Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory University, Atlanta, Georgia.
Emory University School of Medicine, Winship Cancer Institute, Atlanta, Georgia.
Biotechnol Bioeng. 2020 Jun;117(6):1761-1778. doi: 10.1002/bit.27317. Epub 2020 Mar 9.
Mesenchymal stromal cells (MSCs) have failed to consistently demonstrate their therapeutic efficacy in clinical trials, due in part to variability in culture conditions used for their production. Of various culture conditions used for MSC production, aggregate culture has been shown to improve secretory capacity (a putative mechanism of action in vivo) compared with standard monolayer culture. The purpose of this study was to perform multiomics characterization of MSCs cultured in monolayer and as aggregates to identify aspects of cell physiology that differ between these culture conditions to begin to understand cellular-level changes that might be related to secretory capacity. Targeted secretome characterization was performed on multiple batches of MSC-conditioned media, while nontargeted proteome and metabolome characterization was performed and integrated to identify cellular processes differentially regulated between culture conditions. Secretome characterization revealed a reduction in MSC batch variability when cultured as aggregates. Proteome and metabolome characterization showed upregulation of multiple protein and lipid metabolic pathways, downregulation of several cytoskeletal processes, and differential regulation of extracellular matrix synthesis. Integration of proteome and metabolome characterization revealed individual lipid metabolites and vesicle-trafficking proteins as key features for discriminating between culture conditions. Overall, this study identifies several aspects of MSC physiology that are altered by aggregate culture. Further exploration of these processes and pathways is needed to determine their potential role in regulating cell secretory capacity.
间充质基质细胞（MSCs）在临床试验中未能始终如一地证明其治疗效果，部分原因是用于其生产的培养条件存在差异。在用于 MSC 生产的各种培养条件中，与标准单层培养相比，聚集体培养已被证明可提高分泌能力（体内的一种假定作用机制）。本研究的目的是对单层和聚集体培养的 MSC 进行多组学特征分析，以确定这些培养条件之间在细胞生理学方面的差异，从而开始了解可能与分泌能力相关的细胞水平变化。对多个批次的 MSC 条件培养基进行了靶向分泌组学特征分析，同时对非靶向蛋白质组学和代谢组学特征进行了分析和整合，以鉴定培养条件下差异调节的细胞过程。聚集体培养时，MSC 批次变异性降低。蛋白质组学和代谢组学特征表明，多个蛋白质和脂质代谢途径上调，几个细胞骨架过程下调，细胞外基质合成的差异调节。蛋白质组学和代谢组学特征的整合揭示了个体脂质代谢物和囊泡运输蛋白作为区分培养条件的关键特征。总的来说，这项研究确定了聚集体培养改变的 MSC 生理学的几个方面。需要进一步探索这些过程和途径，以确定它们在调节细胞分泌能力方面的潜在作用。
