Improved islet yield and function by use of a chloride channel blocker during collagenase digestion.
Improved islet yield and function by use of a chloride channel blocker during collagenase digestion.
复制标题
在胶原酶消化过程中使用氯离子通道阻滞剂可提高胰岛产量和功能。
DOI：
10.1097/tp.0b013e31822e6eb4
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发表时间：
2011
期刊：
影响因子：
6.2
通讯作者：
Gotoh M.
中科院分区：
文献类型：
--
作者：
Anazawa T;Sato Y;Saito T;Tsuchiya T;Kenjo A;Kimura T;Haga J;Miyake M;Waguri S;Hazama A;Gotoh M.
背景：在分离过程中保护胰岛是胰岛移植成功的必要条件。据报道，氯离子通道抑制可防止各种刺激诱导的细胞死亡。我们研究了氯通道阻断剂4，4′-二异硫氰酸二苯乙烯-2，2 ′-二磺酸二钠盐（DIDS）和细胞外无Cl−条件对胰岛分离结果的影响。和无Cl−基团，葡萄糖酸钠取代氯化钠。我们确定胶原酶消化是否通过Cl−流入细胞诱导胰岛细胞死亡。然后，我们评估胰岛产量和分离isletts.Results的活力。我们观察到胶原酶消化条件下，使用Cl−敏感的荧光染料和随后的胰岛细胞破裂的细胞内Cl−浓度增加。因此，DIDS组和无Cl−组的胰岛产量显著高于对照组，并且前两组的胰岛形态得以保留。在链脲佐菌素诱导的糖尿病严重联合免疫缺陷小鼠中，移植了边缘剂量的胰岛，DIDS组的所有7只小鼠和无Cl−组的7只小鼠中的6只变得血糖正常，而对照组的7只小鼠中只有2只（对照组与DIDS组，P= 0.010;对照组与无Cl−组，P= 0.051）。结论：我们的结果表明，DIDS抑制Cl−流入胰岛在消化过程中保护胰岛，为改善胰岛分离结果提供了新的策略。
Background.Protection of pancreatic islets during isolation procedures is mandatory for successful islet transplantation. Chloride channel inhibition has been reported to prevent cell death induced by various stimuli. We examined the effects of the chloride channel blocker, 4, 4′-diisothiocyanatostilbene-2, 2′-disulfonic acid disodium salt (DIDS) and extracellular Cl−-free conditions on islet isolation outcomes.Methods.Experimental groups were created based on the collagenase solutions used for Wistar rat islet isolation: control group, Hanks' balanced salt solution; DIDS group, 200 μM DIDS; and Cl−-free group, sodium gluconate substituted for sodium chloride. We determined whether collagenase digestion induced the death of islet cells through Cl− influx into the cells. We then assessed islet yield and the viability of isolated islets.Results.We observed an increase in intracellular Cl− concentration under collagenase digestion conditions using a Cl−-sensitive fluorescent dye and subsequent rupture of islet cells. Consequently, islet yields were significantly higher in the DIDS and Cl−-free groups than in the control group, and islet morphology of the former groups was preserved. Of streptozotocin-induced diabetic severe combined immunodeficiency mice transplanted with a marginal dose of islets, all seven mice in the DIDS group and six of the seven mice in the Cl−-free group became normoglycemic, compared with two of seven mice in the control group (control vs. DIDS, P= 0.010; control vs. Cl− free, P= 0.051).Conclusion.Our results indicate that DIDS inhibition of Cl− influx into islets protects islets during digestion procedures, offering a new strategy for the improvement of islet isolation outcomes.