Ishii Show, Saito Takuro, Ise Kazuya, Yamashita Michitoshi, Sato Yoshihiro, Saito Takaharu, Tsukada Manabu, Oshibe Ikuro, Kenjo Akira, Kimura Takashi, Anazawa Takayuki, Suzuki Shigeya, Gotoh Mitsukazu
Department of Surgery; Fukushima Medical University; Fukushima City, Fukushima, Japan.
Research and Development Division; Kikkoman Co., Ltd; Noda City, Chiba, Japan.
Islets. 2012 Jan-Feb;4(1):32-9. doi: 10.4161/isl.18607. Epub 2012 Jan 1.
Culture of islets prior to transplantation needs to be revisited for maintaining functional islet capacity. This study was conducted to compare cold UW (University of Wisconsin) preservation with conventional culture based on insulin secretory capacity in vitro and in vivo. Islets isolated from Wistar rats were either cultured for 24 h at 37°C in RPMI1640 medium or DMEM containing various concentrations of glucose or preserved for the same period in UW solution or in DMEM solution at 4°C. The islet yield in UW group, but not in other groups, was maintained as comparable with that of fresh islets. Insulin secretory capacity in response to glucose was maintained only in the islets of UW group, but not in other groups. SCID mice given 300 IEQ islets of UW group showed gradual restoration of normoglycemia as found in the mice given freshly isolated islets. Meanwhile, those mice given cultured islets for 24 h at 37°C in RPMI1640 medium showed rapid decrease of blood glucose levels on day 1 followed by relatively elevated levels on day 2, suggesting unstable insulin secretory capacity of islets. Morphological staining with anti-HMGB1 (high mobility group B1) antibody revealed central damage of islets in all culture groups regardless of glucose concentration and in islets of cold DMEM group, whereas those in the UW group were quite intact. These results suggest that cold preservation in UW solution is simple and beneficial in protecting islets morphologically and functionally before transplantation.
为维持胰岛的功能容量,移植前胰岛的培养方法需要重新审视。本研究旨在基于体外和体内胰岛素分泌能力,比较冷UW(威斯康星大学)保存法与传统培养法。从Wistar大鼠分离的胰岛,要么在含有不同浓度葡萄糖的RPMI1640培养基或DMEM中于37°C培养24小时,要么在UW溶液或DMEM溶液中于4°C保存相同时间。UW组的胰岛产量得以维持,与新鲜胰岛相当,而其他组则不然。仅UW组的胰岛对葡萄糖的胰岛素分泌能力得以维持,其他组则未维持。给予UW组300个胰岛等效物(IEQ)的SCID小鼠,血糖逐渐恢复正常,如同给予新鲜分离胰岛的小鼠。与此同时,那些在RPMI1640培养基中于37°C培养24小时的培养胰岛的小鼠,第1天血糖水平迅速下降,随后第2天相对升高,提示胰岛胰岛素分泌能力不稳定。用抗HMGB1(高迁移率族蛋白B1)抗体进行形态学染色显示,所有培养组的胰岛中央均有损伤,无论葡萄糖浓度如何,冷DMEM组的胰岛也有损伤,而UW组的胰岛相当完整。这些结果表明,UW溶液冷保存法简单且有利于在移植前从形态和功能上保护胰岛。