Enhanced in vitro chondrogenesis of primary mesenchymal stem cells by combined gene transfer

Andre F. Steinert, Glyn D. Palmer, Carmencita Pilapil, Ulrich Nöth, Christopher H Evans, Steven C. Ghivizzani

Research output: Contribution to journalArticle

78 Citations (Scopus)

Abstract

Because articular cartilage has a poor regeneration capacity, numerous cell-based approaches to therapy are currently being explored. The present study involved the use of gene transfer as a means to provide sustained delivery of chondrogenic proteins to primary mesenchymal stem cells (MSCs). In previous work, we found that adenoviral-mediated gene transfer of transforming growth factor-beta1 (TGF-β1) and bone morphogenetic protein 2 (BMP-2), but not insulin-like growth factor 1 (IGF-1), could be used to induce chondrogenic differentiation of MSCs in an aggregate culture system. In the present study, we examined the effects on chondrogenesis of these transgenes when delivered in combination. Cultures of bone marrow-derived MSCs were infected with 2.5 × 102 or 2.5 × 103 viral particles/cell of each adenoviral vector individually, or in combination, seeded into aggregates, and cultured for 3 weeks in a defined serum-free medium. Levels of transgene product in the medium were initially high, approximately 100 ng/mL TGF-β1, 120 ng/mL BMP-2, and 80 ng/mL IGF-1 at day 3, and declined thereafter. We found that co-expression of IGF-1 and TGF-β1, BMP-2, or both at low doses resulted in larger aggregates, higher levels of glycosaminoglycan synthesis, stronger staining for proteoglycans and collagen type II and X, and greater expression of cartilage-specific marker genes than with either transgene alone. Gene-induced chondrogenesis of MSCs using multiple genes that act synergistically may enable the administration of reduced viral doses in vivo and could be of considerable benefit for the development of cell-based therapies for cartilage repair.

Original languageEnglish (US)
Pages (from-to)1127-1139
Number of pages13
JournalTissue Engineering - Part A
Volume15
Issue number5
DOIs
StatePublished - May 1 2009
Externally publishedYes

Fingerprint

Gene transfer
Chondrogenesis
Stem cells
Mesenchymal Stromal Cells
Transforming Growth Factor beta1
Bone Morphogenetic Protein 2
Cartilage
Somatomedins
Transgenes
Genes
Insulin
Bone
Cell culture
Proteins
Collagen Type X
Collagen Type II
Serum-Free Culture Media
Proteoglycans
Glycosaminoglycans
Articular Cartilage

ASJC Scopus subject areas

  • Bioengineering
  • Biochemistry
  • Biomedical Engineering
  • Biomaterials

Cite this

Enhanced in vitro chondrogenesis of primary mesenchymal stem cells by combined gene transfer. / Steinert, Andre F.; Palmer, Glyn D.; Pilapil, Carmencita; Nöth, Ulrich; Evans, Christopher H; Ghivizzani, Steven C.

In: Tissue Engineering - Part A, Vol. 15, No. 5, 01.05.2009, p. 1127-1139.

Research output: Contribution to journalArticle

Steinert, Andre F. ; Palmer, Glyn D. ; Pilapil, Carmencita ; Nöth, Ulrich ; Evans, Christopher H ; Ghivizzani, Steven C. / Enhanced in vitro chondrogenesis of primary mesenchymal stem cells by combined gene transfer. In: Tissue Engineering - Part A. 2009 ; Vol. 15, No. 5. pp. 1127-1139.
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