Spiekman, M. (Maroesjka) (2012) Human adipose tissue-derived stromal cells suppress TGF-β induced fibroblast differentiation of human dermal fibroblasts: implications for scar formation. thesis, Medicine.
Full text available on request.Abstract
Adipose tissue-derived stromal cells (ADSC) are multipotent stem cells which can be isolated and cultured from white adipose tissues throughout the body. In wound healing and regeneration of the skin, ADSC have already proven to be effective. However, it is not yet known if and how ADSC can prevent or reduce the formation of dermal scars. Scar formation is caused by persisting stimulation of fibroblasts by Transforming Growth Factor- β (TGF-β), which causes them to differentiate to myofibroblasts. The aim of this research is to investigate the influence of ADSC on TGF-β-mediated fibroblast-myofibroblast differentiation. We hypothesize that ADSC inhibit fibrotic remodeling by (1) repressing myofibroblast formation and (2) increasing matrix turnover. ADSC were isolated from human adipose tissue. ADSC conditioned medium (ADSC CM) was obtained by incubating ADSC with culture medium for 24h, thus collecting trophic factors from ADSC. Human dermal fibroblasts, adult type (HDFa) were incubated with ADSC CM or co-cultured with ADSC, in presence or absence of TGF-β1. After 4 days, HDFa were collected for gene transcript and immunoblot analysis of collagen I and III, matrix metalloproteinases (MMP), tissue inhibitors of metalloproteinases (TIMP) and SM22α, a mesenchymal marker. Functionally, contraction of HDFa was measured by means of a gel contraction assay. HDFa proliferation is up regulated by TGF-β1. ADSC CM brings proliferation back to control levels. ADSC CM reduces SM22α gene and protein expression in HDFa to 0.38±0.02 and 0.45±0.09 fold, respectively. After TGF-β1 stimulation, SM22α protein increased to 1.71±0.19 fold which was brought back to 1.11±0.16 fold by ADSC CM. Collagen I and collagen III mRNA expression to 0.49± 0.04 fold and 0.48± 0.06 fold. On protein level, collagen I is reduced to 0.51±0.13 fold. Furthermore, MMP-1, MMP-2, MMP-14 and TIMP-1 and TIMP-2 gene expression in HDFa was up regulated by ADSC CM or direct co-culture with ADSC. Collagen gel contraction by HDFa was significantly reduced after ADSC CM, irrespective of TGF-β1 stimulation. In this study we show ADSC inhibit fibrotic remodeling by repressing myofibroblast formation and by increasing matrix remodeling. In term, inhibition of fibroblast differentiation might lead to prevention or even reduction of dermal scars.
| Item Type: | Thesis (Thesis) |
|---|---|
| Supervisor name: | Under supervision of: and Harmsen, M.C. and Cardiovascular Regenerative Medicine group and Department of Pathology and Medical Biology and University Medical Centre Groningen |
| Faculty: | Medical Sciences |
| Date Deposited: | 25 Jun 2020 10:54 |
| Last Modified: | 25 Jun 2020 10:54 |
| URI: | https://umcg.studenttheses.ub.rug.nl/id/eprint/1520 |
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