论文部分内容阅读
目的 :评价CD34 +细胞体外扩增时高增殖潜能集落形成细胞 (HPP CFC)的变化。方法 :利用mini MACS分离脐血CD34 +细胞 ,流式细胞仪 (FACS)测定细胞纯度 ( 86%～ 95% )。应用含rhSCF、rhIL 3、rhIL 6和rhGM CSF的培养体系扩增CD34 +细胞。分别于培养的第 1,2和 4周收取细胞进行HHP CFC测定 ,测试体系为含重组人SCF、GM CSF、IL 3、IL 6、EPO等细胞因子的半固体甲基纤维素。结果 :测试体系中加入单个细胞因子不能产生HPP CFC ,含rhIL 3和rhGM CSF体系产生的集落小且主要为巨噬细胞集落 ,由所有因子组成的体系产生的集落大而致密 ,包含粒系 3种类型。此外 ,尽管细胞总数显著增加 ,扩增的CD34 +细胞HPP CFC形成率随培养时间的延长而减少。结论 :本方法显示 ,CD34 +细胞体外扩增中可以保持HPP CFC数量。
Objective: To evaluate the changes of high proliferative potential colony forming cells (HPP CFCs) in vitro when CD34 + cells were expanded. Methods: The umbilical cord blood CD34 + cells were isolated using mini MACS and the cell purity (86% -95%) was determined by flow cytometry (FACS). CD34 + cells were expanded using a culture system containing rhSCF, rhIL 3, rhIL 6 and rhGM CSF. The cells were harvested at 1, 2, and 4 weeks of culture for HHP CFC measurement. The test system was semi-solid methylcellulose containing recombinant human SCF, GM CSF, IL 3, IL 6, EPO and other cytokines. Results: The addition of a single cytokine to the test system failed to produce HPP CFCs. Colonies produced by the rhIL 3-containing and rhGM-CSF-derived colonies were small and predominately macrophage colonies. Colonies produced by all factors produced large and dense colonies containing the granulocyte 3 Kind of type In addition, although the total number of cells increased significantly, the rate of HPC CFC formation in the expanded CD34 + cells decreased with increasing incubation time. Conclusion: This method shows that the number of HPC CFCs can be maintained during the in vitro expansion of CD34 + cells.