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Recent studies suggested that the post-natal mesothelium retain differentiative potential of the embryonic mesothelium, which generates fibroblasts and vascular smooth muscle cells (VSMCs), in developing coelomic organs via epithelial-to-mesenchymal transition (EMT). Whether adult mesothelial cells (MCs) are able to give rise to functional VSMCs in vitro and which are the factors and mechanisms directing this process remain largely unknown. Here, we isolated adipose tissue MCs (ATMCs) from adult mice, and demonstrated that ATMCs cultured in a serum-containing media supplemented with epidermal growth factor (EGF) efficiently increased both their proliferation and EMT above levels found in only serum-containing media cultures. EGF-induced ATMCs gained phosphorylation of the EGF receptor and activated simultaneously ILK/Erk1/2, PI3K/Akt and Smad2/3-dependent pathways. Sequential subculture onto collagen-I surface efficiently improved their vasculogenic EMT towards cells featuring VSMCs (α-SMA, calponin, caldesmon, SM22α, desmin, SM-MHC, smoothelin-B and PDGFR-β) that could actively contract in response to receptor and non-receptor-mediated vasoactive agonists. Overall, our results indentify EGF signalling as a robust vasculogenic inductive pathway for ATMCs, leading to their transdifferentiation into functional VSMC-like cells.The authors are supported by the Fundación\udProgreso y Salud, Consejería de Salud, Junta de Andalucía (Grant PI-0022/2008);\udFEDER co-funded grants from Consejería de Innovación Ciencia y Empresa, Junta\udde Andalucía (Grant CTS-6505; INP-2011-1615-9); FEDER co-funded grants\udfrom Instituto de Salud Carlos III (Red TerCel-Grant RD12/0019/0028; PI10/00964\udand PI10/00871) and the Ministry of Health and Consumer Affairs (Advanced\udTherapies Program Grant TRA-120). Support from FSED and FAID allow access to\uddatabanks. CIBERDEM is an initiative of the Instituto de Salud Carlos IIIPeer Reviewe

EGF-induced adipose tissue mesothelial cells undergo functional vascular smooth muscle differentiation