Human islet function following 20 years of cryogenic biobanking
Human islet function following 20 years of cryogenic biobanking
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DOI：
10.1007/s00125-015-3598-4
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发表时间：
2015-07-01
期刊：
影响因子：
8.2
通讯作者：
MacDonald, Patrick E.
中科院分区：
文献类型：
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作者：
Fox, Jocelyn E. Manning;Lyon, James;MacDonald, Patrick E.
目的/假设 低温（-196 摄氏度）孤立胰岛储存具有潜在优势，包括维护可行的胰岛以供未来研究。因此，我们评估了冷冻保存近20年的胰岛的体外和体内功能。方法从1991年至2001年冷冻保存人类胰岛，并于2012年至2014年解冻。通过免疫染色、膜片钳电生理学、胰岛素分泌、转录组分析和移植到链脲佐菌素（STZ）诱导的糖尿病小鼠模型中对这些胰岛进行表征。时间为 17.6 +/- 0.4 年 (n = 43)。解冻的胰岛双硫腙染色呈阳性，含有胰岛素阳性和胰高血糖素阳性细胞，并且显示出与新鲜分离的胰岛相似的细胞凋亡水平和转录组谱，尽管其胰岛素含量较低。冷冻保存的β细胞具有与新鲜分离的β细胞相同的离子通道和胞吐反应。来自五个供体的子集的细胞在冷冻保存前后表现出相似的灌注胰岛素分泌特征。将冷冻保存的胰岛移植到糖尿病小鼠体内改善了它们的葡萄糖耐量，但并没有使它们的血糖水平完全正常化。移植后 10 周可检测到循环人胰岛素和胰岛素阳性移植物。结论/解释我们已经证明了长期储存人类胰岛用于研究的潜力，这可以利用未来技术使用大量捐赠者的组织来获得对糖尿病的新见解。
Aims/hypothesis There are potential advantages to the low-temperature (-196 degrees C) banking of isolated islets, including the maintenance of viable islets for future research. We therefore assessed the in vitro and in vivo function of islets cryopreserved for nearly 20 years.Methods Human islets were cryopreserved from 1991 to 2001 and thawed between 2012 and 2014. These were characterised by immunostaining, patch-clamp electrophysiology, insulin secretion, transcriptome analysis and transplantation into a streptozotocin (STZ)-induced mouse model of diabetes.Results The cryopreservation time was 17.6 +/- 0.4 years (n = 43). The thawed islets stained positive with dithizone, contained insulin-positive and glucagon-positive cells, and displayed levels of apoptosis and transcriptome profiles similar to those of freshly isolated islets, although their insulin content was lower. The cryopreserved beta cells possessed ion channels and exocytotic responses identical to those of freshly isolated beta cells. Cells from a subset of five donors demonstrated similar perifusion insulin secretion profiles pre- and post-cryopreservation. The transplantation of cryopreserved islets into the diabetic mice improved their glucose tolerance but did not completely normalise their blood glucose levels. Circulating human insulin and insulin-positive grafts were detectable at 10 weeks post-transplantation.Conclusions/interpretation We have demonstrated the potential for long-term banking of human islets for research, which could enable the use of tissue from a large number of donors with future technologies to gain new insight into diabetes.