Abstract
There is an unmet medical need for the improvement of pancreatic islet maintenance in culture. Due to restricted donor availability it is essential to ameliorate islet viability and graft engraftment. The aim of this study was to compare the standard tissue culture techniques with the advanced Real Architecture For 3D Tissue (RAFT™) culture system in terms of viability and hormone production. Here, we first report that islets embedded in RAFT™ collagen type I advanced tissue culture system maintain their tissue integrity better than in monolayer and suspension cultures. The Calcein violet assay and Annexin V/propidium-iodide staining show higher cell viability in the RAFT™ culture system. Quantitative real-time PCR data showed that RAFT™ increases insulin expression after 18 days in culture compared to traditional methods. Enhanced insulin and glucagon production was further verified by immunofluorescent staining in a time-course manner. These results indicate that RAFT™ tissue culture platform can be a promising tool to maintain pancreatic islet spheroid integrity and culture islets for downstream high throughput pharmacological studies ex vivo.
Highlights
Enhanced insulin and glucagon production was further verified by immunofluorescent staining in a time-course manner. These results indicate that RAFTTM tissue culture platform can be a promising tool to maintain pancreatic islet spheroid integrity and culture islets for downstream high throughput pharmacological studies ex vivo
Gestational diabetes occurs in approximately 16% of pregnant woman and tend to progress to type 2 Diabetes mellitus (DM) for 15–50% of the patients within 5 years but in most of the cases can elapse by physical activity and dietary change (Gilinsky et al 2015)
In line with the literature, mouse pancreatic islets consisted of more insulin producing b-cells (Fig. 2c) than glucagon producing a-cells (Fig. 2a) (Dolensek et al 2015)
Summary
Almost 90–95% of adult patients suffer from type 2 DM (Lysy et al 2016). Both genetic and life style elements (obesity, unhealthy diet, low physical activity, stress and smoking) can contribute to the establishment of type 2 DM where muscle and adipose tissues are resistant to insulin produced by intact b-cells (Tsasis et al 2016). Kidney clinical transplantation results in 81.5 or 89% 1 year survival, respectively, the 10 year survival currently is around 75% only, further research is much needed to improve b-cell replacement in insulin dependent diabetes (Lysy et al 2016) Pancreas or pancreas ? kidney clinical transplantation results in 81.5 or 89% 1 year survival, respectively, the 10 year survival currently is around 75% only, further research is much needed to improve b-cell replacement in insulin dependent diabetes (Lysy et al 2016)
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