I have worked with the same type of cells before, but I never had that kind of a problem. I got that problem with cell lines, when I washed them twice with PBS or I let the PBS in contact for too long. That is due to the fact that PBS has no Ca in it, and the cells are very sensitive to Ca deprivation, because they need it to remain attached to the substrata. So the best way to wash is to add PBS, swirl the flask or vessel a couple of times and then remove it. In all it should not take more than 10 seconds.

That sounds rather strange; in my experience HASMC are very adhesive. In fact, I've had problems getting them to detach and disperse when passaging!

First, why are you feeding already confluent cell layers? In general it's best to passage just before full confluence. Perhaps the cells are overgrown, which can result in detaching in sheets. What kind of assay are you culturing the cells for?

Of course, I assume you're using TC plastics for your culture and not generic PS dishes?

Also, the above comment regarding calcium is keen; make sure your wash buffer contains Ca, if you are NOT trying to detach them. Before passaging, however, wash with Ca-free buffer.

Hi, I hope you sorted out cells coming off the flask problem. May I ask what media (and/or supplements) are you using? In our lab we will start working with human aortic smooth muscle cells and human aortic endothelial cells. I am trying to set up a protocol for culturing these cells. However, it seems a variety of different media and supplements are in use for these cells and I am confused which product to buy. Any help will be greatly appreciated! Thanks very much. 

Hi, can anyone who is working with HASMC can advice me the type of media they are using? I am trying to start the experiment with HASMC and there are different media, DMEM with different glucose concentration. Can someone shear the idea that works best for these cells.