Primaria/gelatin plates and gravity-defying fixed cells - (Mar/08/2010 )
I am culturing primary neonatal rat ventricular myocytes (NRVMs) to use for ChIP experiments. The cells are grown on 100 mm BD Primaria dishes pre-coated with gelatin. To harvest the cells for ChIP, they must first be fixed in formaldehyde and then scraped from the plate in PBS and collected.
The problem I am having is during the cell scraping, the cells love to 'float' up the dish, defying gravity and working their way up any residual liquid across the surface of the dish. I don't know if this is specifically due to charge from the Primaria surface treatment or some other reason. This problem makes it very difficult to collect all the cells, even after three or four sequential rinses with fresh PBS.
Has anyone experienced this problem with NRVMs, or Primaria/gelatin plates in general? If anyone has any advice it would be much appreciated!
Does anyone have any idea about this? It would be much appreciated! Thanks.
Im not much help but I thought I'd tell you your not alone. I am having the same problem after cross-linking MEFs for ChIP using regular 10cm TC dishes. I am going to try scraping them in media and then washing the pellet several times in PBS........we'll see what happens