The pellets were washed with DMEM c

The pellets were washed with DMEM containing 2% FBS and 2%
sorbitol, and the supernatant containing the purified crypt fraction
was collected by centrifugation at 2506 g at room temperature
(RT). The crypts were then resuspended and incubated in
complete DMEM. In each case, the medium with the non-
adhering cells was recovered after 90-min culture and plated into a
new plastic culture flask. Medium was changed every 48 h and the
confluence reached within approximately 8 days. Subcultures were
performed after trypsinization (0.5% trypsin-0.54 mM EDTA in
PBS, at 23uC for 5 min).