Korntner Stefanie H, Di Nubila Alessia, Gaspar Diana, Zeugolis Dimitrios I
Regenerative, Modular and Developmental Engineering Laboratory (REMODEL) and Science Foundation Ireland (SFI) Centre for Research in Medical Devices (CÚRAM), University of Galway, Galway, Ireland.
Regenerative, Modular and Developmental Engineering Laboratory (REMODEL), Charles Institute of Dermatology, Conway Institute of Biomolecular and Biomedical Research and School of Mechanical and Materials Engineering, University College Dublin, Dublin, Ireland.
Front Bioeng Biotechnol. 2023 Mar 6;11:1136827. doi: 10.3389/fbioe.2023.1136827. eCollection 2023.
Cell culture media containing undefined animal-derived components and prolonged culture periods in the absence of native extracellular matrix result in phenotypic drift of human bone marrow stromal cells (hBMSCs). Herein, we assessed whether animal component-free (ACF) or xeno-free (XF) media formulations maintain hBMSC phenotypic characteristics more effectively than foetal bovine serum (FBS)-based media. In addition, we assessed whether tissue-specific extracellular matrix, induced macromolecular crowding (MMC) during expansion and/or differentiation, can more tightly control hBMSC fate. Cells expanded in animal component-free media showed overall the highest phenotype maintenance, as judged by cluster of differentiation expression analysis. Contrary to FBS media, ACF and XF media increased cellularity over time in culture, as measured by total DNA concentration. While MMC with Ficoll™ increased collagen deposition of cells in FBS media, FBS media induced significantly lower collagen synthesis and/or deposition than the ACF and XF media. Cells expanded in FBS media showed higher adipogenic differentiation than ACF and XF media, which was augmented by MMC with Ficoll™ during expansion. Similarly, Ficoll™ crowding also increased chondrogenic differentiation. Of note, donor-to-donor variability was observed for collagen type I deposition and trilineage differentiation capacity of hBMSCs. Collectively, our data indicate that appropriate screening of donors, media and supplements, in this case MMC agent, should be conducted for the development of clinically relevant hBMSC medicines.
含有成分不明确的动物源性成分的细胞培养基，以及在缺乏天然细胞外基质的情况下延长培养时间，会导致人骨髓间充质干细胞（hBMSCs）的表型漂移。在此，我们评估了无动物成分（ACF）或无异种成分（XF）的培养基配方是否比基于胎牛血清（FBS）的培养基更有效地维持hBMSC的表型特征。此外，我们评估了组织特异性细胞外基质在扩增和/或分化过程中诱导的大分子拥挤（MMC）是否能更严格地控制hBMSC的命运。通过分化簇表达分析判断，在无动物成分培养基中扩增的细胞总体上表现出最高的表型维持率。与FBS培养基相反，通过总DNA浓度测量，ACF和XF培养基在培养过程中细胞数量随时间增加。虽然用聚蔗糖™进行的MMC增加了FBS培养基中细胞的胶原蛋白沉积，但FBS培养基诱导的胶原蛋白合成和/或沉积明显低于ACF和XF培养基。在FBS培养基中扩增的细胞比ACF和XF培养基表现出更高的成脂分化，在扩增过程中用聚蔗糖™进行的MMC增强了这种分化。同样，聚蔗糖™拥挤也增加了软骨形成分化。值得注意的是，观察到hBMSCs的I型胶原蛋白沉积和三系分化能力存在供体间差异。总体而言，我们的数据表明，在开发临床相关的hBMSC药物时，应进行供体、培养基和补充剂（在这种情况下为MMC剂）的适当筛选。
