Manipal University, MAHE, Manipal Institute of Regenerative Medicine, GKVK Post, Bellary Road, Allalasandra, Near Royal Orchid, Yelahanka, Bangalore, 560 065, India.
Cell Biol Int. 2013 Nov;37(11):1197-204. doi: 10.1002/cbin.10154. Epub 2013 Aug 13.
Low level of oxygen at the site of injury is likely to affect the viability and proliferation of the transplanted mesenchymal stromal cells (MSCs). Hence there is a need to understand the effect of the physical environment on transplanted stromal cells. Therefore, we have studied the effect of the duration of hypoxic exposure alone or in combination with normoxia on placenta derived mesenchymal stem cell (PDMSCs). PDMSCs and bone marrow MSCs (BMMSCs) were analysed under four different culture conditions, exposure to direct normoxia (N), direct hypoxia (H) and intermittent normoxia followed by hypoxia (NH) and intermittent hypoxia followed by normoxia (HN). The effect on morphology, proliferation, metabolic activity by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide) and viability by 7AAD (7-amino-actinomycin D) were assayed, along with markers for MSCs and HLADR. No change in morphology, marker expression or HLADR was detected in N, H, NH or HN. An increase in proliferation rate, decrease in population doubling-time (PDT) and a relative increase in metabolic activity was strongly noted in the order: NH, N/HN and H. No significant difference was observed in the viability between N, H, NH or HN. A similar pattern was also observed in BMMSCS, indicating comparable suitability of PDMSCs in therapeutic applications. Thus we conclude that intermittent exposure to normoxia prior to hypoxic exposure is a better option than direct exposure to hypoxia. This may have clinical relevance in that they probably mirror the in vivo scenario of systemic delivery (NH) of cells as opposed to local delivery (H), thereby suggesting that systemic delivery is better than local delivery.
损伤部位的低氧水平可能会影响移植间充质基质细胞（MSCs）的活力和增殖。因此，有必要了解物理环境对移植基质细胞的影响。因此，我们研究了单纯缺氧暴露或联合常氧暴露对胎盘来源间充质干细胞（PDMSCs）的影响。PDMSCs 和骨髓间充质干细胞（BMMSCs）在四种不同的培养条件下进行分析，分别为直接常氧（N）、直接缺氧（H）、缺氧后间歇性常氧（NH）和常氧后间歇性缺氧（HN）。通过 MTT（3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐）检测形态、增殖、代谢活性和 7AAD（7-氨基放线菌素 D）检测活力，并检测 MSCs 和 HLA-DR 的标志物。在 N、H、NH 或 HN 中未检测到形态、标志物表达或 HLA-DR 的变化。增殖率增加、群体倍增时间（PDT）缩短和代谢活性相对增加在 NH、N/HN 和 H 中表现最为明显。在 N、H、NH 或 HN 之间未观察到活力有显著差异。BMMSCS 中也观察到类似的模式，表明 PDMSCs 在治疗应用中具有相似的适用性。因此，我们得出结论，与直接缺氧暴露相比，在缺氧暴露前间歇性暴露于常氧是更好的选择。这在临床上可能具有相关性，因为它们可能反映了全身输送（NH）细胞而不是局部输送（H）的体内情况，从而表明全身输送优于局部输送。
