Primary gastric epithelial cells - (May/23/2008 )
I intend to isolate primary gastric epithelial cells from mouse stomach. (These cells will be used for infection with H.pylori). But I do not have an experience in this area. Could tell me please about the main points in this procedure that must be taken into consideration.
Also I would like to ask you, is it important to coat the plastic with collagen, or Matrigel. I have read about the Extracel Hydrogels. Does it work well?Could it serve as alternative to Matrigel?
-Roman80-
QUOTE (Roman80 @ May 23 2008, 02:34 AM)
I intend to isolate primary gastric epithelial cells from mouse stomach. (These cells will be used for infection with H.pylori). But I do not have an experience in this area. Could tell me please about the main points in this procedure that must be taken into consideration.
Also I would like to ask you, is it important to coat the plastic with collagen, or Matrigel. I have read about the Extracel Hydrogels. Does it work well?Could it serve as alternative to Matrigel?
Also I would like to ask you, is it important to coat the plastic with collagen, or Matrigel. I have read about the Extracel Hydrogels. Does it work well?Could it serve as alternative to Matrigel?
it is a good idea to infect native cells instead of oncogenic cell lines;
there are some protocols to isolate exocrine gastric cells, most famous is Lewin-protocol; you cut off the stomach, invert it, bind it at both ends, and inject a buffer with proteases such as collagenase and/or pronase; inverted stomachs are incubated in warmed buffer under stirring; you collect the cells and start culturing; Matrigel is often used
-The Bearer-