Adenoviral-mediated transfer of TGF-β1 but not IGF-1 induces chondrogenic differentiation of human mesenchymal stem cells in pellet cultures

Abstract

Objective. The objective of the present study was to investigate the potential of application of growth factor genes to induce chondrogenic differentiation of human-derived mesenchymal stem cells (MSCs). The growth factor genes evaluated in the present study were transforming growth factor 1 (TGF-β1) and insulin-like growth factor 1 (IGF-1).Methods. Human MSCs were transduced with the adenoviral vectors carrying either TGF-β1 or IGF-1 (AdTGF-β1 and AdIGF-1 respectively) or a combination of both growth factor genes at different multiplicities of infection (MOI) and were then made into pellets. Pellets were also made from nontransduced cells and maintained in culture medium supplemented with 10 ng/mL of TGF-β1. At specified time points, histological analysis, cartilage matrix gene expression, and immunofluorescence were performed to determine the extent of chondrogenic differentiation.Results. MSCs transduced with the AdTGF-β1 demonstrated robust chondrogenic differentiation, while those made from AdIGF-1 did not. AdTGF-β1 pellets demonstrated aggrecan gene expression as early as day 3 of pellet culture, while type II collagen gene expression was detected by day 10 of culture. The AdIGF-1, alone or in combination with TGF-β1 pellets, did not show any type II collagen gene expression at any time point. By immunofluoresecence, type X collagen was distributed throughout the matrix in TGF-β1 protein pellets while the growth factor gene pellets displayed scant staining. Conclusion.The results suggest that sustained administration of TGF-β1 may be more effective in suppressing terminal differentiation than intermittent dosing and thus effective for cartilage repair.

Adeno-associated viral gene transfer of transforming growth factor-β1 to human mesenchymal stem cells improves cartilage repair

“…[15][16][17] The chondrogenic potential of human mesenchymal stem cells (hMSC) is well described. 18,19 Addition of transforming growth factor-beta-1 (TGF-b1) to hMSC has been shown to consistently induce chondrogenesis while avoiding terminal differentiation [20][21][22][23][24] Adenoviral mediated transfer of TGF-b1 genes to hMSC has been shown to similarly induce chondrogenesis and inhibit terminal differentiation. 21 Efficient and sustained gene delivery to mesenchymal stem cells with adenovirus, lentivirus and retrovirus have been described.…”

“…18,19 Addition of transforming growth factor-beta-1 (TGF-b1) to hMSC has been shown to consistently induce chondrogenesis while avoiding terminal differentiation [20][21][22][23][24] Adenoviral mediated transfer of TGF-b1 genes to hMSC has been shown to similarly induce chondrogenesis and inhibit terminal differentiation. 21 Efficient and sustained gene delivery to mesenchymal stem cells with adenovirus, lentivirus and retrovirus have been described. 25 Although viral vectors are effective agents for gene transfer, there are significant safety concerns regarding the use of adenoviruses and retroviruses for clinical gene therapy.…”

Periosteal Cell Pellet Culture System: A New Technique for Bone Engineering

“…After cells were dehydrated Pellet culture systems are established methods of culturing chondrocytes and mesenchymal stem cells for through graded concentrations of ethanol alone, a mixture of MNA (methyl nadic anhydride), Epon 812 Resin chondrogenesis (5,8,22,25,37,38). Lee et al (25) compared a new pellet culture system for human interverte-(TAAB, Berkshire, UK), DDSA (dodecenyl succinic anhydride), and DMP30 [2,4,6-tri (dimethylaminobral disc cells to a conventional alginate bead system in culture.…”

Chondrogenic differentiation of adipose tissue‐derived mesenchymal stem cells: Greater doses of growth factor are necessary

“…Based on previous studies which showed that IGF alone did not induce chondrogenesis from BMMSC, 28,29 we did not study the effects of increasing dose of IGF-I alone in this study. Instead, we added IGF-I in the same scale to the increased doses of TGF-b 2 .…”