Physiologically Low Oxygen Enhances Biomolecule Production and Stemness of Mesenchymal Stem Cell Spheroids.
Physiologically Low Oxygen Enhances Biomolecule Production and Stemness of Mesenchymal Stem Cell Spheroids.
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DOI：
10.1089/ten.tec.2015.0465
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发表时间：
2016-03
期刊：
影响因子：
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通讯作者：
Emily R. Shearier;Qi Xing;Zichen Qian;F. Zhao
中科院分区：
文献类型：
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作者：
Emily R. Shearier;Qi Xing;Zichen Qian;F. Zhao
多细胞人间充质干细胞（hMSC）球体已被证明在多种应用中具有价值，包括软骨再生、伤口愈合和新血管生成。生理相关低氧培养可通过阻止细胞分化而显著提高hMSC的体外扩增。我们推测低氧培养的hMSC球体可以更好地保持hMSC的再生特性。本研究采用悬滴法制备hMSC球体，并在2%O2和20%O2下培养96 h。球体直径和活力进行了检查，以及细胞外基质（ECM）成分和生长因子水平之间的两个氧分压在不同的时间点。在球状体培养条件下测量干度，并与二维细胞培养物进行比较。使用不同的针规研究球体活力和结构完整性，以确保在体内实施时不会发生损坏。还证明了组织替代物内的球体附着和整合。结果表明，在低氧条件下培养的三维hMSC球体可以增强ECM蛋白和生长因子的产生，同时保持球体的干性和被注射、附着和潜在地整合在组织内的能力。
Multicellular human mesenchymal stem cell (hMSC) spheroids have been demonstrated to be valuable in a variety of applications, including cartilage regeneration, wound healing, and neoangiogenesis. Physiological relevant low oxygen culture can significantly improve in vitro hMSC expansion by preventing cell differentiation. We hypothesize that hypoxia-cultured hMSC spheroids can better maintain the regenerative properties of hMSCs. In this study, hMSC spheroids were fabricated using hanging drop method and cultured under 2% O2 and 20% O2 for up to 96 h. Spheroid diameter and viability were examined, as well as extracellular matrix (ECM) components and growth factor levels between the two oxygen tensions at different time points. Stemness was measured among the spheroid culture conditions and compared to two-dimensional cell cultures. Spheroid viability and structural integrity were studied using different needle gauges to ensure no damage would occur when implemented in vivo. Spheroid attachment and integration within a tissue substitute were also demonstrated. The results showed that a three-dimensional hMSC spheroid cultured at low oxygen conditions can enhance the production of ECM proteins and growth factors, while maintaining the spheroids' stemness and ability to be injected, attached, and potentially be integrated within a tissue.