不同细胞因子对脐血CD_(34)^+细胞扩增的影响

Effect of different cytokines on the proliferation of CD_(34)^+ hematopoietic cells in cord blood*

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。

[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.