Sun B, Bai C X, Feng K, Li L, Zhao P, Pei X T
Beijing Institute of Transfusion Medicine, Beijing 100850, China.
Sheng Li Xue Bao. 2000 Apr;52(2):143-6.
To elucidate the effects of hypoxia on the proliferation and differentiation of CD34(+) hematopoietic stem/progenitor cells and their response to cytokines, the cells were isolated from umbilical cord blood by using a high-gradient magnetic cell sorting system (MACS). Mononuclear cells (MNC) and CD34(+) cells were incubated in severe hypoxia (1% oxygen) culture system, and the colony forming cells and antigen expression were studied by colony forming assays and FACS analysis. The results showed that incubation in severe hypoxia increased the number of erythroid bursts (BFU-E) (324.8+/-41.4/10(4) cells) generated from CD34(+) cells (191.2+/-34.5/10(4) cells in the control group, P<0.01). Severe hypoxia also enhanced the maintenance and cloning efficiency of BFU-E in a liquid culture system without growth factors, the number of BFU-E (152.4+/-22.6/10(4)cells) was much bigger than that in the control group (74.2+/-9.3/10(4) cells, P<0.01). In cultures incubated in hypoxia, the percentage of CD34(+) cells was significantly higher (2.5+/-1.2-fold, P<0.05) than in those incubated in air. BFU-E cloning efficiency of MNC was not significantly affected by hypoxia. The above results show that hypoxia enhances the maintenance of erythroid progenitor cells generated from CD34(+) hematopoietic stem/progenitor cells, no matter growth factors are present or not. These positive effects of hypoxia did not occur for the other progenitors.
为阐明缺氧对CD34(+)造血干/祖细胞增殖、分化及其对细胞因子反应的影响,采用高梯度磁性细胞分选系统(MACS)从脐带血中分离细胞。将单核细胞(MNC)和CD34(+)细胞置于严重缺氧(1%氧气)培养系统中培养,通过集落形成试验和流式细胞术分析研究集落形成细胞和抗原表达。结果显示,在严重缺氧条件下培养可增加CD34(+)细胞产生的红系爆式集落形成单位(BFU-E)数量(324.8±41.4/10(4)个细胞)(对照组为191.2±34.5/10(4)个细胞,P<0.01)。严重缺氧还提高了无生长因子液体培养系统中BFU-E的维持和克隆效率,BFU-E数量(152.4±22.6/10(4)个细胞)远高于对照组(74.2±9.3/10(4)个细胞,P<0.01)。在缺氧培养的细胞中,CD34(+)细胞的百分比显著高于在空气中培养的细胞(2.5±1.2倍,P<0.05)。缺氧对MNC的BFU-E克隆效率无显著影响。上述结果表明,无论有无生长因子,缺氧均可增强CD34(+)造血干/祖细胞产生的红系祖细胞的维持能力。缺氧对其他祖细胞未产生这些积极作用。