Brown J, Kemp J A, Hurt S, Clark W R
Diabetes. 1980;29 Suppl 1:70-3. doi: 10.2337/diab.29.1.s70.
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.
为确定成功冷冻保存人胎儿胰腺的最佳条件，对大鼠器官所采用的技术进行了改进。研究的参数包括接触冷冻保护剂二甲基亚砜（DMSO）的条件、冷却和解冻速率以及体外培养的影响。总共研究了33个因前列腺素诱导流产导致胎儿死亡后获取的胰腺。胰腺的存活基于在4小时孵育期间，胰腺组织块（2立方毫米）将3H-氨基酸掺入蛋白质的情况，并与来自同一胰腺的未冷冻对照组织块进行比较。发现在37摄氏度下暴露4小时后，DMSO的毒性很强。改变温度、暴露时间和DMSO浓度对冻融后存活的影响表明，室温下1.5M DMSO处理1小时是最佳的。冷却速率为0.22摄氏度/分钟，在室温下解冻。由于这些条件仅导致50%的存活率，因此在接触DMSO之前增加了一段培养时间。最佳培养持续时间为12 - 16小时。采用此方法并在解冻后添加24小时培养，存活率在对照的70%至120%之间变化。如果冻融后的胰腺生长和胰岛素产生功能测试呈阳性，人胎儿胰腺的永久性短缺将成为可能。
