Islet isolation assessment in man and large animals

Camillo Ricordi, Derek W R Gray, Bernhard J. Hering, Dixon B. Kaufman, Garth L. Warnock, Norman M. Kneteman, Stephen P. Lake, Nicholas J M London, Carlo Socci, Rodolfo Alejandro, Yijun Zeng, David W. Scharp, Giorgio Viviani, Luca Falqui, Andreas Tzakis, Reinhard G. Bretzel, Konrad Federlin, Guido Pozza, Roger F L James, Ray V. RajotteValerio Di Carlo, Peter J. Morris, David E R Sutherland, Thomas E. Starzl, Daniel H. Mintz, Paul E. Lacy

Research output: Contribution to journalArticle

463 Citations (Scopus)

Abstract

Recent progress in islet isolation from the pancreas of large mammals including man, accentuated the need for the development of precise and reproducible techniques to assess islet yield. In this report both quantitative and qualitative criteria for islet isolation assessment were discussed, the main topics being the determination of number, volume, purity, morphologic integrity and in vitro and in vivo function tests of the final islet preparations. It has been recommended that dithizone should be used as a specific stain for immediate detection of islet tissue making it possible to estimate both the total number of islets (dividing them into classes of 50 μ diameter range increments) and the purity of the final preparation. Appropriate morphological assessment should include confirmation of islet identification, assessment of the morphological integrity and of the purity of the islet preparation. The use of fluorometric inclusion and exclusion dyes together have been suggested as a viability assay to simultaneously quantitate the proportion of cells that are intact or damaged. Perifusion of islets with glucose provides a dynamic profile of glucose-mediated insulin release and of the ability of the cells to down regulate insulin secretion after the glycemic challenge is interrupted. Although perifusion data provides a useful guide to islet viability the quantity and kinetics of insulin release do not necessarily predict islet performance after implantation. Therefore, the ultimate test of islet viability is their function after transplantation into a diabetic recipient. For this reason, in vivo models of transplantation of an aliquot of the final islet preparation into diabetic nude (athymic) rodents have been suggested. We hope that these general guidelines will be of assistance to standardize the assessment of islet isolations, making it possible to better interpret and compare procedures from different centers.

Original languageEnglish
Pages (from-to)185-195
Number of pages11
JournalActa Diabetologica Latina
Volume27
Issue number3
DOIs
StatePublished - Jul 1 1990

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Insulin
Coloring Agents
Transplantation
Dithizone
Glucose
Pancreas
Mammals
Rodentia
Down-Regulation
Guidelines
In Vitro Techniques

Keywords

  • Assessment
  • Islet
  • Isolation

ASJC Scopus subject areas

  • Internal Medicine
  • Endocrinology, Diabetes and Metabolism
  • Endocrinology

Cite this

Ricordi, C., Gray, D. W. R., Hering, B. J., Kaufman, D. B., Warnock, G. L., Kneteman, N. M., ... Lacy, P. E. (1990). Islet isolation assessment in man and large animals. Acta Diabetologica Latina, 27(3), 185-195. https://doi.org/10.1007/BF02581331

Islet isolation assessment in man and large animals. / Ricordi, Camillo; Gray, Derek W R; Hering, Bernhard J.; Kaufman, Dixon B.; Warnock, Garth L.; Kneteman, Norman M.; Lake, Stephen P.; London, Nicholas J M; Socci, Carlo; Alejandro, Rodolfo; Zeng, Yijun; Scharp, David W.; Viviani, Giorgio; Falqui, Luca; Tzakis, Andreas; Bretzel, Reinhard G.; Federlin, Konrad; Pozza, Guido; James, Roger F L; Rajotte, Ray V.; Carlo, Valerio Di; Morris, Peter J.; Sutherland, David E R; Starzl, Thomas E.; Mintz, Daniel H.; Lacy, Paul E.

In: Acta Diabetologica Latina, Vol. 27, No. 3, 01.07.1990, p. 185-195.

Research output: Contribution to journalArticle

Ricordi, C, Gray, DWR, Hering, BJ, Kaufman, DB, Warnock, GL, Kneteman, NM, Lake, SP, London, NJM, Socci, C, Alejandro, R, Zeng, Y, Scharp, DW, Viviani, G, Falqui, L, Tzakis, A, Bretzel, RG, Federlin, K, Pozza, G, James, RFL, Rajotte, RV, Carlo, VD, Morris, PJ, Sutherland, DER, Starzl, TE, Mintz, DH & Lacy, PE 1990, 'Islet isolation assessment in man and large animals', Acta Diabetologica Latina, vol. 27, no. 3, pp. 185-195. https://doi.org/10.1007/BF02581331
Ricordi C, Gray DWR, Hering BJ, Kaufman DB, Warnock GL, Kneteman NM et al. Islet isolation assessment in man and large animals. Acta Diabetologica Latina. 1990 Jul 1;27(3):185-195. https://doi.org/10.1007/BF02581331
Ricordi, Camillo ; Gray, Derek W R ; Hering, Bernhard J. ; Kaufman, Dixon B. ; Warnock, Garth L. ; Kneteman, Norman M. ; Lake, Stephen P. ; London, Nicholas J M ; Socci, Carlo ; Alejandro, Rodolfo ; Zeng, Yijun ; Scharp, David W. ; Viviani, Giorgio ; Falqui, Luca ; Tzakis, Andreas ; Bretzel, Reinhard G. ; Federlin, Konrad ; Pozza, Guido ; James, Roger F L ; Rajotte, Ray V. ; Carlo, Valerio Di ; Morris, Peter J. ; Sutherland, David E R ; Starzl, Thomas E. ; Mintz, Daniel H. ; Lacy, Paul E. / Islet isolation assessment in man and large animals. In: Acta Diabetologica Latina. 1990 ; Vol. 27, No. 3. pp. 185-195.
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