条件培养液对脐血CD_(34)^+细胞扩增效应的实验研究

EXPERIMENTAL STUDY ON EXPANSION OF HUMAN UMBILICAL CORD BLOOD CD_(34)^+ CELLS IN VARIOUS CONDITIONED MEDIA

目的探讨并分析不同条件培养液对脐血CD+ 34 细胞的体外扩增效应及影响因素。方法利用 4种条件培养液 (胎儿肌肉、胎盘组织、外周血、脐血单个核细胞 )对脐血CD+ 34 细胞进行 2周的体外扩增 ,并和三细胞因子组合 :干细胞因子 (stemcellfactor ,SCF)、血小板生成素 (thrombopoietin ,TPO)、flt 3配基 (flt 3ligand ,FL) ,对脐血CD+ 3 4 细胞扩增结果进行了比较 ;利用ELISA方法检测 4种条件培养液中细胞因子 [白细胞介素 1(interleukin 1,IL 1)、白细胞介素 6 (interleukin 6 ,IL 6 )、FL、TPO]的浓度。结果脐血单个核细胞条件培养液及细胞因子组合对脐血CD+ 3 4 细胞均有明显的扩增效应 ,脐血组优于细胞因子组合 ;4种条件培养液中 ,脐血组细胞因子FL的含量明显高于其余条件培养液组。结论脐血条件培养液可作为体外培养扩增脐血CD+ 34 细胞有效而廉价的扩增剂 ;细胞因子FL可能是造成不同条件培养液对脐血CD+ 34

ObjectiveTo investigate the ability of different conditioned media for expansing ex vivo umbilical cord blood(UCB) CD + 34 cells and to analyse possible reasons. MethodsPurified CD + 34 cells of UCB were expansed using various conditioned media (up to 2 weeks). The results were compared with those of three cytokine group (stem cell factor+thrombopoietin+flt 3 ligand)and the concentration of cytokines in conditioned media was examined using the method of ELISA.ResultsThe conditioned medium of UCB nucleared cells and three cytokine group showed efficient expansion of purified UCB CD + 34 cells and the content of flt 3 ligand(FL) in UCB conditioned medium were significantly higher than the other groups.ConclusionThe UCB conditioned medium can be used for ex vivo effective expansion UCB CD + 34 cells, and FL might play an important part in the process of ex vivo expansion UCB CD + 34 cells.