Intestinal Cell Isolation Protocol?

Stephen H Cheng scheng at rice.edu
Sun Nov 20 03:56:37 EST 1994


	Hi, I'm trying to isolate hamster ileum cells for some binding 
assays, and I was wondering if anybody here has some experience with 
getting these guys out alive?

	I'm currently using a method from:

"Intestinal Epithelial Cell Surface Membrane Glycoprotein Synthesis."  
Weiser, Milton.  The Journal of Biological Chemistry.  Vol. 248 (7).
pp-2536-2541.

which mainly involves the treating of the intestine with a solution of:
	1.5 mM KCl
	96 mM NaCl
	27 mM Sodium Citrate
	  8 mMKH2PO4
	 5.6 mM Na2HPO4
	pH of 7.3

and then incubation with:
PBS (w/o Ca2+ or Mg2+)
1.5 mM EDTA
.5 mM dithiothreitol

	Although my yield of cells is nice, they consistently fail my 
trypan blue exclusion test for viability, and I would kinda like them alive.

Any advice or help through this post or e-mail (micsnc at ttuhsc.edu) would 
be appreciated.

--SC



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