FL联合TPO体外培养AC133^+细胞表面标记的变化

THE CHANGE OF SURFACE MARKERS OF AC133^+ CELLS CULTURED WITH FL AND TPO IN VITRO

目的 :探讨脐血造血干 /祖细胞的体外扩增。方法 :免疫磁珠法分离纯化脐血 AC133+细胞 ,在细胞因子 FL T3配体、血小板生成素作用下 ,体外扩增 ,检测有核细胞扩增的倍数。采用流式细胞仪分析细胞表面标志的变化。结果 :FL T3联合血小板生成素体外培养 2周。有核细胞扩增 ( 18± 8)倍。CD3 4 细胞扩增 2 .8倍 ,AC133+未获扩增 ,CD3 4 细胞纯度由 ( 5 6± 2 3) %下降到 ( 8± 1) % ,AC133+细胞由 85 %下降到 ( 0 .1± 0 .1) %。随着体外培养时间延长至 4周 ,有核细胞 ,CD+3 4 进一步扩增。分别扩增 475倍和 17倍。但细胞随之发生分化 ,CD+3 4 细胞占有核细胞中的比例下降至 2 % ,AC133+细胞消失。 CD45 细胞上升到 1.3%。结论 :FL联合 TPO长期培养 AC133+细胞能使CD3 4

Objective:To explore the expansion of progenitor cells from human umbilical cord blood in vitro . Methods:Cord blood AC133 + cells was isolated by using MACS,expanded with FLT3 ligand and TPO in vitro.The expansion folds of nuclear cells were detected and the change of cells surface markers was analyzed with flow cytometry.Results:After 2 weeks culture,the number of nuclear cells increased approximately 18±8 folds,and CD + 34 cells about 2.8 folds.AC133 + cells haven't been expanded.The percentage of CD + 34 has dropped from (56±23)% to (8±1)% and AC133 +cells from 85% to (0.1± 0.1)%.After 4 weeks,the number of nuclear cells and CD + 34 cells has been increased more about 475 folds and 17 folds.But nuclear cells turn to differentiation from then on. The percentage of CD + 34 cells has dropped to 2%.AC133 + cells has disappeared,CD + 45 cells has increase up to 1.3%.Conclusion:This result showed that AC133 + cells cultured with FLT3 and TPO can be used to expand CD + 34 cells in vitro.