Transforming growth factor-beta-induced transition of fibroblasts: a model for myofibroblast procurement in tissue valve engineering.

摘要: Background and aim of the study The selection a suitable cell type for scaffold seeding, its isolation adequate expansion in vitro remains major challenge tissue valve engineering. was to establish model efficient procurement myofibroblasts in-vitro seeding using fibroblasts as progenitor cells. Methods Dermal arterial mesenchymal cells from human (hDMC1.1 hAMC1.1) sheep (sDMC1.1 sAMC1.1) were isolated by sequential culture. Cell isolates characterized stringent criteria based on morphology, immunocytochemistry antibodies vimentin, cytokeratin, prolyl 4-hydroxylase, smooth muscle alpha-actin (alpha-SMA) myosin, Western blotting alpha-SMA N-cadherin. Cultures with less than 10-20% alpha-SMA-positive considered be fibroblastic. Cells subsequently transdifferentiated cytokine transforming growth factor-beta1 (TGF-beta1) during five days, then evaluated morphotypically, immunocytochemistry, blotting. metabolic functional properties TGF-beta1-treated untreated colonies compared measuring expression extracellular proteins (collagen 1 tenascin-C) collagen matrix contraction assay. Results TGF-beta1 successfully transformed both metabolically active myofibroblast characterization. Alpha-SMA positivity 100% obtained all cases (hDMC1.1, hAMC1.1, sDMC1.1, after transformation 50% non-transformed state (hAMC1.1, 17%; hDMC1.1, 10%; sAMC1.1, 43%; 30%). This observation further supported increased contractility an up-regulation protein production Conclusion Untreated were, at best, alpha-SMA-positive. By allowing high densities relatively short time, seen potentially useful tool engineering, least investigations autologous model.