Background Cell source is one of the most important constructions for tissue engineered blood vessels(TEBV). As human adult vascular cells are limited by the replicative life spans and poor collagen secretion, stem ce...Background Cell source is one of the most important constructions for tissue engineered blood vessels(TEBV). As human adult vascular cells are limited by the replicative life spans and poor collagen secretion, stem cell has become a promising cell source. Hence, we investigated the differentiation of human induced pluripotent stem cells(hiPSC) into functional smooth-muscle-like cells(SMLCs) by embryoid bodies method and explored whether transforming growth factor-β1(TGF-β1) can promote the differentiation. Methods HiPSCs were cultured in smooth muscle cell medium with or without TGF-β1 after forming embryoid bodies. The cell morphology, cell characteristics and contractility were compared after 7 days of differentiation. Real-time PCR and Western blot were used to assess the mRNA and protein expression levels of α-SMA, Calponin, SM22α, Collagen I and Collagen III. Functional contraction study was performed using carbachol. Results HiPSC could successfully differentiate into cells that were similar to typical smooth muscle cells in morphology. The expression of α-SMA, Calponin and SM22α up-regulated after induction. TGF-β1 could further up-regulated α-SMA expression.Immunofluorescence images showed that more than 80% of the hiPSC-derived SMLCs by TGF-β1 stained with smooth muscle cell markers α-SMA, SMMHC, SM22α and Calponin. Analyses of expression in collagen showed that hiPSC-derived SMLCs exhibited higher levels of Collagen I and Collagen III after induction by TGF-β1. Conclusion The hiPSC could successfully differentiate into smooth-muscle-like cells using embryoid bodies method. TGF-β1 can promote the differentiation and enhance collagen synthesis[.S Chin J Cardiol 2019;20(1):44-53]展开更多
基金supported by Science and Technology Planning Project of Guangdong Province,China(No.2016B070701007)
文摘Background Cell source is one of the most important constructions for tissue engineered blood vessels(TEBV). As human adult vascular cells are limited by the replicative life spans and poor collagen secretion, stem cell has become a promising cell source. Hence, we investigated the differentiation of human induced pluripotent stem cells(hiPSC) into functional smooth-muscle-like cells(SMLCs) by embryoid bodies method and explored whether transforming growth factor-β1(TGF-β1) can promote the differentiation. Methods HiPSCs were cultured in smooth muscle cell medium with or without TGF-β1 after forming embryoid bodies. The cell morphology, cell characteristics and contractility were compared after 7 days of differentiation. Real-time PCR and Western blot were used to assess the mRNA and protein expression levels of α-SMA, Calponin, SM22α, Collagen I and Collagen III. Functional contraction study was performed using carbachol. Results HiPSC could successfully differentiate into cells that were similar to typical smooth muscle cells in morphology. The expression of α-SMA, Calponin and SM22α up-regulated after induction. TGF-β1 could further up-regulated α-SMA expression.Immunofluorescence images showed that more than 80% of the hiPSC-derived SMLCs by TGF-β1 stained with smooth muscle cell markers α-SMA, SMMHC, SM22α and Calponin. Analyses of expression in collagen showed that hiPSC-derived SMLCs exhibited higher levels of Collagen I and Collagen III after induction by TGF-β1. Conclusion The hiPSC could successfully differentiate into smooth-muscle-like cells using embryoid bodies method. TGF-β1 can promote the differentiation and enhance collagen synthesis[.S Chin J Cardiol 2019;20(1):44-53]
