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未纯化胰岛组织的体外和体内活力评估

In vitro and in vivo viability assessment of unpurified pancreatic islet tissue.

作者信息

Hesse U J, Schmitz-Rode M, Danis J, Tunggal B, Meyer G, Weyer J, Pichlmaier H

机构信息

Department of Surgery, University of Cologne, Germany.

出版信息

J Surg Res. 1994 Nov;57(5):556-62. doi: 10.1006/jsre.1994.1182.

Abstract

The viability of porcine collagenase-prepared islet preparations (n = 16) was classified by 31P-NMR spectroscopy, staining by neutral red and trypan blue, and in vitro insulin secretion following glucose challenge. Vital islets exhibited a phosphate diester/phosphate monoester (PDE/PME) ratio of 0.5-0.9, a staining score of 18-30 and an insulin secretion responding well to glucose challenge. Damaged islets performed at a PDE/PME of 0.2-0.49 and a staining score of 9-17 and necrotic islets had 0.0-0.49 and a staining score of 9-17 and necrotic islets had 0.0-0.19 and 0-8, respectively. The islets of the latter two groups did not adequately respond to glucose. The in vivo function following autotransplantation of these islets into the spleen was investigated in five recipients of more than 3000/kg vital islets of which 4 expressed daily normoglycemia (< 200 mg%), normalized intravenous glucose tolerance (K = -2.21), and a prolonged survival (mean +/- SD) of 167 +/- 12 days compared to five recipients of > 3000/kg damaged islets (K = -0.814) (P = 0.0017) and a survival of 86 +/- 21 days (P = 0.0096). It is suggested that 31P-NMR spectroscopy is a valuable and practical method to predict islet graft viability prior to transplantation in order to assure good graft function in the recipient.

摘要

采用31P-NMR光谱法、中性红染色法和台盼蓝染色法,以及葡萄糖刺激后的体外胰岛素分泌情况,对猪胶原酶制备的胰岛制剂(n = 16)的活力进行分类。有活力的胰岛磷酸二酯/磷酸单酯(PDE/PME)比值为0.5 - 0.9,染色评分为18 - 30,且胰岛素分泌对葡萄糖刺激反应良好。受损胰岛的PDE/PME为0.2 - 0.49,染色评分为9 - 17,坏死胰岛的PDE/PME为0.0 - 0.49,染色评分为9 - 17,坏死胰岛的染色评分为0.0 - 0.19和0 - 8。后两组胰岛对葡萄糖的反应不佳。将这些胰岛自体移植到脾脏后,对5名接受超过3000/kg有活力胰岛移植的受体的体内功能进行了研究,其中4名受体每日血糖正常(< 200 mg%),静脉葡萄糖耐量正常(K = -2.21),平均存活时间(均值±标准差)为167 ± 12天;与之相比,5名接受超过3000/kg受损胰岛移植的受体(K = -0.814)(P = 0.0017),存活时间为86 ± 21天(P = 0.0096)。研究表明,31P-NMR光谱法是一种在移植前预测胰岛移植活力的有价值且实用的方法,以确保受体中移植功能良好。

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