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

Samatha Bhat,Shashank Chandanala,Pachaiyappan Viswanathan,S Jyothi Prasanna,Raviraja N Seetharam

doi:10.1038/s41598-021-83088-1

Samatha Bhat, Shashank Chandanala + Show 3 more

Open Access

https://doi.org/10.1038/s41598-021-83088-1

Copy DOI

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.

Highlights

Bone marrow-derived mesenchymal stromal cells (BM-Mesenchymal stromal cells (MSCs)) are gaining increasing importance in the field of regenerative medicine
Bone marrow-derived mesenchymal stromal cells (BM-MSCs) cultured in low serum/serum-free media (SFM)/XFM at both seeding densities were well adapted to the respective media in the conditions tested and were able to proliferate linearly
The cells grown in StemXVivo SFM at passage 5 took a long time to grow than the previous passage, which could not proliferate beyond 40% confluence

Summary

Introduction

Bone marrow-derived mesenchymal stromal cells (BM-MSCs) are gaining increasing importance in the field of regenerative medicine. 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. 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. We have tested six different commercially available low serum/SFM/XFM for their suitability for the growth and expansion of undifferentiated BM-MSCs (Table 1) in comparison with the serum-containing inhouse medium. We assessed and compared morphological characteristics, population doubling and doubling time, cell size, expression of cell surface markers, colony-forming ability, potency in terms of vascular endothelial growth factor (VEGF) secretion, immunosuppressive and differentiation potential of BM-MSCs cultured in these low serum/SFMs/XFMs as opposed to serum-containing medium

Methods

Results

Conclusion