Improved Differentiation Towards Insulin Producing Beta-Cells Derived from Healthy Canine Pancreatic Ductal Organoids.

BACKGROUND

Diabetes mellitus (DM) is a common potentially life-threatening endocrine disorder in pets and humans. Since only symptomatic treatment is available, a more sustainable treatment is urgently needed.

OBJECTIVE

The aim of this study is to establish functional differentiated canine pancreatic β-cells that release insulin upon glucose stimulus.

METHODS

Pancreatic tissue was obtained from surplus material of healthy dogs (n = 4), euthanized for non-pancreatic related research. Ductal cells were isolated and expanded in dog pancreas expansion media (dpEM) and differentiated and maturated in five sequentially added pancreas differentiation media (PDMs). Gene expression was analyzed by reversed transcriptase qPCR (RT-qPCR), and insulin release was analyzed with a canine-specific ELISA.

RESULTS

Canine pancreatic ductal cells ( and expression) were differentiated into β-cells expressing key β-cell-related genes: , and low levels of . Neither (α-cells) nor and were expressed, and was expressed at low levels. The differentiated cells released insulin upon glucose stimulation.

CONCLUSION AND IMPLICATIONS

The step-by-step differentiation protocol, mimicking pancreatic organogenesis, resulted in β-cells secreting insulin levels suitable for β-cell disease modelling. It remains to be seen if stem cells from diseased animals behave similarly.