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

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
摘要

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.

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Sigma-Aldrich
杜氏磷酸盐缓冲盐水, 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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抗生素抗霉菌溶液(100×),稳定, 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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纤连蛋白牛血浆, solution, sterile-filtered, BioReagent, suitable for cell culture
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胰蛋白酶抑制剂 来源于大豆, powder, BioReagent, suitable for cell culture
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抑肽酶 来源于牛肺, BioUltra, 3-8 TIU/mg solid, ≥98% (SDS-PAGE)
Roche
胰凝乳蛋白酶抑制剂
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Cholecystokinin (CCK) Fragment 26-33 Amide, Non-sulfated, ≥95% (HPLC), powder