Porcine bone marrow‐derived mesenchymal stem cells as cell source for smooth muscle cells in small diameter vascular tissue engineered grafts

Abstract

Preconditioning of a synthetic graft with autologous cells is considered the best option to obtain functional vascular grafts. These cells are preferably isolated from an easy accessible patient‐autologous source, such as bone marrow or blood. We investigated if bone marrow‐derived mesenchymal stem cells (MSCs) can be isolated, cultured and instructed to gain a vascular smooth muscle cell (SMC) phenotype.MSCs were isolated from porcine bone marrow by gradient centrifugation and cultured in MSC or SMC‐defined culture media. Cells were analysed for SMC phenotype by determination of mRNA and protein expression of alpha‐SMA, calponin, desmin and collagen.Porcine MSCs can be reproducibly isolated and cultured. No eminent differences in growth dynamics (doubling time, morphology) were observed between animals. Cells grew in multiple layers and showed mesenchymal morphology with an elongated cell shape. Induction of expression of all SMC markers, both at mRNA and protein level, was observed and was highest in cells cultured in MSC‐defined medium.We conclude that porcine MSCs can be reproducibly isolated and differentiated into cells with SMC‐phenotype with sufficient efficacy. We show that the current protocol allows the MSCs to be readily used for application in small diameter vascular grafts.