Agitation in a microcarrier-based spinner flask bioreactor modulates homeostasis of human mesenchymal stem cells
Agitation in a microcarrier-based spinner flask bioreactor modulates homeostasis of human mesenchymal stem cells
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DOI：
10.1016/j.bej.2021.107947
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发表时间：
2021-02-02
影响因子：
3.9
通讯作者：
Li, Yan
中科院分区：
文献类型：
--
作者：
Jeske, Richard;Lewis, Shaquille;Li, Yan
人间充质干细胞（hMSC）由于其分泌营养因子、多能分化潜能和自我更新能力而在细胞治疗中众所周知。因此，在过去十年中，临床试验的数量一直在稳步增加，突出了对能够产生大量细胞的体外系统的需求，以满足不断增长的需求。然而，hMSCs对微环境条件高度敏感，包括动态生物反应器系统引起的剪切应力，并可导致细胞稳态的改变。在这项研究中，hMSC在125 mL转瓶生物反应器系统内的微载体上扩增。我们的研究结果表明，在七天内扩增了三倍。此外，我们的研究结果表明，在基于微载体的悬浮生物反应器中培养hMSCs（与静态平面培养相比）导致更小的细胞尺寸和更高水平的活性氧化物质（ROS）和ROS调节剂Sirtuin-3，这对烟酰胺腺嘌呤二核苷酸代谢途径和代谢稳态具有影响。此外，生物反应器中的hMSC显示出在干扰素γ刺激后增加的前列腺素E-2分泌以及减少的吲哚胺-吡咯2，3-双加氧酶分泌。本研究的结果提供了在用于生物制造和临床试验的hMSC的可规模生产期间受生物反应器微环境影响的潜在hMSC生理学改变的理解。
Human mesenchymal stem cells (hMSCs) are well known in cell therapy due to their secretion of trophic factors, multipotent differentiation potential, and ability for self-renewal. As a result, the number of clinical trials has been steadily increasing over the last decade highlighting the need for in vitro systems capable of producing large quantities of cells to meet growing demands. However, hMSCs are highly sensitive to microenvironment conditions, including shear stress caused by dynamic bioreactor systems, and can lead to alteration of cellular homeostasis. In this study, hMSCs were expanded on microcarriers within a 125 mL spinner flask bioreactor system. Our results demonstrate a three-fold expansion over seven days. Furthermore, our results show that culturing hMSCs in the microcarrier-based suspension bioreactor (compared to static planar culture) results in smaller cell size and higher levels of reactive oxidative species (ROS) and ROS regulator Sirtuin-3, which have implications on the nicotinamide adenine dinucleotide metabolic pathway and metabolic homeostasis. In addition, hMSCs in the bioreactor showed the increased Prostaglandin E-2 secretion as well as reduced the Indoleamine-pyrrole 2,3-dioxygenase secretion upon stimulus with interferon gamma. The results of this study provide understanding of potential hMSC physiology alterations impacted by bioreactor microenvironment during scalable production of hMSCs for biomanufacturing and clinical trials.