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Abstract:
To determine the optimal conditions for successful cryopreservation of the human fetal pancreas, techniques developed for the rat organ were modified. The parameters studied were the conditions of exposure to the cryopreservation agent dimethyl sulfoxide (dmso), the rate of cooling and thawing, and the effect of in vitro culture. A total of 33 pancreases, obtained after fetal death due to prostaglandin-induced abortion, was studied. Survival of the pancreas is based on incorporation of 3H-amino acids into protein by pancreas pieces (2 mm3) during a 4-h incubation compared with nonfrozen control pieces from the same pancreas. Toxicity of DMSO at 37 degrees C was found to be severe after a 4-h exposure. Varying the effects of temperature, time of exposure, and concentrations of DMSO on survival after freeze-thaw revealed that 1.5 M DMSO for 1 h at room temperature was optimal. The cooling rate was 0.22 degrees C/min and thawing was at room temperature. Since these conditions resulted in only 50% survival, a period of culture before exposure to DMSO was added. The optimal duration of culture was 12--16 h. Using this method with addition of culture for 24 h after thawing, survival has varied from 70 to 120% of control. If a functional test for growth and insulin production by the frozen-thawed pancreas is positive, permanent shortage of the human fetal pancreas will be possible.