cbil.upenn.edu:RAD.Protocol:4814:Protocol - cbil.upenn.edu:RAD.Protocol:4814:Protocol
For best results use Trypsin that has only been freeze/thawed once. Remove pancreas and place in ice cold PBS. Separate GFP+ pancreas from GFP- pancreas. Mince the pancreas in scintillation vials of trypsin (pre-warm 37 degrees C). Use fine scissors and finely chop pancreas. Add stir bar and incubate in 37 degrees C water bath for 7-15 min with the stir plate set to 1. Terminate the trypsin by adding an equal volume of RPMI + 10% FBS and place immediately on ice. Pass cells through cell strainer. Gently touch the membrane to allow the flow and pipette up and down. If filter clogs, use a new one. GFP+ cells were sorted at the Penn Flow Cytometry and Cell Sorting Resource Laboratory.