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Forschungsdatenbank PMU-SQQUID

Prevention of oxidative stress in porcine islet isolation.
Stiegler, P; Stadlbauer, V; Hackl, F; Schaffellner, S; Iberer, F; Greilberger, J; Strunk, D; Zelzer, S; Lackner, C; Tscheliessnigg, K
J ARTIF ORGANS. 2010; 13(1): 38-47.
Originalarbeiten (Zeitschrift)

PMU-Autor/inn/en

Strunk Dirk

Abstract

High yields of pure and viable porcine islet cells (PICs) to be used for microencapsulation are crucial for successful xenotransplantation. Mechanical disruption of the pancreas, enzymes used for digestion, digestion temperature and time are among the factors known to cause oxidative stress and to impact on the yield, purity and viability of PICs. The aim of our study was to optimize conventional procedures in order to minimize the oxidative stress that occurs during the isolation and purification of PICs. Porcine pancreatic tissue was harvested at a local slaughterhouse, and 15 consecutive isolations of PICs were performed with a modified automated Ricordi method (Graz method) using a shorter digestion time, a lower digestion temperature and minimal mechanical stress. PICs were purified with the Lymphoprep density gradient medium. Purity and viability were assessed immediately after the isolation process and after overnight culture. PIC function was tested in glucose stimulation experiments and insulin concentration was determined by ELISA. Oxidative stress was assessed by measuring isoprostanes (IP), malondialdehyde (MDA) and lipase levels using a HPLC-based, colorimetric liquid assay or ELISA, respectively. The mean yield of PICs was 3479 +/- A 542 IEQs/g pancreas, with 96.4% viability and 97.7% purity. There was no significant loss in PIC viability after overnight culture. Insulin secretion in response to glucose was not impaired after isolation and purification. IP, MDA and lipase levels did not change significantly during the isolation procedure. With our new Graz method we seem to have succeeded in preventing oxidative stress and achieving high yields of pure and viable PICs.


Useful keywords (using NLM MeSH Indexing)

Analysis of Variance

Animals

Cell Culture Techniques/methods*

Enzyme-Linked Immunosorbent Assay

Insulin/secretion

Islets of Langerhans/metabolism

Islets of Langerhans/pathology

Islets of Langerhans/secretion

Islets of Langerhans Transplantation/methods*

Islets of Langerhans Transplantation/pathology

Oxidative Stress*

Swine

Transplantation, Heterologous/methods*

Transplantation, Heterologous/pathology


Find related publications in this database (Keywords)

Islet transplantation
Islet isolation
Xenotransplantation
Oxidative stress
Isoprostanes