骨髓基质细胞共培养体系促进人脐血CD133^+细胞的体外扩增  

Coculture system containing marrow stromal cell layer supporting in vitro expansion of umbilical cord blood CD133+ cells

作　　者：郝思国[1] 孙关林[1] 邬维礼[1] 吴英理[1] 

机构地区：[1]上海第二医科大学附属瑞金医院上海血液学研究所,200025

出　　处：《中华器官移植杂志》2005年第1期28-32,共5页Chinese Journal of Organ Transplantation

基　　金：国家自然科学基金(30270570);上海市博士点建设基金;上海血液学研究所胡应州基金部分资助

摘　　要：目的探讨来自白血病患者骨髓细胞的基质细胞层建立的扩增体系体外扩增人脐带血造血干/祖细胞的可行性。方法应用白血病患者缓解期骨髓基质细胞,结合造血细胞生长因子体外扩增人脐带血CD133+细胞(实验组),对扩增细胞的免疫表型、集落形成能力等生物学特性进行动态观察,并与单独应用造血细胞生长因子的常规扩增(对照组)进行比较。结果基质细胞层上接种的细胞易于收集;实验组在扩增各时间点的有核细胞数(NC)明显高于对照组,而且有核细胞中CD34+细胞、CD133+细胞、CD34+CD38-细胞以及CD34+CD133+细胞的比例也明显高于对照组;实验组扩增各时间点有核细胞各种集落的种植率明显高于对照组,有核细胞、CD34+细胞、CD133+细胞、CD34+CD38-细胞以及细胞集落的扩增倍数明显高于对照组。结论含有基质细胞的扩增培养体系,不仅可以加速人脐带血造血干/祖细胞的体外扩增,而且可以延缓造血干/祖细胞的分化。Objective To investigate the effects of expansion system containing bone marrow stromal cell layer on proliferation and differentiation of umbilical cord blood umbilical cord blood (UCB) haematopoietic stem/progenitor cells (HSPC) ex-vivo expansion and define the optimal means of expansion of UCB HSPC.Methods UCB CD133+ cells were cultured in expansion system containing bone marrow stromal cell layer established using bone marrow from patients with APL in CR and CML in CP. The number of NC, colonogeneic assay, immuno-phenotyping of the NC at every interval were monitored. Results The expanded cells could be collected easily in the culture containing marrow stromal cells. The number and the expansion folds of NC from stromal culture were significantly more than those from control. The number of CD34+, CD133+, CD34+CD38-cells and ratio of CD133+/CD34+ in stromal culture were remarkably higher than those in controls. The expansion of ~above subsets in NC from stromal culture was remarkably more than that from controls. Meanwhile, the plate efficiencies and expansion folds of the colonies of NC from stromal culture were also ~remarkably more than those from controls. Conclusion This expansion system could promote the ~expansion of UCB HSPC and also delay the differentiation of expanded cells, which could remain the primitive HPC in the expanded products and was helpful for engraftment of expanded cells.

关 键 词：造血干细胞 骨髓细胞 细胞培养 胎血 白血病 

分 类 号：R733.7[医药卫生—肿瘤]