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Open Access Highly Accessed Research

A novel serum-free medium for the expansion of human mesenchymal stem cells

Lucas G Chase1*, Uma Lakshmipathy2, Luis A Solchaga3, Mahendra S Rao4 and Mohan C Vemuri4

Author Affiliations

1 Primary and Stem Cell Systems, Life Technologies, 501 Charmany Drive, Madison, WI 53719, USA

2 Primary and Stem Cell Systems, 5781 Van Allen Way, Carlsbad, CA 92008, USA

3 Department of General Medical Sciences, Division of Hematology and Oncology, Case Western Reserve University, 2103 Cornell Road, Cleveland, OH 44106, USA

4 Primary and Stem Cell Systems, Life Technologies, 7335 Executive Way, Frederick, MD 21704, USA

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Stem Cell Research & Therapy 2010, 1:8  doi:10.1186/scrt8

Published: 2 April 2010

Abstract

Introduction

Human multipotent mesenchymal stem cell (MSC) therapies are being tested clinically for a variety of disorders, including Crohn's disease, multiple sclerosis, graft-versus-host disease, type 1 diabetes, bone fractures, and cartilage defects. However, despite the remarkable clinical advancements in this field, most applications still use traditional culture media containing fetal bovine serum. The ill-defined and highly variable nature of traditional culture media remains a challenge, hampering both the basic and clinical human MSC research fields. To date, no reliable serum-free medium for human MSCs has been available.

Methods

In this study, we developed and tested a serum-free growth medium on human bone marrow-derived MSCs through the investigation of multiple parameters including primary cell isolation, multipassage expansion, mesoderm differentiation, cellular phenotype, and gene-expression analysis.

Results

Similar to that achieved with traditional culture medium, human MSCs expanded in serum-free medium supplemented with recombinant human platelet-derived growth factor-BB (PDGF-BB), basic fibroblast growth factor (bFGF), and transforming growth factor (TGF)-β1 showed extensive propagation with retained phenotypic, differentiation, and colony-forming unit potential. To monitor global gene expression, the transcriptomes of bone marrow-derived MSCs expanded under serum-free and serum-containing conditions were compared, revealing similar expression profiles. In addition, the described serum-free culture medium supported the isolation of human MSCs from primary human marrow aspirate with continual propagation.

Conclusions

Although the described serum-free MSC culture medium is not free of xenogeneic components, this medium provides a substitute for serum-containing medium for research applications, setting the stage for future clinical applications.