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Reduced Serum and Hypoxic Culture Conditions Enhance the Osteogenic Potential of Human Mesenchymal Stem Cells

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Abstract

Current protocols for inducing osteogenic differentiation in mesenchymal stem/stromal cells (MSCs) in culture for tissue engineering applications depend on the use of biochemical supplements. However, standard in vitro culture conditions expose cells to ambient oxygen concentrations and high levels of serum (21 % O2, 10 % FBS) that do not accurately recapitulate the physiological milieu. While we and others have examined MSC behavior under hypoxia, the synergistic effect of low serum levels, such as those present in ischemic injury sites, on osteogenic differentiation has not been clearly examined. We hypothesized that a concomitant reduction of serum and O2 would enhance in vitro osteogenic differentiation of MSCs by more accurately mimicking the fracture microenvironment. We show that serum deprivation, in conjunction with hypoxia, potentiates osteogenic differentiation in MSCs. These data demonstrate the role of serum levels in regulating osteogenesis and its importance in optimizing MSC differentiation strategies.

Highlights

  • Serum levels, in addition to hypoxia, have a significant effect on MSC osteogenic differentiation.

  • Both naïve and osteogenically induced MSCs exhibit higher osteogenic markers in reduced serum.

  • MSCs deposit the most calcium under 5 % O2 in osteogenic media supplemented with 5 % FBS.

  • Standard culture conditions (21 % O2, 10 % FBS) may not be optimal for MSC osteogenic differentiation.

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Acknowledgements

This project was supported by grants from the National Institutes of Health 1R03DE021704 and the Orthopaedic Research and Education Foundation (#13-006) to JKL and by the California Institute for Regenerative Medicine UC Davis Stem Cell Training Program (CIRM T1-00006, CIRM TG2-01163) to BYB.

Disclosures

The authors indicate no potential conflicts of interest.

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Authors and Affiliations

  1. Department of Biomedical Engineering, University of California, Davis, 451 Health Sciences Drive, Davis, CA, 95616, USA

    Bernard Y. K. Binder, John E. Sagun & J. Kent Leach

  2. Department of Orthopaedic Surgery School of Medicine, University of California, Davis, Sacramento, CA, 95817, USA

    J. Kent Leach

Authors
  1. Bernard Y. K. Binder
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  2. John E. Sagun
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  3. J. Kent Leach
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Correspondence to J. Kent Leach.

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Binder, B.Y.K., Sagun, J.E. & Leach, J.K. Reduced Serum and Hypoxic Culture Conditions Enhance the Osteogenic Potential of Human Mesenchymal Stem Cells. Stem Cell Rev and Rep 11, 387–393 (2015). https://doi.org/10.1007/s12015-014-9555-7

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  • DOI: https://doi.org/10.1007/s12015-014-9555-7

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