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

Feng Zhao, Warren L Grayson, Teng Ma, Andre Irsigler

Research output: Contribution to journalArticle

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.

Original languageEnglish (US)
Pages (from-to)421-429
Number of pages9
JournalJournal of Cellular Physiology
Volume219
Issue number2
DOIs
StatePublished - May 2009
Externally publishedYes

Fingerprint

Stem cells
Mesenchymal Stromal Cells
Scaffolds
Perfusion
Tissue
Cell culture
Extracellular Matrix
Bioreactors
pX Genes
Stem Cells
Osteonectin
Genes
Polyethylene Terephthalates
Extracellular Matrix Proteins
Laminin
Hydrodynamics
Up-Regulation
Collagen
Gene expression
Elongation

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Cell Biology
  • Physiology

Cite this

Perfusion affects the tissue developmental patterns of human mesenchymal stem cells in 3D scaffolds. / Zhao, Feng; Grayson, Warren L; Ma, Teng; Irsigler, Andre.

In: Journal of Cellular Physiology, Vol. 219, No. 2, 05.2009, p. 421-429.

Research output: Contribution to journalArticle

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