Perfusion affects the tissue developmental patterns of human mesenchymal stem cells in 3D scaffolds

Authors

  • Feng Zhao,

    1. Department of Chemical and Biomedical Engineering, Florida State University, Tallahassee, Florida
    Current affiliation:
    1. Department of Biomedical Engineering, Duke University, Durham, NC 27705.
    Search for more papers by this author
  • Warren L. Grayson,

    1. Department of Chemical and Biomedical Engineering, Florida State University, Tallahassee, Florida
    Current affiliation:
    1. Department of Biomedical Engineering, Columbia University, New York, NY 10027.
    Search for more papers by this author
  • Teng Ma,

    Corresponding author
    1. Department of Chemical and Biomedical Engineering, Florida State University, Tallahassee, Florida
    • Department of Chemical and Biomedical Engineering, Florida State University, 2525 Pottsdamer Street, Tallahassee, FL 32310.
    Search for more papers by this author
  • Andre Irsigler

    1. Department of Biological Science, Florida State University, Tallahassee, Florida
    Search for more papers by this author

Abstract

Human mesenchymal stem cells (hMSCs) developed in three-dimensional (3D) scaffolds are significantly affected by culture conditions. We hypothesized that the hydrodynamic forces generated in perfusion bioreactors significantly affected hMSC functionality in 3D scaffolds by shaping the extracellular matrix (ECM) proteins. In this study, hMSCs were grown in 3D poly(ethylene terephthalate) (PET) scaffolds in static and a parallel perfusion system under similar initial conditions for up to 35 days. Results demonstrated that even at very low media velocities (O [10−4 cm/sec]), perfusion cultures affected the ability of hMSCs to form an organized ECM network as illustrated by the immunostaining of collagen I and laminin fibrous structure. The change in the ECM microenvironment consequently influenced the nuclear shape. The hMSCs grown at the lower surface of static culture displayed a 15.2 times higher nuclear elongation than those at the upper surface, whereas cells grown in the perfusion bioreactor displayed uniform spherical nuclei on both surfaces. The difference in ECM organization and nuclear morphology associated with gene expression and differentiation characteristics of hMSCs. The cells exhibited lower CFU-F colony forming ability and decreased expressions of stem-cell genes of Rex-1 and Oct-4, implying a less primitive stem-cell phenotype was maintained in the perfusion culture relative to the static culture conditions. The significantly higher expression level of osteonectin gene in the perfusion culture at day 28 indicated an upregulation of osteogenic ability of hMSCs. The study highlights the critical role of dynamic culture conditions on 3D hMSC construct development and properties. J. Cell. Physiol. 219: 421–429, 2009. © 2009 Wiley-Liss, Inc.

Ancillary