Isolation and cryopreservation of human pancreatic islets

The outlined methods were able to isolate relatively large number of intact pancreatic islets from human cadaver pancreata. Confirmation of the nature of the endocrine and exocrine tissue was provided by electron microscopy. The islets were able to tolerate a 14 day period of storage in liquid N2 when frozen and thawed by the protocol given. Viability of the islets both before and after the freeze-thaw sequence was established by secretion of insulin in response to stimulation with both theophylline and high glucose in a dynamic perifusion system. The ability of the islets to survive a 24 hr. period of tissue culture after the freeze/thaw sequence and again respond with insulin secretion in response to hyperglycemia or theophylline stimulation confirms their viability and metabolic activity.