i try to isolate mouse crypt with this method:1,cut off duodenum and discard pancreas;2, rinse the intestine with cold PBS containing 50μg/ml gentamicin about 10 times ; 3, cut this segment into smaller portions and digest with 2mM PBS-EDTA for 30minutes, 2 times ;4, extract the supernate and pass by 70μM filter; 5, centrifuge the collected suspension with 200xg,4℃,10 min; 6, resuspend the pallet with DMEM/F12+10%FBS+ 0.1%Penicillin - Streptomyces, cultrue under at 37℃，5%CO2