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Long-Term Expansion of Pancreatic Islet Organoids from Resident Procr+ Progenitors.

Cell (2020-03-24)
Daisong Wang, Jingqiang Wang, Lanyue Bai, Hong Pan, Hua Feng, Hans Clevers, Yi Arial Zeng
ZUSAMMENFASSUNG

It has generally proven challenging to produce functional β cells in vitro. Here, we describe a previously unidentified protein C receptor positive (Procr+) cell population in adult mouse pancreas through single-cell RNA sequencing (scRNA-seq). The cells reside in islets, do not express differentiation markers, and feature epithelial-to-mesenchymal transition characteristics. By genetic lineage tracing, Procr+ islet cells undergo clonal expansion and generate all four endocrine cell types during adult homeostasis. Sorted Procr+ cells, representing ∼1% of islet cells, can robustly form islet-like organoids when cultured at clonal density. Exponential expansion can be maintained over long periods by serial passaging, while differentiation can be induced at any time point in culture. β cells dominate in differentiated islet organoids, while α, δ, and PP cells occur at lower frequencies. The organoids are glucose-responsive and insulin-secreting. Upon transplantation in diabetic mice, these organoids reverse disease. These findings demonstrate that the adult mouse pancreatic islet contains a population of Procr+ endocrine progenitors.

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Produktbeschreibung

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HEPES, BioPerformance Certified, ≥99.5% (titration), suitable for cell culture
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Formaldehyd -Lösung, for molecular biology, 36.5-38% in H2O
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Triton X-100, laboratory grade
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TERGITOL -Lösung, Type NP-40, 70% in H2O
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Nocodazol, ≥99% (TLC), powder
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Glutaraldehyd -Lösung, Grade II, 25% in H2O
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Osmiumtetroxid -Lösung, suitable for electron microscopy, 4% in H2O
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Paraformaldehyd, reagent grade, crystalline
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Kaliumhexacyanoferrat(II) Trihydrat, ≥99.95% trace metals basis
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Monoklonales ANTI-Glucagon in Maus hergestellte Antikörper, clone K79bB10, ascites fluid
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Heparin Natriumsalz aus Schweinedarm, ≥180 USP units/mg
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Monoklonaler Anti-Insulin-Antikörper in Maus hergestellte Antikörper, clone K36AC10, ascites fluid
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Anti-SST antibody produced in rabbit, affinity isolated antibody, buffered aqueous glycerol solution
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Anti-Mouse IgG (whole molecule)–Gold antibody produced in goat, affinity isolated antibody, aqueous glycerol suspension, 10 nm (colloidal gold)
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Anti-Pancreatic polypeptide/PPY antibody produced in goat, affinity isolated antibody, buffered aqueous solution