Sinha Sanjay, Hoofnagle Mark H, Owens Gary K
Division of Cardiovascular Medicine, University of Cambridge, Addenbrooke's Hospital, Cambridge, UK.
Methods Mol Biol. 2009;482:345-67. doi: 10.1007/978-1-59745-060-7_22.
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.
平滑肌细胞（SMCs）在血管生理和病理过程中发挥着关键作用。深入了解正常平滑肌细胞的发育机制可能有助于更好地理解疾病过程，并有可能带来新的治疗策略。我们提出了一种从胚胎干细胞（ESC）中生成相对纯净平滑肌细胞群体的方法，这些细胞对血管活性激动剂表现出适当的兴奋和收缩反应。我们还提出了通过免疫荧光、定量逆转录-聚合酶链反应（RT-PCR）和荧光激活细胞分选（FACS）评估平滑肌细胞纯度和特性的方案。这种基于胚胎干细胞的系统在研究平滑肌细胞谱系的发育调控以及可能的平滑肌组织工程方面具有巨大潜力。
