Jeske Richard, Lewis Shaquille, Tsai Ang-Chen, Sanders Kevin, Liu Chang, Yuan Xuegang, Li Yan
Department of Chemical and Biomedical Engineering, FAMU-FSU College of Engineering, Florida State University, Tallahassee, Florida, United States.
The National High Magnetic Field Laboratory, Florida State University, Tallahassee, Florida.
Biochem Eng J. 2021 Apr;168. doi: 10.1016/j.bej.2021.107947. Epub 2021 Jan 27.
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 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.
人间充质干细胞(hMSCs)因其分泌营养因子、多能分化潜能和自我更新能力而在细胞治疗中广为人知。因此,在过去十年中,临床试验的数量一直在稳步增加,这凸显了对能够大量生产细胞以满足不断增长需求的体外系统的需求。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,这可能导致细胞内稳态的改变。在本研究中,hMSCs在125 mL转瓶生物反应器系统中的微载体上进行扩增。我们的结果表明,在七天内细胞扩增了三倍。此外,我们的结果表明,与静态平面培养相比,在基于微载体的悬浮生物反应器中培养hMSCs会导致细胞尺寸更小,活性氧(ROS)和ROS调节因子Sirtuin-3水平更高,这对烟酰胺腺嘌呤二核苷酸代谢途径和代谢稳态有影响。此外,生物反应器中的hMSCs在受到γ干扰素刺激后,前列腺素E分泌增加,吲哚胺-吡咯2,3-双加氧酶分泌减少。本研究结果有助于理解在hMSCs用于生物制造和临床试验的可扩展生产过程中,生物反应器微环境对hMSCs潜在生理变化的影响。