Efficient lentiviral transduction of human mesenchymal stem cells that preserves proliferation and differentiation capabilities

Paul Lin, Yuan Lin, Donald P. Lennon, Diego Correa, Mark Schluchter, Arnold I. Caplan

Research output: Contribution to journalArticle

40 Scopus citations

Abstract

Long-term lentiviral transduction of human mesenchymal stem cells (hMSCs) greatly enhances the usefulness of these cells. However, such transduction currently requires the use of polybrene, which severely inhibits hMSC proliferation. In contrast, protamine sulfate at 100 g/ml doubled transduction efficiencies without affecting proliferation or differentiation potential. Expression levels improved 2.2-fold with the addition of a woodchuck hepatitis post-transcriptional regulatory element. Further improvements in transduction efficiencies could be obtained by a modest increase in viral concentrations through increased viral titers or decreased transduction volumes without changing multiplicity of infection, by transducing over multiple days, or by culturing the cells in fibroblast growth factor-2. Centrifugation improved expression but had no effect on efficiency. Transgene expression was stable over 6 weeks in vitro and in vivo. Donor-to-donor and intradonor variability were observed in primary passage through passage 2 cultures, but not at passage 3. These results provide a better optimized approach for expanded use of hMSCs through genetic manipulation.

Original languageEnglish (US)
Pages (from-to)886-897
Number of pages12
JournalStem Cells Translational Medicine
Volume1
Issue number12
DOIs
StatePublished - Jan 1 2012
Externally publishedYes

Keywords

  • Adult human bone marrow
  • Adult stem cells
  • Cellular therapy
  • Gene therapy
  • Lentiviral vector
  • Mesenchymal stem cells
  • Osteoblast
  • Transduction

ASJC Scopus subject areas

  • Developmental Biology
  • Cell Biology

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