Expansion and characterization of bone marrow derived human mesenchymal stromal cells in serum-free conditions

Abstract

Bone marrow-derived mesenchymal stromal cells (BM-MSCs) are gaining increasing importance in the field of regenerative medicine. Although therapeutic value of MSCs is now being established through many clinical trials, issues have been raised regarding their expansion as per regulatory guidelines. Fetal bovine serum usage in cell therapy poses difficulties due to its less-defined, highly variable composition and safety issues. Hence, there is a need for transition from serum-based to serum-free media (SFM). Since SFM are cell type-specific, a precise analysis of the properties of MSCs cultured in SFM is required to determine the most suitable one. Six different commercially available low serum/SFM with two different seeding densities were evaluated to explore their ability to support the growth and expansion of BM-MSCs and assess the characteristics of BM-MSCs cultured in these media. Except for one of the SFM, all other media tested supported the growth of BM-MSCs at a low seeding density. No significant differences were observed in the expression of MSC specific markers among the various media tested. In contrary, the population doubling time, cell yield, potency, colony-forming ability, differentiation potential, and immunosuppressive properties of MSCs varied with one another. We show that SFM tested supports the growth and expansion of BM-MSCs even at low seeding density and may serve as possible replacement for animal-derived serum.

Antibacterial and Osteogenic Properties of Ag Nanoparticles and Ag/TiO2 Nanostructures Prepared by Atomic Layer Deposition

“…FetMSCs on the surface of all the samples are elongated, bipolar, spindle-shaped, and interconnected by filopodia ( Figure 8 e–h). This morphology of the cells is typical for the cells of this line [ 28 , 29 ]. The greatest number of cells was on the surface of the polished titanium.…”

Heterogeneity of In Vitro Expanded Mesenchymal Stromal Cells and Strategies to Improve Their Therapeutic Actions

“…However, FBS may contain animal derivatives enhancing the risk of zoonotic transmission and immunological reactions during cell therapy. In this context, human platelet preparations, such as platelet lysate or platelet-rich plasma, have emerged as an alternative to the use of non-human serum, favoring the biological safety of MSC therapies [ 88 , 89 , 90 ]. Importantly, the clonogenic efficiency and proliferative capacity of MSCs was found to be greater when using 5% human platelet lysate as compared to 10% fetal serum, probably due to the high concentration of natural growth factors contained in platelets [ 90 ].…”

When Origin Matters: Properties of Mesenchymal Stromal Cells From Different Sources for Clinical Translation in Kidney Disease

“…To date, 10% FCS continues to be the most common media supplement employed in clinical trials for kidney disease, although some consideration has been given to the use of xenogeneic-free media such as hPL (5 or 10%), human serum albumin (HSA) and a CDM (Table 2). Currently, some commercial and non-commercial CDM formulations have been investigated (359)(360)(361)(362)(363), however, there is still limited availability of some of these media for large-scale manufacturing at GMP quality level.…”