Heterogeneity of embryological origins is a hallmark of vascular smooth muscle cells (SMCs) and may influence the development of vascular disease. Differentiation of human pluripotent stem cells (hPSCs) into developmental origin–specific SMC subtypes remains elusive. Here we describe a chemically defined protocol in which hPSCs were initially induced to form neuroectoderm, lateral plate mesoderm or paraxial mesoderm. These intermediate populations were further differentiated toward SMCs (>80% MYH11+ and ACTA2+), which displayed contractile ability in response to vasoconstrictors and invested perivascular regions in vivo. Derived SMC subtypes recapitulated the unique proliferative and secretory responses to cytokines previously documented in studies using aortic SMCs of distinct origins. Notably, this system predicted increased extracellular matrix degradation by SMCs derived from lateral plate mesoderm, which was confirmed using rat aortic SMCs from corresponding origins. This differentiation approach will have broad applications in modeling origin-dependent disease susceptibility and in developing bioengineered vascular grafts for regenerative medicine.
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The authors thank L. Vallier and S.T. Rashid for supplying us the wild-type human iPSCs; K. Jensen for the human fetal gut RNA; N. Figg and M. Ackers-Johnson for help rendered in Matrigel sectioning and the harvesting of rat aortic SMCs, respectively. We also thank T. Faial and D. Ortmann for validation of the mesoderm protocols. This work was supported by a Wellcome Trust Intermediate Clinical Fellowship for S.S. and the Cambridge National Institute for Health Research Comprehensive Biomedical Research Centre. C. Cheung was sponsored by a National Science Scholarship (PhD) from the Agency for Science, Technology and Research (Singapore). R.A.P. and M.W.B.T. were supported by a Medical Research Council centre grant and A.S.B. was supported by a Leukemia and Lymphoma Society grant.
The authors declare no competing financial interests.
Supplementary Tables 1–3 and Supplementary Figs. 1–6 (PDF 1511 kb)
HeLa cells (WMV 343 kb)
NE-SMC (WMV 142 kb)
LM-SMC (WMV 230 kb)
PM-SMC (WMV 198 kb)
ASMC (WMV 150 kb)
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Cheung, C., Bernardo, A., Trotter, M. et al. Generation of human vascular smooth muscle subtypes provides insight into embryological origin–dependent disease susceptibility. Nat Biotechnol 30, 165–173 (2012). https://doi.org/10.1038/nbt.2107
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