Yu Yang, Lin Lixiang, Zhou Yifei, Lu Xiaolang, Shao Xiwen, Lin Chuanlu, Yu Kehe, Zhang Xiaolei, Hong Jianjun, Chen Ying
Department of Orthopaedics Surgery, The 2nd Affiliated Hospital and Yuying Children's Hospital of Wenzhou Medical University, Wenzhou, Zhejiang, China (mainland).
Department of Orthopaedic Surgery, The 2nd Affiliated Hospital and Yuying Children's Hospital of Wenzhou Medical University, Wenzhou, Zhejiang, China (mainland).
Med Sci Monit. 2017 Mar 17;23:1334-1339. doi: 10.12659/msm.903892.
BACKGROUND Hypoxic conditions play roles in functioning of human tendon-derived stem cells (hTSCs). The goal of this study was to investigate the effect of various hypoxic conditions in self-renewal capacity and differentiation of TSCs. MATERIAL AND METHODS hTSCs was obtain from supraspinatus tendon donors. Colony formation and cell proliferation assay were used to assess the self-renewal of hTSCs. qRT-PCT and Western blot analysis were used to examine stemness and multi-differentiation potential of hTSCs. RESULTS We found that culturing at 5% O2 is more beneficial for the self-renewal of hTSCs than the other 3 culture conditions, with larger colony size and numbers. The proliferation of hTSCs in 5%, 10%, and 20% O2 cultures increased after seeding. The number of cells in the 5% O2 condition was higher than that in other culture; however, self-renewal capacity of hTSCs in 0.5% O2 was inhibited. The expression levels of stem cell markers, including NS, Nanog, Oct-4, and SSEA-4, were highest in 0.5% O2 culture. Furthermore, hTSCs cultured in 20% O2 exhibited significantly higher expression of the 3 markers (PPAR-γ, Sox-9, and Runx-2). CONCLUSIONS Hypoxic condition of culture encouraged self-renewal capacity of hTSCs, but inhibited their multi-differentiation potential, compared to normoxic condition of culture. Moreover, excessively low oxygen concentration impaired the capacity of hTSCs.
背景 低氧条件在人肌腱衍生干细胞（hTSCs）的功能中发挥作用。本研究的目的是探讨不同低氧条件对TSCs自我更新能力和分化的影响。 材料与方法 从冈上肌腱供体获取hTSCs。采用集落形成和细胞增殖试验评估hTSCs的自我更新能力。采用qRT - PCT和蛋白质印迹分析检测hTSCs的干性和多分化潜能。 结果 我们发现，与其他3种培养条件相比，在5%氧气浓度下培养对hTSCs的自我更新更有益，集落更大且数量更多。接种后，hTSCs在5%、10%和20%氧气浓度培养中的增殖增加。5%氧气浓度条件下的细胞数量高于其他培养条件；然而，0.5%氧气浓度下hTSCs的自我更新能力受到抑制。包括NS、Nanog、Oct - 4和SSEA - 4在内的干细胞标志物的表达水平在0.5%氧气浓度培养中最高。此外，在20%氧气浓度下培养的hTSCs表现出3种标志物（PPAR - γ、Sox - 9和Runx - 2）的表达显著更高。 结论 与正常氧浓度培养相比，培养的低氧条件促进了hTSCs的自我更新能力，但抑制了它们的多分化潜能。此外，过低的氧浓度损害了hTSCs的能力。
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