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Long-term culture of human pancreatic slices as a model to study real-time islet regeneration.

Nature communications (2020-07-01)
Mirza Muhammad Fahd Qadir, Silvia Álvarez-Cubela, Jonathan Weitz, Julia K Panzer, Dagmar Klein, Yaisa Moreno-Hernández, Sirlene Cechin, Alejandro Tamayo, Joana Almaça, Helmut Hiller, Maria Beery, Irina Kusmartseva, Mark Atkinson, Stephan Speier, Camillo Ricordi, Alberto Pugliese, Alejandro Caicedo, Christopher A Fraker, Ricardo Luis Pastori, Juan Domínguez-Bendala
ABSTRACT

The culture of live pancreatic tissue slices is a powerful tool for the interrogation of physiology and pathology in an in vitro setting that retains near-intact cytoarchitecture. However, current culture conditions for human pancreatic slices (HPSs) have only been tested for short-term applications, which are not permissive for the long-term, longitudinal study of pancreatic endocrine regeneration. Using a culture system designed to mimic the physiological oxygenation of the pancreas, we demonstrate high viability and preserved endocrine and exocrine function in HPS for at least 10 days after sectioning. This extended lifespan allowed us to dynamically lineage trace and quantify the formation of insulin-producing cells in HPS from both non-diabetic and type 2 diabetic donors. This technology is expected to be of great impact for the conduct of real-time regeneration/developmental studies in the human pancreas.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Dulbecco′s Phosphate Buffered Saline, Modified, without calcium chloride and magnesium chloride, liquid, sterile-filtered, suitable for cell culture
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Phosphate buffered saline, powder, pH 7.4, for preparing 1 L solutions
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Antibiotic Antimycotic Solution (100×), Stabilized, with 10,000 units penicillin, 10 mg streptomycin and 25 μg amphotericin B per mL, 0.1 μm filtered, BioReagent, suitable for cell culture
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Fibronectin bovine plasma, solution, sterile-filtered, BioReagent, suitable for cell culture
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D-(+)-Glucose solution, 100 g/L in H2O, sterile-filtered, BioXtra, suitable for cell culture
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Trypsin inhibitor from Glycine max (soybean), powder, BioReagent, suitable for cell culture
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Aprotinin from bovine lung, BioUltra, 3-8 TIU/mg solid, ≥98% (SDS-PAGE)
Roche
Chymostatin
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Cholecystokinin (CCK) Fragment 26-33 Amide, Non-sulfated, ≥95% (HPLC), powder