摘要
目的 了解脐血造血干细胞在体外扩增培养过程中的生长分化特点 ,探讨脐血干细胞与体外扩增后的祖细胞混合移植的可行性以及合适的移植时机。方法 接种脐血单个核细胞(MNC)于含体积浓度为 2 0 %胎牛血清 (FCS)的IMDM悬浮培养体系中 ,加入干细胞因子 (SCF)、Flt 3配基 (FL)、粒 单细胞集落刺激因子 (GM CSF)、白细胞介素 3(IL 3)、白细胞介素 6 (IL 6 ) ,培养 2 1d ,每周半量换液 ,同时进行细胞计数 ,涂片形态学观察 ,粒 单系祖细胞 (CFU GM )、红系祖细胞 (BFU E)集落培养 ,细胞表面CD34 + 、CD33+ 、CD3+ 、CD19+ 标记和细胞增殖周期的动态观察。结果 培养7d时的脐血细胞 ,CFU GM、BFU E集落形成最多 ,之后逐渐减少 ,2 1d时已低于培养前水平。CD34 + 细胞比例在培养后 7d达高峰 ,14d时开始下降 ;CD33+ 细胞比例在培养后 7～ 14d达高峰 ,2 1d时开始下降 ;CD3+ 、CD19+ 细胞比例均在培养第 7d时显著下降 ,并持续低水平至 2 1d。培养 7d后G0 期细胞比例明显减少 ,S期细胞快速增高 ,至 2 1d时仍有较高比例。培养第 2 1d的细胞涂片见少量中、晚幼粒细胞。结论 理论上脐血干细胞与扩增后的祖细胞混合移植用于成人具有可行性 ,体外扩增培养
ObjectivesTo explore the growth and differentation of umbilical cord blood (UCB) hematopoietic cells in vitro expansion and investigate the probability of mix transplantation with UCB hematopoietic stem cells (HSC) and expanded hematopoietic progenitor cells (HPC) in vitro and the optimal time for mix transplantation. MethodsMononuclear cells from UCB were cultured for 21 days in IMDM mediums containing 20% FCS and SCF, FL, GM CSF, IL 3, IL 6. Cultivating mediums were half exchanged every week. The total cells counting, smear morphology, CFU GM, BFU E colonys, cell cycle and cellular markers were analyzed dynamically every week. ResultsTotal cells were increased 387.8 fold at day 7 and went on increasing in later days, but the increasing tendency was slower. CFU GM, BFU E colonies reached the highest level at day 7 (P< 0.01), then deceased, and kept lower level at day 21 than that precultured (P< 0.01). CD34 + cells reached the highest proportion at day 7 (P< 0.01), decreased at day 14 and decreased further at day 21 (P< 0.01); CD33 + cells reached the highest proportionat day 7 14 (P< 0.01), and decreased at day 21 (P< 0.01); Both CD3 + cells and CD19 + cells were notably decreased at day 7 (P< 0.01), and kept lower proportion till day 21. The cells in the S phase were increased quickly at day 7 and kept high level till day 21. A few myelocytes and metamyelocytes were observed on the cell smear at day 21. Conclusions HPC can largely expanded and directly differentiated while UCB hematopoietic cells were in vitro expansion. The proportion of T and B lymphocyte was decreased after expansion. It is possible that the mix transplantation with HPC and expanded HPC would be used on adults. Culturing 7 days is the optimal time for mix transplantation.
出处
《中华器官移植杂志》
CAS
CSCD
北大核心
2001年第6期336-338,共3页
Chinese Journal of Organ Transplantation
