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 guid...

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Main Authors: Samatha Bhat, Pachaiyappan Viswanathan, Shashank Chandanala, S. Jyothi Prasanna, Raviraja N. Seetharam
Format: Article
Language:English
Published: Nature Publishing Group 2021-02-01
Series:Scientific Reports
Online Access:https://doi.org/10.1038/s41598-021-83088-1
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spelling doaj-507e5f051ff34c1ba30abd9f3c42903b2021-02-14T12:36:07ZengNature Publishing GroupScientific Reports2045-23222021-02-0111111810.1038/s41598-021-83088-1Expansion and characterization of bone marrow derived human mesenchymal stromal cells in serum-free conditionsSamatha Bhat0Pachaiyappan Viswanathan1Shashank Chandanala2S. Jyothi Prasanna3Raviraja N. Seetharam4Stempeutics Research Pvt Ltd, Shirdi Sai Baba Cancer HospitalStempeutics Research Pvt Ltd, Shirdi Sai Baba Cancer HospitalManipal Institute of Regenerative Medicine, Manipal Academy of Higher EducationManipal Institute of Regenerative Medicine, Manipal Academy of Higher EducationStempeutics Research Pvt Ltd, Shirdi Sai Baba Cancer HospitalAbstract 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.https://doi.org/10.1038/s41598-021-83088-1
collection DOAJ
language English
format Article
sources DOAJ
author Samatha Bhat
Pachaiyappan Viswanathan
Shashank Chandanala
S. Jyothi Prasanna
Raviraja N. Seetharam
spellingShingle Samatha Bhat
Pachaiyappan Viswanathan
Shashank Chandanala
S. Jyothi Prasanna
Raviraja N. Seetharam
Expansion and characterization of bone marrow derived human mesenchymal stromal cells in serum-free conditions
Scientific Reports
author_facet Samatha Bhat
Pachaiyappan Viswanathan
Shashank Chandanala
S. Jyothi Prasanna
Raviraja N. Seetharam
author_sort Samatha Bhat
title Expansion and characterization of bone marrow derived human mesenchymal stromal cells in serum-free conditions
title_short Expansion and characterization of bone marrow derived human mesenchymal stromal cells in serum-free conditions
title_full Expansion and characterization of bone marrow derived human mesenchymal stromal cells in serum-free conditions
title_fullStr Expansion and characterization of bone marrow derived human mesenchymal stromal cells in serum-free conditions
title_full_unstemmed Expansion and characterization of bone marrow derived human mesenchymal stromal cells in serum-free conditions
title_sort expansion and characterization of bone marrow derived human mesenchymal stromal cells in serum-free conditions
publisher Nature Publishing Group
series Scientific Reports
issn 2045-2322
publishDate 2021-02-01
description 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.
url https://doi.org/10.1038/s41598-021-83088-1
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