1. A defined medium and substrate for expansion of human mesenchymal stromal cell progenitors that enriches for osteo- and chondrogenic precursors.
- Author
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Hudson JE, Mills RJ, Frith JE, Brooke G, Jaramillo-Ferrada P, Wolvetang EJ, and Cooper-White JJ
- Subjects
- Adolescent, Adult, Animals, Antigens, Surface metabolism, Cattle, Cell Differentiation drug effects, Cell Lineage drug effects, Cell Proliferation drug effects, Cell Shape drug effects, Chondrogenesis genetics, Colony-Forming Units Assay, Extracellular Matrix drug effects, Extracellular Matrix metabolism, Flow Cytometry, Gene Expression Regulation drug effects, Humans, Mesenchymal Stem Cells metabolism, Osteogenesis genetics, RNA, Messenger genetics, RNA, Messenger metabolism, Stromal Cells cytology, Stromal Cells drug effects, Stromal Cells metabolism, Tissue Donors, Transcription Factors metabolism, Young Adult, Chondrogenesis drug effects, Culture Media pharmacology, Mesenchymal Stem Cells cytology, Mesenchymal Stem Cells drug effects, Osteogenesis drug effects
- Abstract
Human mesenchymal stromal cells (hMSCs) have generated significant interest due to their potential use in clinical applications. hMSCs are present at low frequency in vivo, but after isolation can be expanded considerably, generating clinically useful numbers of cells. In this study, we demonstrate the use of a defined embryonic stem cell expansion medium, mTeSR (Stem Cell Technologies), for the expansion of bone-marrow-derived hMSCs. The hMSCs grow at comparable rates, demonstrate tri-lineage differentiation potential, and show similar surface marker profiles (CD29(+), CD44(+), CD49a(+), CD73(+), CD90(+), CD105(+), CD146(+), CD166(+), CD34(-), and CD45(-)) in both the fetal bovine serum (FBS)-supplemented medium and mTeSR. However, expression of early differentiation transcription factors runt-related transcription factor 2, sex-determining region Y box 9, and peroxisome proliferator-activated receptor gamma changed significantly. Both runt-related transcription factor 2 and sex-determining region Y box 9 were upregulated, whereas peroxisome proliferator-activated receptor gamma was downregulated in mTeSR compared with FBS. Although osteogenic and chondrogenic differentiation was comparable in cells grown in mTeSR compared to FBS, adipogenic differentiation was significantly decreased in mTeSR-expanded cells, both in terms of gene expression and absolute numbers of adipocytes. The removal of the FBS from the medium and the provision of a defined medium with disclosed composition make mTeSR a superior study platform for hMSC biology in a controlled environment. Further, this provides a key step toward generating a clinical-grade medium for expansion of hMSCs for clinical applications that rely on osteo- and chondroinduction of MSCs, such as bone repair and cartilage generation.
- Published
- 2011
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