Environmental parameters influence non-viral transfection of human mesenchymal stem cells for tissue engineering applications

William J. King, Nicholas A. Kouris, Siyoung Choi, Brenda M. Ogle, William L. Murphy

Research output: Contribution to journalArticlepeer-review

25 Scopus citations

Abstract

Non-viral transfection is a promising technique that could be used to increase the therapeutic potential of stem cells. The purpose of this study was to explore practical culture parameters of relevance in potential human mesenchymal stem cell (hMSC) clinical and tissue engineering applications, including type of polycationic transfection reagent, N/P ratio and dose of polycation/pDNA polyplexes, cell passage number, cell density and cell proliferation. The non-viral transfection efficiency was significantly influenced by N/P ratio, polyplex dose, cell density and cell passage number. hMSC culture conditions that inhibited cell division also decreased transfection efficiency, suggesting that strategies to promote hMSC proliferation may be useful to enhance transfection efficiency in future tissue engineering studies. Non-viral transfection treatments influenced hMSC phenotype, including the expression level of the hMSC marker CD105 and the ability of hMSCs to differentiate down the osteogenic and adipogenic lineages. The parameters found here to promote hMSC transfection efficiency, minimize toxicity and influence hMSC phenotype may be instructive in future non-viral transfection studies and tissue engineering applications.

Original languageEnglish (US)
Pages (from-to)689-699
Number of pages11
JournalCell and Tissue Research
Volume347
Issue number3
DOIs
StatePublished - Mar 2012

Bibliographical note

Funding Information:
Acknowledgments The authors are grateful for support from the AO Research Foundation (GENEDEL exploratory research grant to William L. Murphy) and the National Science Foundation (Graduate Research Fellowship to William J. King and CAREER award #0745563 to William L. Murphy)

Copyright:
Copyright 2012 Elsevier B.V., All rights reserved.

Keywords

  • Adult stem cells
  • Bioprocessing
  • Multipotency
  • Polyethyleneimine
  • Toxicity

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