most recent 30 from http://asksci.com 2010-07-29T19:14:06Z http://asksci.com/feeds/question/145 http://www.creativecommons.org/licenses/by-nc/2.5/rdf http://asksci.com/questions/145/primary-smooth-muscle-cell-culture Kyle Legate 2010-02-03T14:22:59Z 2010-02-03T14:22:59Z

<p>I am trying to do some functional assays on primary smooth muscle cells from young mice. When I say young I mean newborn esophagus or 3 week old aortic smooth muscle. There are not many cells for functional assays so I take them into cell culture for a while until they grow confluent, and then I try to stimulate them with an agonist (carbachol) and visualize contraction under the microscope. The problem appears to be that these cells lose their contractility really fast and I can't see any response to agonist, although smooth muscle actin expression is still quite high (so they are still 'smooth muscle cells'). I culture them in DMEM + 10% FBS on plastic. I read that primary SMCs will rexpress contractility makers if starved for 3 days, but this hasn't helped my situation--maybe this trick is human-specific?</p> <p>Does anyone have any tips on how I can keep these cells contraction-competent in culture?</p>