Abstract
Background & Aim For the successful development of production effective stem cell therapy, mass production of consistent quality cells is required. Excellent culture medium is important in mass production of quality cells. Classically, fetal bovine serum (FBS) has been used as culture supplement for stem cells. However, due to the undefined and heterologous composition of FBS, researchers have transitioned to more chemically defined serum-free media. In this study, we aimed to investigate and compare the stem cell characteristics cultured in both media as well as media performance tests. Methods, Results & Conclusion Methods Media performance was compared with cell growth rate, accumulated cell number, population homogeneity and cell viability. Comparative analysis of adipose derived mesenchymal stem cells (ADSCs) was performed based on stem cell surface marker expression, differentiation potency, genetic stability, senescence analysis, mRNA expression change, and protein expression change. Results Culture using SFCDM provided more stable population doubling time (PDT), more homogenous cell population, and more cells in a shorter time compared to FBS containing media (FBSCM). The cultured ADSCs were shown to be positive for CD73, CD90, and CD105, but negative for CD14, CD34, CD45, and HLA-DR in both FBSCM and SFCDM. Adipogenic differentiation capability was similar in both media groups, but chondrogenic and osteogenc differentiation capabilities of ADSCs cultured in SFCDM were higher than those cultured in FBSCM. ADSCs positive for β-galactosidase were about 3 times higher in FBSCM . These results indicate that culture in SFCDM has an advantage in reducing cellular senescence of ADSCs compared to FBS media. ADSCs cultured in FBSCM showed a significant increase in the formation frequency of nucleoplasmic bridges comparable to those cultured in SFCDM, suggesting that culture using SFCDM is more genetically stable. The mRNA expression levels of ADSCs cultured in FBSCM were significantly increased for KEGG classification of aging, apoptotic process, cell death, immune response and inflammatory response compared to SFCDM. Conclusion SFCDM can provide higher-speed cell production rate than the classical FBSCM. SFCDM enables the maintenance of higher population homogeneity, genetic stability, and excellent differentiation potency. Taken together, these results suggest that culture using SFCDM provides various advantages through which it is possible to obtain safer, stable, and larger amounts of ADSCs.
Published Version
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