Monolayer cultures were prepared by dissociating pancreases from newborn rats with a trypsin—collagenase solution. Cultures consisted of numerous discrete colonies containing beta cells, with a network of fibroblastoid cells between colonies. Large amounts of immunoreactive insulin (60–70 mU per culture per 2 days) were released into the culture medium. Active beta cell replication was documented by radioautography using thymidine–3H, and by incubation with Colcemid. Both the release of insulin and the frequency of beta cell replication were greater at a glucose concentration of 300 mg/100 ml medium compared to 100 mg/100 ml medium. Even after two months, cultures maintained at 300 mg/100 ml glucose continued to release large amounts of insulin into the medium indicating that beta cells remain viable in monolayer cultures for extended periods of time, and do not undergo exhaustion atrophy in the presence of a high glucose concentration. (Endocrinology92: 212, 1973)

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