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体外及异种移植中琼脂糖聚苯乙烯磺酸微囊化猪胰岛的功能评估

Functional assessment of microencapsulated porcine islets with agarose polystyrene sulfonic acid in vitro and in xenotransplantation.

作者信息

Tashiro H, Iwata H, Warnock G L, Tanigawa M, Ototani Y, Tsuji T

机构信息

Department of Surgery, The British Columbia University, Canada.

出版信息

Transplant Proc. 2005 Oct;37(8):3512-3. doi: 10.1016/j.transproceed.2005.09.133.

Abstract

We evaluated the functional efficacy of microencapsulated porcine islet xenografts transplanted into nonobese diabetic (NOD) mice. Islets were isolated from the pancreata of CSK miniature swine by manual collagenase digestion and Ficoll purification. Purified porcine islets were immediately encapsulated into microbeads of agarose polystyrene sulfonic acid (Ag-PSSa). They remained morphologically intact by dithizone staining after 7 days in culture. Insulin secretion from encapsulated islets was determined in response to glucose challenge during perifusion. When encapsulated islets were exposed to 200 mg/dL glucose, within 5 minutes, insulin release became 5-fold greater than that at 80 mg/dL. However, a second phase insulin secretion appeared in response to 250 mg/dL glucose challenge. In xenotransplantation, microencapsulated porcine islets (1000 to 1800 MC islets) were transplanted into the peritoneal cavity of diabetic NOD mice (n = 4) without immunosuppression. The survival times after the onset of diabetes were observed after both MC islets transplanted NOD mice and nontransplanted NOD mice (n = 4). MC islets transplant recipients had significantly (P < .05) longer survival (47.5 +/- 18.6; mean +/- SD) than nontransplanted NOD mice (21.0 +/- 9.31), although random blood glucose levels were not normalized.

摘要

我们评估了移植到非肥胖糖尿病(NOD)小鼠体内的微囊化猪胰岛异种移植物的功能疗效。通过手工胶原酶消化和Ficoll纯化从CSK小型猪的胰腺中分离胰岛。将纯化的猪胰岛立即封装到琼脂糖聚苯乙烯磺酸(Ag-PSSa)微珠中。培养7天后,经双硫腙染色,它们的形态保持完整。在灌注过程中,测定了微囊化胰岛对葡萄糖刺激的胰岛素分泌。当微囊化胰岛暴露于200mg/dL葡萄糖时,5分钟内胰岛素释放量比80mg/dL时增加了5倍。然而,在250mg/dL葡萄糖刺激下出现了第二阶段胰岛素分泌。在异种移植中,将微囊化猪胰岛(1000至1800个微囊化胰岛)在不进行免疫抑制的情况下移植到糖尿病NOD小鼠(n = 4)的腹腔中。观察了微囊化胰岛移植的NOD小鼠和未移植的NOD小鼠(n = 4)糖尿病发病后的存活时间。微囊化胰岛移植受体的存活时间(47.5 +/- 18.6;平均值 +/- 标准差)明显(P <.05)长于未移植的NOD小鼠(21.0 +/- 9.31),尽管随机血糖水平未恢复正常。

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