Fetal progenitor cells derived from human 10-12-week fetal pancreases.
The pancreatic tissue is digested with collagenase XI. Extracted islet-like structures are suspended in Dulbecco's Modified Eagle Medium:Nutrient Mixture F-12 (DMEM/F12) in a cell culture dish. After slow, manual shaking of the dish, the islet-like structures are moved to the center of the dish and picked using a pipette, under a stereomicroscope. The islet-like structures are resuspended and cultured in a 37°C, 5% CO2 incubator in MEM/F12 medium containing fetal calf serum for stem cells (5%), leukemia inhibitory factor (LIF; 40 μg/L), basic fibroblast growth factor (bFGF; 10 μg/L), epidermal growth factor (EGF; 10 μg/L), penicillin (105 U/L) and streptomycin (100 mg/L). Adherent cells that grow from the islet-like structures after 24 hours are trypsinized and passaged with trypsin/EDTA solution at confluence.