Derivation of contractile smooth muscle cells from embryonic stem cells.
Smooth muscle cells (SMCs) play a key role in vascular physiology and pathology. An appreciation of normal SMCs developmental mechanisms will likely lead to a better understanding of disease processes and potentially to novel treatment strategies. We present a method for generating relatively pure populations of SMCs from embryonic stem cells (ESC) which display appropriate excitation and contractile responses to vasoactive agonists. We also present protocols for assessment of SMCs purity and identity by immunofluorescence, quantitative RT-PCR, and FACS. This ESC-based system has tremendous potential for studying developmental regulation of SMC lineage, as well as for possible SMC tissue engineering.
Sinha
Sanjay
S
Division of Cardiovascular Medicine, University of Cambridge, Addenbrooke's Hospital, Cambridge, UK.
Hoofnagle
Mark H
MH
Owens
Gary K
GK
D000199:Actins / Q000378:metabolism; D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation; D002460:Cell Line; D002469:Cell Separation; D004622:Embryo, Mammalian / Q000166:cytology; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism; D019332:Endothelin-1 / Q000378:metabolism; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D020869:Gene Expression Profiling; D018507:Gene Expression Regulation, Developmental; D051379:Mice; D008858:Microscopy, Phase-Contrast; D009119:Muscle Contraction; D032389:Myocytes, Smooth Muscle / Q000166:cytology* / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction
D000199:Actins; D015415:Biomarkers; D019332:Endothelin-1