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A High-content In Vitro Pancreatic Islet β-cell Replication Discovery Platform
Published on: July 16, 2016
The effect of TRAIL molecule on cell viability in in vitro beta cell culture
I Tekmen1, D Ozyurt, C Pekçetin
1Department of Histology and Embryology, Dokuz Eylul University Medical Faculty, 35340, Inciralti, Izmir, Turkey. isil.tekmen@deu.edu.tr
Abstract:
Insulin-dependent diabetes mellitus (IDDM) is an organ-specific autoimmune disorder triggered by autoreactive T cells directed to pancreas beta-cell antigens. In this disorder, more than 90% of beta cells are destroyed. Cell death may be mediated via soluble or membrane-bound cell death ligands. One of these ligands may be tumour necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL), a member of the TNF-alpha superfamily. In the present study, we examined whether TRAIL had cytotoxic effects on adult rat pancreas beta cell cultures and INS1-E rat insulinoma cell line cultures or not. In this study, cell destruction models were built with TRAIL concentrations of 10, 100 and 1000 ng. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) test was used for evaluating cell viability. It was detected that cell cultures with TRAIL added showed no differences statistically when compared with control cultures containing no toxic additions. These results showed that TRAIL did not have significant cytotoxic effects on pancreas beta cell culture and INS-1E rat insulinoma cell line cultures. Detection of the expression of TRAIL receptors and natural apoptosis inhibitor proteins will be favourable to investigate the resistance mechanisms to TRAIL-induced cell death in this cell culture system.
Insights
Tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) did not induce significant cell death in rat pancreas beta cells or insulinoma cell lines. Further research into resistance mechanisms is recommended.
Area of Science:
- Immunology
- Endocrinology
- Cell Biology
Background:
- Insulin-dependent diabetes mellitus (IDDM) involves autoimmune destruction of pancreatic beta cells.
- Cell death in IDDM may be mediated by ligands like TNF-related apoptosis-inducing ligand (TRAIL).
Purpose of the Study:
- To investigate the cytotoxic effects of TRAIL on adult rat pancreas beta cell cultures and INS1-E rat insulinoma cell line cultures.
Main Methods:
- Cell cultures were exposed to varying concentrations of TRAIL (10, 100, 1000 ng).
- Cell viability was assessed using the 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay.
Main Results:
- TRAIL treatment did not result in statistically significant differences in cell viability compared to control groups.
- TRAIL demonstrated no significant cytotoxic effects on the tested beta cell and insulinoma cell cultures.
Conclusions:
- TRAIL is unlikely to be a primary mediator of beta cell destruction in this experimental system.
- Investigating TRAIL receptor expression and apoptosis inhibitors could elucidate resistance mechanisms to TRAIL-induced cell death.
