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首页> 外文期刊>Journal of Cellular Physiology >Cultivation of rat marrow-derived mesenchymal stem cells in reduced oxygen tension: effects on in vitro and in vivo osteochondrogenesis.
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Cultivation of rat marrow-derived mesenchymal stem cells in reduced oxygen tension: effects on in vitro and in vivo osteochondrogenesis.

机译：鼠骨髓来源间充质干细胞的培养减少氧张力:细胞的影响体外和体内osteochondrogenesis。

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摘要

Rat mesenchymal stem cells (rMSCs) represent a small portion of the cells in the stromal compartment of bone marrow and have the potential to differentiate into bone, cartilage, fat, and fibrous tissue. These mesenchymal progenitor cells were maintained as primary isolates and as subcultured cells in separate closed modular incubator chambers purged with either 95% air and 5% CO(2) (20% or control oxygen) or 5% oxygen, 5% CO(2), and 90% nitrogen (5% or low oxygen). At first passage, some cells from each oxygen condition were loaded into porous ceramic vehicles and implanted into syngeneic host animals in an in vivo assay for osteochondrogenesis. The remaining cells were continued in vitro in the same oxygen tension as for primary culture or were switched to the alternate condition. The first passage cells were examined for in vitro osteogenesis with assays involving the quantification of alkaline phosphatase activity and calcium and DNA content as well as by von Kossa staining to detect mineralization. Cultures maintained in low oxygen had a greater number of colonies as primary isolates and proliferated more rapidly throughout their time in vitro, as indicated by hemacytometer counts at the end of primary culture and increased DNA values for first passage cells. Moreover, rMSCs cultivated in 5% oxygen produced more bone than cells cultured in 20% oxygen when harvested and loaded into porous ceramic cubes and implanted into syngeneic host animals. Finally, markers for osteogenesis, including alkaline phosphatase activity, calcium content, and von Kossa staining, were elevated in cultures which had been in low oxygen throughout their cultivation time. Expression of these markers was usually increased above basal levels when cells were switched from control to low oxygen at first passage and decreased for cells switched from low to control oxygen. We conclude that rMSCs in culture function optimally in an atmosphere of reduced oxygen that more closely approximates documented in vivo oxygen tension. Copyright 2001 Wiley-Liss, Inc.
机译：大鼠间充质干细胞(rMSCs)表示很小一部分的细胞基质室的骨髓和有潜力分化为骨、软骨、脂肪和纤维组织。细胞作为主要隔离和维护亚文化细胞在独立封闭的模块化孵化室净化空气和与95%有限公司(2)5%(20%或控制氧气)或5%氧气,5%有限公司(2),90%的氮或低氧(5%)。首先,一些细胞从每个氧气条件被加载到多孔陶瓷车辆和植入同源的主机动物的体内试验osteochondrogenesis。继续体外氧张力一样为主要文化或转向备用状态。检查与化验体外骨生成涉及碱性的量化磷酸酶活性和钙和DNA含量和冯Kossa染色检测矿化。有更多的殖民地为主隔离和更迅速地扩散他们的时间在体外,如所示血细胞计数器计数的初选文化和增加DNA值通过细胞。氧气产生比细胞培养在骨头20%的氧气当收获和加载到多孔陶瓷立方体和植入同源的主机动物。包括碱性磷酸酶活性、钙内容,和冯Kossa染色在升高文化一直在低氧他们的培养时间。标记通常是高于基础水平增加当细胞从控制转向低氧气在第一通道和减少细胞从低转向控制氧气。rMSCs文化功能优化减少氧气的气氛接近体内氧张力。版权2001 Wiley-Liss公司。

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