Human Pancreatic Islet-Cell Culture: Different Responses in Insulin Release for Fetal and Adult Pancreatic Cells

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Both fetal and adult human pancreatic tissues were digested with collagenase, and the resulting cell clumps, including isolated islets, were cultured with Dulbecco's modified Eagle medium (D-MEM) containing 10 fetal bovine serum (FBS) in a CO2 incubator gassed with 5 % CO2 in air. Cells became attached to the bottoms of the petri dishes and formed cell sheets after 3 days. Most epithelial cells in the monolayer were positively stained with aldehyde fuchsin and were maintained for about 2 weeks.<BR>Insulin release in the fetal culture on the 3rd day was 1.3 times for 3 mg/ml glucose, 3.5 times for 150 μg/m1 tolbutamide and 7.8 times for 10 mM theo-phylline, as compared to that of 1 mg/ml glucose. However, in the adult cul-ture, the stimulus of glucose, tolbutamide and theophylline resulted in equal increases in insulin release of about 5 to 6 times. These results indicate that the cell function, characteristic of fetal and adult human islets in vivo, is preserved in culture.<BR>In the fetal culture, the insulin response to glucose tended to increase on the 14th day, whereas the response to theophylline and tolbutamide decreased, evidence of the possible development of a B cell function in the culture.

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