论文部分内容阅读
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 细胞扩增
Objective : To investigate the in vitro expansion of cord blood hematopoietic stem/progenitor cells. METHODS: Immunomagnetic beads were used to separate and purify umbilical cord blood AC133+ cells and were expanded in vitro under the action of cytokine FL T3 ligand and thrombopoietin to detect the expansion of nucleated cells. Flow cytometry was used to analyze changes in cell surface markers. RESULTS : FL T3 and thrombopoietin were cultured in vitro for 2 weeks. The nucleated cells were expanded (18 ± 8) fold. CD3 4 cells were amplified 2. 8-fold, AC133+ was not amplified, the purity of CD3 4 cells decreased from ( 56 ± 2 3) % to (8 ± 1) %, and AC133 + cells decreased from 85% to ( 0.1 ± 1. 0 .1) %. As the in vitro culture time was extended to 4 weeks, nucleated cells and CD+3 4 were further amplified. Amplified 475 times and 17 times, respectively. However, the cells differentiated, and the proportion of CD+3 4 cells in the nucleus cells dropped to 2%, and AC133+ cells disappeared. CD45 cells rose to 1.3%. Conclusion : FL and TPO long-term culture of AC133+ cells can expand CD3 4 cells