Phenotypic and functional comparison of optimum culture conditions for upscaling of bone marrow-derived mesenchymal stem cells

Abstract

Human adult bone marrow-derived mesenchymal stem cells (MSCs) are a promising tool in the newly emerging avenue of regenerative medicine. MSCs have already been translated from basic research to clinical transplantation research. However, there is still a lack of consensus on the ideal method of culturing MSCs. Here we have compared different culture conditions of human MSCs with an attempt to preserve their characteristics and multi-lineage differentiation potential. We compare the different basal culture media DMEM-F12, DMEM-high glucose (DMEM-HG), DMEM-low glucose (DMEM-LG), knock-out DMEM (DMEM-KO) and Mesencult on the proliferation rate, surface markers and differentiation potentials of MSCs. At every fifth passage until the 25th passage, the differentiation potential and the presence of a panel of surface markers was observed, using flow cytometry. We also compared the characteristics of human MSCs when cultured in reduced concentrations of fetal bovine serum (FBS), knockout serum replacement (KO-SR) and human plasma. Data indicate that the presence of serum is essential to sustain and propagate MSCs cultures. The choice of basal medium is equally important so as to preserve their characteristics and multipotent properties even after prolonged culture in vitro. With MSCs emerging as a popular tool for regenerative therapies in incurable diseases, it is essential to be able to obtain a large number of MSCs that continue to preserve their characteristics following passaging. The data reveal the optimum basal medium for prolonged culture of MSCs while retaining their ability to differentiate and hence this may be used for up-scaling to provide sufficient numbers for transplantation.

How has dental pulp stem cells isolation been conducted? A scoping review

“…33 Our data showed that nearly 95% of the articles described the culture medium used for DPSCs (Table 1), indicating a very low risk of bias and therefore highlighting the relevance of this factor. Nineteen different types of culture medium were cited.…”

Culture of equine bone marrow mononuclear fraction and adipose tissue-derived stromal vascular fraction cells in different media

“…al 2012. However, to avoid the potential risk of exposing human patients to cells cultivated in the presence of FBS, some investigators have searched for a substitute supplement (Bieback et al 2009) because, according to the results of Pal et al (2009), the presence of serum is essential for maintaining and expanding mesenchymal stem cell cultures. In the present work, despite samples coming from adipose tissue, they showed greater cellularity in DMEM supplemented with FBS in the first two weeks, but afterwards, there is no longer any difference in relation to grown in AIM-V medium without FBS, suggesting that this can be an alternative for the culture of cells destined for transplantation.…”

Bridging defects in chronic spinal cord injury using peripheral nerve grafts combined with a chitosan-laminin scaffold and enhancing regeneration through them by co-transplantation with bone-marrow-derived mesenchymal stem cells: Case series of 14 patients

“…85 To account for these contradictory reports, it is hypothesized that improved transmission in intact fibers subserving the zone of partial preservation leads to early improvements. 86 Another possible explanation is the source, 87,88 culture conditions, and passage number [89][90][91][92] of the mesenchymal stem cells used, as all are known to influence mesenchymal stem cell potential. A third possible explanation is differential efficacy of mesenchymal stem cells from different donors.…”