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Protocol for Studying Reprogramming of Mouse Pancreatic Acinar Cells to β-like Cells.

STAR protocols (2020-10-29)
Ofer Elhanani, Michael D Walker
ABSTRACT

The potential of reprogrammed β cells derived from pancreatic exocrine cells to treat diabetes has been demonstrated in animal models. However, the precise mechanisms and regulators involved in this process are not clear. Here, we describe a method that allows mechanistic studies of this process in primary exocrine cultures using adenoviral expression vectors. This rapid 5-day protocol, provides the researcher with a highly controlled experimental system in which the effects of different compounds or genetic manipulations can be studied. For complete details on the use and execution of this protocol, please refer to Elhanani et al. (2020).

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Collagen, Type I solution from rat tail, BioReagent, suitable for cell culture, sterile-filtered
Sigma-Aldrich
Hanks′ Balanced Salt solution, Modified, with sodium bicarbonate, without phenol red, calcium chloride and magnesium sulfate, liquid, sterile-filtered, suitable for cell culture
Sigma-Aldrich
SIGMAFAST 3,3′-Diaminobenzidine tablets, tablet, To prepare 5 mL
Sigma-Aldrich
4′,6-Diamidino-2-phenylindole dihydrochloride, BioReagent, suitable for fluorescence, ≥95.0% (HPLC)
Sigma-Aldrich
Bovine Serum Albumin, heat shock fraction, suitable for RIA, pH 5.2, ≥96%
Sigma-Aldrich
Ethylene glycol-bis(2-aminoethylether)-N,N,N′,N′-tetraacetic acid, ≥97.0%
Sigma-Aldrich
Trypsin inhibitor from Glycine max (soybean), lyophilized powder