2016
DOI: 10.3791/53983
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Isolation of Murine Coronary Vascular Smooth Muscle Cells

Abstract: 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 CaCl 2 . The perfusates were collected at 15 min intervals for 90 min, pelleted by centrifugation, resuspended in plating media, an… Show more

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Cited by 4 publications
(3 citation statements)
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“…Magnified FOVs showing immunostained CAFs and the image segmentation used to identify the fibroblasts are presented in Figures 5B and 5C for the MDA-MB-231 tumor section, and in Figures 5E and 5F for the Clone 13 tumor section. Since smooth muscle cells also express α-SMA [ 28 ], vessel regions were excluded in the analysis. Quantification of immunostaining identified higher CAFs in MDA-MB-231 tumors compared to Clone 13 tumors, as shown in Figure 5G .…”
Section: Resultsmentioning
confidence: 99%
“…Magnified FOVs showing immunostained CAFs and the image segmentation used to identify the fibroblasts are presented in Figures 5B and 5C for the MDA-MB-231 tumor section, and in Figures 5E and 5F for the Clone 13 tumor section. Since smooth muscle cells also express α-SMA [ 28 ], vessel regions were excluded in the analysis. Quantification of immunostaining identified higher CAFs in MDA-MB-231 tumors compared to Clone 13 tumors, as shown in Figure 5G .…”
Section: Resultsmentioning
confidence: 99%
“…Primary mouse VSMCs were isolated from aortae of 8‐week‐old mice (from the aortic roots to bifurcation of the renal arteries) as described 33 . VSMCs were cultured in complete Dulbecco's modified Eagle's medium at 37°C in an atmosphere containing 5% CO 2 .…”
Section: Methodsmentioning
confidence: 99%
“…Additionally, THP-1 cells were supplemented with 162 nM of phorbol myristate acetate (PMA) (524400, Sigma-Aldrich, St. Louis, MO, USA) for 48 h to stimulate terminal differentiation. Primary rat aortic smooth muscle cells (RASMC) from Sprague Dawley rats (10–12 weeks old), were isolated from thoracic aortae as previously described using an explant method [ 23 ]. RASMC were used between passages 4–9 for all experiments.…”
Section: Methodsmentioning
confidence: 99%