探讨CB CD34^ 细胞扩增的最佳HGF组合及移植时机。方法：CB CD34^ 细胞培养于无血清培养液中，分别加入下列HGF，A组：对照组（无HGF）；B组：SCF，IL－3，IL－6；C组：SCF，IL－3，IL－6，G－CSF，Epo;D组：SCF，IL－3，IL－6，TPO，Flt3-L；E组：SCF，IL－3，IL－6，TPO，Flt3-L,G-CSF,Epo。培养22d，对不同培养时间的细胞进行NC数量及系特异性CD动态观察。结果：同对照组相比，扩增组NC和CD34^ 细胞均有不同程度的扩增。NC和CD34^ 细胞扩增高峰分别在第10天和第6天。E组的扩增效果最佳。CD检测显示，各组CD34^ 及CD34^ CD^38-细胞的比例逐渐减少，但B，D2组的CD34^ CD38^-细胞的比例在第6天有一过性回升，CD154^ ,CD13^ ,CD61^ 及Gly-A^ 细胞的比例则逐渐升高，D组CD34^ 及CD34^ ,CD38^-细胞的比例明显地高于E组（P＜0．01），CD3^ 和CD19^ 细胞的比例在扩增中呈下降趋势，结论：HSPC分化程度与HGF组合及培养时间有关。就细胞总数而言，E组HGF组合最佳，但就HSPC含量来说，以D组为优，第1周扩增产物中HSPC含量较高，移植时间以扩增的第6－10天为宜。

To explore the optimal combination of HGF for expanding CD34 cells and the suitable time for transplantation.Methods: CB CD34 cells were cultured for 22 days in medium containing following combination of HGFs: Group A (Containing no HGFs as control), group B (SCF,IL-3,IL-6), group C (SCF,IL-3,IL-6,G-CSF,Epo),group D (SCF,IL-3,IL-6,TPO,Flt3-L), group E (SCF,IL-3,IL-6,TPO,Flt3-L,G-CSF,Epo). At every intervals, the cells were collected and assayed for CD. Results: NC and CD34 cells were expanded significantly in all expansion group, the peak of number of NC is on day 10 and that of CD34 cells is on day 6. The highest level of NC is in group E, and that of CD34 and CD34 CD38 - cells is in group D. The content of CD34 and CD34 CD38 - cells decreased gradually. Interestingly, the content of CD34 CD38 - cells in group B and D increased slightly on day 6 and that of CD14 ,CD13 ,CD61 Gly-A cells elevated increasingly correspondingly. However, the content of CD3 and CD19 cells decreased during expansion. Conclusions: Differentiation of CD34 cells related to combination of HGF and time of ex vivo expansion , SCF combines with IL-3,IL-6,TPO,Flt3-L,G-CSF and Epo is optimal combination regarding to the number of NC, but in view of the content of HSPC, the combination containing SCF,IL-3,IL-6,TPO and Flt3-L is suitable for expansion of HSPC. The content of HSPC reached the highest level in first week, so the suitable time for transplantation of expanded cells is from day 6 to day 10 of expansion.

国家"九五"医学科技攻关课题基金(编号96-906-01-17)国家自然科学基金(批准号3987077)及上海血液学研究所胡应洲基金部分资助