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
