脐血单个核细胞和CD34^+细胞体外扩增造血干/祖细胞

Hematopoietic Stem/Progenitor Cell Expansion from Umbilical Cord Blood CD34^+ Selected Cells and MNC

考察了脐血单个核细胞 ( MNC)和 CD3 4 +富集细胞在 SCF+IL- 3 +IL- 6 +FL+Tpo细胞因子组合下的体外扩增特性。实验发现 :CD3 4 +富集细胞具有很高的扩增潜力 ,通过 3周培养总细胞扩增了 ( 883 .2± 3 2 2 .4)倍 ,而 MNC仅扩增了 ( 5 3 .3± 6 .2 )倍。对比细胞集落生成和 CD3 4 +细胞的扩增发现 ,MNC的集落密度由最初的 ( 2 70 .9± 5 4 .9) CFCs/ 1 0 5cells上升至第 7天的( 1 496 .3± 41 7.7) CFCs/ 1 0 5cells,CD3 4 +细胞百分比则由最初的 ( 0 .99± 0 .1 8) %上升至第 7天的( 4.4± 1 .9) % ,而 CD3 4 +富集细胞在培养过程中 ,虽然集落总数和 CD3 4 +细胞总数始终呈上升趋势 ,但是其集落密度和 CD3 4 + 细胞百分比则始终呈下降趋势。在两种起始细胞培养中 ,CFU- GM(粒细胞 -巨噬细胞集落形成单位 )在集落中的含量逐渐增加 ,BFU- E(爆式红细胞集落形成单位 )含量则逐渐降低。在 3周的短期培养中 ,MNC可以得到更多的集落形成细胞 ( CFC)和 CD3 4

The expansion characteristics of mononuclear cells(MNC) and CD34 + selected cells were examined by culturing the two kinds of cells in the medium containing cytokine cocktails of SCF+IL-3+IL-6+FL+Tpo. It was found that the CD34 + selected cells presented a high proliferation potential. Within 3 weeks of culture period, the fold expansion of total cells in CD34 + selected cell cultures achieved 883.2±322.4, while that of MNC reached to 53.3±6.2 only. In the cultures of MNC cells, from day 0 to day 7, the colony density increased from (270.9±54.9)/10 5 cells to (1 496.3±417.7)/10 5 cells, and the proportion of CD34 + cells increased from (0.99±0.18)% to (4.4±1.9)%. However, in the cultures of CD34 + selected cells, the total number of CFCs(colony-forming cell) and CD34 + cells increased continuously while both the colony density and the proportion of CD34 + cells monotonically declined with the culture time. During the cultrues of both cell fractions, the proportion of CFU-GM(colony-forming unit granulocyte and macrophage) in CFCs increased continuously, while that of BFU-E(burst-forming unit erythroid) decreased with the culture time. The results showed that the MNC could lead to more colony-forming cells and CD34 + cells during the 3 weeks short-term cultures.