Group A: SCF FL TPO, group B: SCF FL TPO IL-3, group C: SCF FL TPO G-CSF. Results] The number of CD34 cells reached peak at the 7th day with no significant difference among 3 groups. While addition of the growth factors SCF, TPO, FL and IL-3 to serum-free media did allow high proliferation of hematopoietic cells with the most total nuclear cells after 7 and 14 days of culture. CFU-G and CFU-GEMM formed after plantation of 7 d-proliferated cord blood hematopoietic cells into methylcellulose media. Conclusions] These data indicate that SCF, FL and TPO can be used to efficiently proliferate hematopoietic progenitors from human umbilical cord blood.