Isolation of Murine Coronary Vascular Smooth Muscle Cells.
While the isolation and culture of vascular smooth muscle cells (VSMCs) from large vessels is well established, we sought to isolate and culture VSMCs from the coronary circulation. Hearts with intact aortic arches were removed and perfused via retrograde Langendorff with digestion solution containing 300 Units/ml of collagenase type II, 0.1 mg/ml soybean trypsin inhibitor and 1 M CaCl2. The perfusates were collected at 15 min intervals for 90 min, pelleted by centrifugation, resuspended in plating media, and plated on tissue culture dishes. VSMCs were characterized by presence of SM22α, α-SMA, and vimentin. One of the main advantages of using this technique is the ability to isolate VSMCs from the coronary circulation of mice. Although the small number of cells obtained can limit some of the applications for which the cells can be utilized, isolated coronary VSMCs can be used in a variety of well-established cell culture techniques and assays. Studies investigating VSMCs from genetically modified mice can provide further information about structure-function and signaling processes associated with vascular pathologies.
School of Biomedical Science, The Ohio State University College of Medicine; Center for Cardiovascular and Pulmonary Research, The Research Institute at Nationwide Children's Hospital.
Kathryn E
KE
Husarek
Center for Cardiovascular and Pulmonary Research, The Research Institute at Nationwide Children's Hospital.
Xiaojin
X
Zhang
Center for Cardiovascular and Pulmonary Research, The Research Institute at Nationwide Children's Hospital.
Patricia E
PE
McCallinhart
Center for Cardiovascular and Pulmonary Research, The Research Institute at Nationwide Children's Hospital; Department of Pediatrics, The Ohio State University College of Medicine.
Pamela A
PA
Lucchesi
Center for Cardiovascular and Pulmonary Research, The Research Institute at Nationwide Children's Hospital; Department of Pediatrics, The Ohio State University College of Medicine; aaron.trask@nationwidechildrens.org.
Aaron J
AJ
Trask
D000818:Animals; D018929:Cell Culture Techniques; D002469:Cell Separation; D002478:Cells, Cultured; D003331:Coronary Vessels / Q000166:cytology*; D051379:Mice; D009131:Muscle, Smooth, Vascular / Q000166:cytology; D032389:Myocytes, Smooth Muscle