INSULIN RELEASE FROM HUMAN FOETAL PANCREAS IN TISSUE CULTURE

Abstract

SUMMARY Various methods of tissue culture were studied in an attempt to grow human foetal pancreas under conditions favourable for insulin release. Simple dicing of pancreas was superior to plasma clot adhesion or collagenase digestion for the preparation of tissue for culture. The culture medium described by Kahri (1966) (containing 25% heated postnatal calf serum) was the most suitable of four tested for the study of insulin release from the tissue culture. In this medium insulin could be measured quantitatively by radioimmunoassay and insulin degradation occurred at the rate of 20–25%/24 h. In other media the pancreas grew less well or insulin degradation was much greater. Human foetal pancreas grown under optimal conditions released insulin for up to 34 days. Insulin released into the culture medium did not appear to inhibit the further release of insulin. In some experiments the total amount of insulin released into the culture medium was several-fold greater than that in the pancreas originally seeded. In acute incubation experiments barium and theophylline stimulated insulin release from pancreas cultures. It proved impossible to identify the cells from which insulin was released but they did not appear to be in the monolayer which was composed of fibroblasts.