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Efficient generation of smooth muscle cells from adipose‐derived stromal cells by 3D mechanical stimulation can substitute the use of growth factors in vascular tissue engineering
Authors:Mojtaba Parvizi  Lydia AM Bolhuis‐Versteeg  André A Poot  Martin C Harmsen
Institution:1. University of Groningen, University Medical Center Groningen, Department of Pathology and Medical Biology, Groningen, the Netherlands;2. Biomaterials Science and Technology, University of Twente, Enschede, the Netherlands
Abstract:Occluding artery disease causes a high demand for bioartificial replacement vessels. We investigated the combined use of biodegradable and creep‐free poly (1,3‐trimethylene carbonate) (PTMC) with smooth muscle cells (SMC) derived by biochemical or mechanical stimulation of adipose tissue‐derived stromal cells (ASC) to engineer bioartificial arteries. Biochemical induction of cultured ASC to SMC was done with TGF‐β1 for 7d. Phenotype and function were assessed by qRT‐PCR, immunodetection and collagen contraction assays. The influence of mechanical stimulation on non‐differentiated and pre‐differentiated ASC, loaded in porous tubular PTMC scaffolds, was assessed after culturing under pulsatile flow for 14d. Assays included qRT‐PCR, production of extracellular matrix and scanning electron microscopy. ASC adhesion and TGF‐β1‐driven differentiation to contractile SMC on PTMC did not differ from tissue culture polystyrene controls. Mesenchymal and SMC markers were increased compared to controls. Interestingly, pre‐differentiated ASC had only marginal higher contractility than controls. Moreover, in 3D PTMC scaffolds, mechanical stimulation yielded well‐aligned ASC‐derived SMC which deposited ECM. Under the same conditions, pre‐differentiated ASC‐derived SMC maintained their SMC phenotype. Our results show that mechanical stimulation can replace TGF‐β1 pre‐stimulation to generate SMC from ASC and that pre‐differentiated ASC keep their SMC phenotype with increased expression of SMC markers.
Keywords:Adipose tissue‐derived stromal cells (ASC)  Poly(1  3‐trimethylene carbonate) (PTMC)  Smooth muscle cells (SMC)  Transforming Growth Factor beta 1 (TGF‐β  1)  Vascular tissue engineering
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