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. 2006:2006:775-8.
doi: 10.1109/IEMBS.2006.259866.

Fibroblastic differentiation of human mesenchymal stem cells using connective tissue growth factor

Chang H Lee  1 , Eduardo K Moioli, Jeremy J Mao
Affiliations

Fibroblastic differentiation of human mesenchymal stem cells using connective tissue growth factor

Chang H Lee et al. Conf Proc IEEE Eng Med Biol Soc. 2006.

Abstract

The present study was designed to explore an ex vivo culturing protocol for fibroblastic differentiation of human mesenchymal stem cells (hMSCs) using connective tissue growth factor (CTGF). Fibroblastic differentiation from stem cells is of widespread significance in the engineering of virtually all tissues including tendons, ligaments, periodontal ligament, cranial sutures and as interstitial filler of all organs. The treatment with 100 ng/ml of recombinant human CTGF and 50 mug/ml ascorbic acids on monolayer cultured hMSCs showed significant increases in type I collagen and tenascin-C (Tn-C) contents by 2 and 4 wks. In addition, CTGF-treated hMSCs failed to show osteogenic or chondrogenic differentiation. The present data show that CTGF is an effective induction factor for fibroblastic differentiation of hMSCs. These findings have implications for engineering fibrous tissue by providing the initial evidence of a reproducible protocol for fibroblastic differentiation of hMSCs.

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Figures

Fig. 1
Fig. 1
Trichrome staining showing marked collagen synthesis upon CTGF for 4 wks. Treatment with 100 ng/ml of CTGF induced an increase in collagen deposition (scale=50 μm).
Fig. 2
Fig. 2
Collagen type I contents with or without CTGF treatment. Type I collagen contents in monolayer cultured hMSCs were significantly increased by the treatment with 100 ng/ml CTGF and 50 μg/ml ascorbic acid by 2 & 4 wks (n=3, *: p<0.01, **: p<0.001).
Fig. 3
Fig. 3
Tn-C contents with CTGF treatment. Tn-C contents, a maker for ligament fibroblasts, were significantly increased by the treatment with 100 ng/ml CTGF and 50 μg/ml ascorbic acid by 2 & 4wks (n=3, *: p<0.01, **: p<0.001).
Fig. 4
Fig. 4
Morphology of hMSCs with or without CTGF treatment. Both hMSCs and CTGF-treated hMSCs showed fibroblast-like spindle shape. However, there were no significant differences in cellular morphology caused by CTGF-treatment by 2 & 4 wks (scale = 200 μm).
Fig. 5
Fig. 5
Safranin O staining of hMSCs treated with CTGF (a, b) and chondrogenic medium (c) for 4 wks (scale = 100 μm).
Fig. 6
Fig. 6
von Kossa staining of hMSCs treated with CTGF (a, b) and osteogenic medium (c) for 4 wks (scale = 100 μm).
Fig. 7
Fig. 7
GAG and calcium deposition in hMSCs monolayer treated with CTGF or corresponding differentiation medium. GAG content and calcium deposition were not affected by the treatment with 100 ng/ml of CTGF by 2 & 4 wks (n=3, *: p<0.01, **: p<0.001).
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References

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