Hao Yi-Wen, Cheng Da-Ye, Zhou Wen-Ling, Ma Yi-Ran, Wang Yu-Ting
Department of Blood Transfusion, China Medical University First Hospital, Shenyang, 110001, Liaoning Province, China.
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2010 Aug;18(4):997-1001.
Hypoxia in bone marrow is suitable for the perfect preservation of biological functions of bone marrow hematopoietic stem cells (BM HSC). It is deserved to study whether the biological functions of BM HSC are influenced when being exposed to environment of oxygen at various concentration during amplification of BM HSCs in normal oxygen condition in vitro and process of peripheral blood hematopoietic stem cell transplantation (PBSCT). This study was purposed to investigate the effects of various oxygen concentrations on biological functions of human BM HSCs. The BM HSCs were amplified in vitro, the amplification level of CD34(+) HSCs and CD34(+)AC133(+) HSCs were detected by flow cytometry, the apoptosis and cell cycle distribution of CD34(+) HSCs amplified in various oxygen concentrations were assayed by flow cytometry with Annexin V/PI double staining as well as PI and Ki-67 antibody, respectively, the differentiation of amplified CD34(+) HSCs in vitro was determined by direction differentiation assay, the migration ability of amplified CD34(+)AC133(+) HSCs was measured by migration test. The results indicated that the oxygen environment below normal oxygen, especially hypoxia, could amplify more primitive CD34(+)AC133(+) HSCs and CD34(+) HSCs with activity, arrest more HSCs in G₀/G₁ phase, promote the generation of BFU-E, CFU-GM, CFU-GEMM, and better preserve the migration ability of HSCs. While the above functional indicators of BM HSCs were poor when HSCs exposed to normoxia, oxygen-unstable and oxygen-severe changeable environments. It is concluded that the biological functions of BM HSCs in PBSCT are related with oxygen concentration and its stability, the culture of BM HSCs in lower oxygen environment may be more beneficial for PBSCT.
骨髓中的低氧环境适合完美保存骨髓造血干细胞(BM HSC)的生物学功能。在体外正常氧条件下扩增BM HSC以及在外周血造血干细胞移植(PBSCT)过程中,当BM HSC暴露于不同浓度氧气环境时,其生物学功能是否受到影响值得研究。本研究旨在探讨不同氧气浓度对人BM HSC生物学功能的影响。体外扩增BM HSC,通过流式细胞术检测CD34(+) HSC和CD34(+)AC133(+) HSC的扩增水平,分别采用Annexin V/PI双染以及PI和Ki-67抗体通过流式细胞术检测在不同氧气浓度下扩增的CD34(+) HSC的凋亡和细胞周期分布,通过定向分化试验测定体外扩增的CD34(+) HSC的分化情况,通过迁移试验检测扩增的CD34(+)AC133(+) HSC的迁移能力。结果表明,低于正常氧的氧气环境,尤其是低氧环境,能够扩增出更多具有活性的原始CD34(+)AC133(+) HSC和CD34(+) HSC,使更多的HSC停滞在G₀/G₁期,促进爆式红系集落形成单位(BFU-E)、粒-巨噬细胞集落形成单位(CFU-GM)、粒-红-巨噬-巨核系集落形成单位(CFU-GEMM)的生成,并更好地保留HSC的迁移能力。而当HSC暴露于常氧、氧不稳定和氧剧烈变化环境时,BM HSC的上述功能指标较差。结论:PBSCT中BM HSC的生物学功能与氧气浓度及其稳定性有关,在较低氧气环境中培养BM HSC可能对PBSCT更有益。