论文部分内容阅读
目的研究可溶性CD40L(sCD40L)对人脐血CD34+细胞体外扩增和集落形成的影响。方法用磁珠阳性分离纯化法分离脐血CD34+细胞;用造血干细胞体外悬浮培养、集落形成实验以及流式细胞仪检测等方法,观察应用sCD40L对脐血造血干细胞扩增、CD34+细胞增殖及造血集落形成的影响。结果sCD40L能显著增强干细胞因子(SCF)、白细胞介素3(IL-3)、促红细胞生成素(EPO)、粒细胞-巨噬细胞集落刺激因子(GM-CSF)对脐血干细胞总数和CD34+细胞的扩增作用,并能促进造血集落,尤其是粒细胞-巨噬细胞集落形成单位(CFU-GM)的形成。sCD40L联合其他造血细胞生长因子对脐血CD34+细胞的扩增作用在培养的第7d最显著,当sCD40L浓度为40μg/L时,CD34+细胞的扩增倍数达8.23±1.26。结论sCD40L与SCF、IL-3、EPO、GM-CSF联合应用,对人脐血造血干细胞的体外扩增更为有效。
Objective To study the effect of soluble CD40L (sCD40L) on the proliferation and colony formation of human umbilical cord blood CD34 + cells in vitro. Methods The cord blood CD34 + cells were isolated by magnetic beads positive isolation and purification method. The effects of sCD40L on cord blood hematopoietic stem cell proliferation, CD34 + cell proliferation and hematopoietic colonies were observed by suspension culture, colony formation assay and flow cytometry in vitro. The formation of the impact. Results sCD40L could significantly enhance the effects of SCF, EPO and GM-CSF on the total number of cord blood stem cells and CD34 + Cell expansion and promote the formation of hematopoietic colonies, especially granulocyte-macrophage colony forming units (CFU-GMs). The effect of sCD40L combined with other hematopoietic growth factors on the proliferation of cord blood CD34 + cells was the most significant at day 7 of culture. When the concentration of sCD40L was 40μg / L, the multiplication of CD34 + cells was 8.23 ± 1.26. Conclusion The combined application of sCD40L with SCF, IL-3, EPO and GM-CSF is more effective in vitro expansion of human umbilical cord blood hematopoietic stem cells.