【摘要】 目的　探讨脐血造血细胞体外扩增及其移植的最佳时机。方法　用粒细胞集落刺激因子、粒 巨噬细胞集落刺激因子、重组人白细胞介素 3、重组人白细胞介素 6、干细胞因子和红细胞生成素长期培养脐血造血细胞 ,并对其表面标志及细胞增殖周期进行动态分析。结果　脐血中CD3 + 细胞、CD4 + /CD4 5RO+ 和CD8+ /CD4 5R0 + 细胞培养 3d稍有增加 ,培养 7d达高峰 ,14d显著下降 ,培养 2 1d时处于更低水平。CD4 + 和CD8+ 细胞培养 3～ 7d没有明显变化 ,培养 14d显著下降 ,培养 2 1d下降更加明显。CD4 + /CD4 5RA+ 和CD8+ /CD4 5RA+ 细胞培养 3d后开始下降 ,14d后明显下降。HLA DR+ 细胞在培养中变化不大 ,只在培养 3～ 7d稍有增加。CD3 4+ 细胞在培养 3d开始增多 ,培养 7d时进入高峰 ,培养 14d时开始下降 ;培养 7d后CD3 4+ 、CD3 4+ CD3 8-、CD3 4+ CD3 8+ 细胞约为培养前的 8.8,7.4,8.7倍 [培养前CD3 4+ 、CD3 4+ CD3 8-、CD3 4+ CD3 8+ 细胞的测定值分别为 (3.46± 1.2 1) %、(0 .45± 0 .34 ) %和 (3.2 6±1 0 9) % ;培养 7d后 ,测定值分别为 (30 .6 0± 6 .0 3) % ,(3.40± 3.32 ) %和 (2 8.33± 14.84) % ]。细胞增殖周期分析显示 ,培养 3d后G0 期细胞比例迅速减少 ,而S期细胞比例快速增加 ,到培养 2 1d时 ,S期更多还原

【Abstract】 Objective To explore the optimal timing for in vitro expansion and transplantation of umbilical cord blood hematopoietic cells. Methods Hematopoietic cells from human umbilical cord blood were cultured with G CSF 、GM CSF、rIL 3、rIL 6、SCF and Epo in a long term culture system, the cell cycle and cellular markers were analysed dynamically. Results CD 3 +、CD 4 +/CD 45RO + and CD 8 +/CD 45RO + cells increased at day 3, and reached the highest level at day 7, decreased at day 14,and remained lower level at day 21. CD 4 + and CD 8 + cells showed no change at day 7, decreased at day 14, and decreased further at day 21. CD 4 +/CD 45RA + and CD 8 +/CD 45RA + cells decreased after day 3, and decreased further at day 14. HLA DR + cells increased from day 3 to day 7. CD 34 + cells increased at day 3, reached the highest level at day 7, and decreased at day 14. CD 34 +、CD 34 + CD 38 -、CD 34 + CD 38 + cells increased 8.8,7.4 and 8.7 fold at day 7, respectively, as compared with that precultured. The cells in the S phase increased at day 21. Conclusions This culture system is able to expand hematopoietic cells of human umbilical cord blood and keeps them intact.The number of hematopoietic cells reached the highest level in the culture from day 7 to day 14, which is the optimal time for transplantation.更多还原