脐血CD34^+CD38^-早期造血祖细胞的体外增殖和凋亡

Proliferation and Apoptosis in vitro of Umbilical Cord Blood CD34^+ CD38 ^- Hematopoietic Early Progenitor Cells

为了从适龄产妇脐血中分离出CD34+CD38-细胞,在体外较长时间培养后观察分析CD34+CD38-细胞分 裂增殖、凋亡以及干细胞因子对CD34+CD38-细胞增殖的影响,用流式细胞仪从10例健康产妇脐血中分选出 CD34+和CD38-标记的脐血原始细胞,在添加IL-3、IL-6、GM-CSF、EPO、IGF-1和SCF 6种混合因子的干细胞培养基 中培养6个月,观察细胞并绘制生长曲线,用单细胞凝胶电泳检测干细胞因子对CD34+CD38-细胞生长的影响和 用流式细胞仪检测CD34+ CD38-细胞凋亡情况。结果表明:脐血CD34+CD38-细胞可在体外较长时间培养增殖, 无异常或过度的细胞凋亡发生。结论:通过控制培养条件,脐血CD34+CD38-早期造血祖细胞可在体外较长时间 培养增殖,以作为大量脐血原始细胞移植的细胞来源。

To cultivate CD34 + CD38 cells isolated from umbilical cord blood of healthy puerperal women over a longer-period of time for observation of cell division, proliferation, apoptosis, and effects of stem cell factor on the growth of CD34 + CD38 - cells, with flow cytometry, CD34 + CD38 - cells were isolated from umbilical cord blood of 10 healthy puerperal women and cultivated in stem cell media with supplement of IL-3, IL-6, GM-CSF, EPO, IGF-1 and SCF 6 kinds cell growth stimulating factors for six months. The cell growth curves were established. The effects of stem cell factor on the growth of CD34 + CD38- cells and cell apoptosis were investigated with the single cell gel electrophoresis technique and flow cytometry method, respectively. The results showed that CD34 + CD38 - cells isolated from umbilical cord blood were capable of proliferating after being cultivated in vitro over a longer-period of time with no evidence of the presence of excessive apoptosis. In conclusion, under appropriate culture conditions, CD34 + CD38 - hematopoietic early progenitor cells from umbilical cord blood can serve as a resource providing a large amount of primitive cells for transplantation therapy after a longer period of cultivation and proliferation in vitro.