【摘要】 目的:探讨造血生长因子不同组合方式对脐血CD34+造血干/祖细胞(HSC/HPC)的扩增作用。方法:应用StemsepTM阴性分选系统分离纯化CD34+细胞,在体外液体培养体系中经不同组合细胞因子进行扩增1￣3w。结果:FL+Tpo+SCF+IL-3+GM-CSF+G-CSF+Epo组合的扩增效果最佳,可分别扩增细胞总数、造血祖细胞集落总数(CFCs)、CD34+细胞达1627.57±52.93、56.78±8.13、41.09±4.11倍。FL和Tpo具有明显的协同扩增早期祖细胞的作用,CFCs和CD34+细胞在1￣2w维持在较高水平,以后逐渐衰退。结论:合理的细胞因子组合在大量扩增造血细胞的同时也有助于早期造血祖细胞的自我更新,把握适宜的扩增时机,将有助于扩增细胞在质和量上满足移植和基因治疗等的要求。更多还原

【Abstract】 Objective: To investigate the role of recombinant human hematopoietic growth factors(HGFs) in expansion of purified CD34+ cells derived from human cord blood.Methods: A sterilized separation system for purification of CD34+ hematopoietic stem / progentior cells(HSC/HPC) was established by means of StemsepTM according to the strategy of negative selection on immunomagnetic separation.CD34+ cells were expanded with various combination of cytokines in a 21-day liquid culture systems.Results: A combination of seven HGFs including FL,Tpo,SCF,IL-3,GM-CSF,G-CSF and Epo was identified as the most potent combination of those tested,and it increased total nucleated cell by 1627.57±52.93 fold,total progenitor cells(CFCs) by 56.78±8.13 fold and CD34+ cell by 41.09±4.11 fold,respectively.Among the HGFs examined,FL and Tpo were the most potent ones for expanding cord blood CD34+ cells.Maximal expansions of CFCs and CD34+ cells were observed during the first two weeks of culture.Conclusion: Under conditions of appropriate HGFs combination and expansion duration,it is possible to expand cord blood CD34+ cells ex vivo for transplantation and therapeutic gene transfer.更多还原