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(Arteriosclerosis, Thrombosis, and Vascular Biology. 2006;26:1457.)
© 2006 American Heart Association, Inc.

Vascular Biology

New Insights to Vascular Smooth Muscle Cell and Pericyte Differentiation of Mouse Embryonic Stem Cells In Vitro

Henrik Lindskog; Elisabet Athley; Erik Larsson; Samuel Lundin; Mats Hellström; Per Lindahl

From the Institute of Biomedicine (H.L., E.L., P.L.), Department of Medical Biochemistry and Cell Biology, Sahlgrenska Academy, Göteborg University, Göteborg, Sweden; Angiogenetics Sweden AB (E.A., M.H.), Medicinaregatan 8A, Göteborg, Sweden; Institute of Biomedicine (S.L.), Department of Microbiology and Immunology, Sahlgrenska Academy, Göteborg University, Göteborg, Sweden.

Correspondence to Per Lindahl, Associate Professor, Wallenberg Laboratory of Cardiovascular Research, Bruna Stråket 16, Sahlgrenska University Hospital, SE-41345 Göteborg, Sweden. E-mail Per.Lindahl{at}wlab.gu.se

Objective— The molecular mechanisms that regulate pericyte differentiation are not well understood, partly because of the lack of well-characterized in vitro systems that model this process. In this article, we develop a mouse embryonic stem (ES) cell-based angiogenesis/vasculogenesis assay and characterize the system for vascular smooth muscle cell (VSMC) and pericyte differentiation.

Methods and Results— ES cells that were cultured for 5 days on OP9 stroma cells upregulated their transcription of VSMC and pericyte selective genes. Other SMC marker genes were induced at a later time point, which suggests that vascular SMC/pericyte genes are regulated by a separate mechanism. Moreover, sequence analysis failed to identify any conserved CArG elements in the vascular SMC and pericyte gene promoters, which indicates that serum response factor is not involved in their regulation. Gleevec, a tyrosine kinase inhibitor that blocks platelet-derived growth factor (PDGF) spell-receptor signaling, and a neutralizing antibody against transforming growth factor (TGF) ß1, ß2, and ß3 failed to inhibit the induction of vascular SMC/pericyte genes. Finally, ES-derived vascular sprouts recruited cocultured MEF cells to pericyte-typical locations. The recruited cells activated expression of a VSMC- and pericyte-specific reporter gene.

Conclusions— We conclude that OP9 stroma cells induce pericyte differentiation of cocultured mouse ES cells. The induction of pericyte marker genes is temporally separated from the induction of SMC genes and does not require platelet-derived growth factor B or TGFß1 signaling.

We developed an ES cell-based angiogenesis/vasculogenesis model and characterized the system for VSMC and pericyte differentiation. Five days of culture on OP9 feeder cells induced expression of VSMC/pericyte-specific genes in mouse ES cells. The induction was temporally separated from the induction of SMC-specific genes, which suggest regulation by separate mechanisms. Finally, PDGFB and TGFß1 were dispensable for pericyte differentiation.

Key Words: angiogenesis • embryonic stem cells • pericytes • vascular smooth muscle cells • vasculogenesis

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