High throughput screening of mesenchymal stromal cell morphological response to inflammatory signals for bioreactor-based manufacturing of extracellular vesicles that modulate microglia.
High throughput screening of mesenchymal stromal cell morphological response to inflammatory signals for bioreactor-based manufacturing of extracellular vesicles that modulate microglia.
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高通量筛选间充质基质细胞对炎症信号的形态反应，用于基于生物反应器制造调节小胶质细胞的细胞外囊泡。
DOI：
10.1101/2023.11.19.567730
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发表时间：
2023
期刊：
影响因子：
--
通讯作者：
Marklein,RossA
中科院分区：
文献类型：
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作者：
Larey,AndrewM;Spoerer,ThomasM;Daga,KanupriyaR;Morfin,MariaG;Hynds,HannahM;Carpenter,Jana;Hines,KellyM;Marklein,RossA
由于其免疫调节功能，间充质基质细胞（MSC）是一种有前途的治疗与神经退行性疾病相关的神经炎症的潜力。该功能由分泌的细胞外囊泡（MSC-EV）介导。尽管已确定安全性，但MSC临床转化由于功能异质性导致的临床结果不一致而不成功。目前减轻功能异质性的方法包括用炎症信号“激发”MSC以增强功能。然而，尚未对预激及其对MSC-EV功能的影响进行全面评价。此外，MSC-EV疗法的临床转化需要显著的生产规模扩大，但很少有研究调查了生物反应器中预充的影响。由于MSC形态学已被证明可以预测其免疫调节功能，因此我们筛选了MSC对一系列引发信号的形态学反应，并评估了MSC-EV在烧瓶和生物反应器中对引发的反应的特性和效力。我们确定了独特的启动条件对应于不同的形态。这些条件证明了一系列MSC-EV制备质量和脂质体，使我们能够发现一种新的MSC-EV制造条件，并深入了解MSC-EV小胶质细胞调节的潜在机制。我们的新型筛选方法和对MSC-EV生物反应器制造的启动应用为大规模制造的改进和MSC-EV功能的增强提供了信息。
Due to their immunomodulatory function, mesenchymal stromal cells (MSCs) are a promising therapeutic with the potential to treat neuroinflammation associated with neurodegenerative diseases. This function is mediated by secreted extracellular vesicles (MSC-EVs). Despite established safety, MSC clinical translation has been unsuccessful due to inconsistent clinical outcomes resulting from functional heterogeneity. Current approaches to mitigate functional heterogeneity include ‘priming’ MSCs with inflammatory signals to enhance function. However, comprehensive evaluation of priming and its effects on MSC-EV function has not been performed. Furthermore, clinical translation of MSC-EV therapies requires significant manufacturing scale-up, yet few studies have investigated the effects of priming in bioreactors. As MSC morphology has been shown to predict their immunomodulatory function, we screened MSC morphological response to an array of priming signals and evaluated MSC-EV identity and potency in response to priming in flasks and bioreactors. We identified unique priming conditions corresponding to distinct morphologies. These conditions demonstrated a range of MSC-EV preparation quality and lipidome, allowing us to discover a novel MSC-EV manufacturing condition, as well as gain insight into potential mechanisms of MSC-EV microglia modulation. Our novel screening approach and application of priming to MSC-EV bioreactor manufacturing informs refinement of larger-scale manufacturing and enhancement of MSC-EV function.