title: Acinar phenotype is preserved in human exocrine pancreas cells cultured at low temperature: implications for lineage-tracing of β-cell neogenesis.
abstract: The regenerative medicine field is expanding with great successes in laboratory and preclinical settings. Pancreatic acinar cells in diabetic mice were recently converted into β-cells by treatment with ciliary neurotrophic factor (CNTF) and epidermal growth factor (EGF). This suggests that human acinar cells might become a cornerstone for diabetes cell therapy in the future, if they can also be converted into glucose-responsive insulin-producing cells. Presently, studying pancreatic acinar cell biology in vitro is limited by their high plasticity, as they rapidly lose their phenotype and spontaneously transdifferentiate to a duct-like phenotype in culture. We questioned whether human pancreatic acinar cell phenotype could be preserved in vitro by physico-chemical manipulations and whether this could be valuable in the study of β-cell neogenesis. We found that culture at low temperature (4°C) resulted in the maintenance of morphological and molecular acinar cell characteristics. Specifically, chilled acinar cells did not form the spherical clusters observed in controls (culture at 37°C), and they maintained high levels of acinar-specific transcripts and proteins. Five-day chilled acinar cells still transdifferentiated into duct-like cells upon transfer to 37°C. Moreover, adenoviral-mediated gene transfer evidenced an active Amylase promoter in the 7-day chilled acinar cells, and transduction performed in chilled conditions improved acinar cell labelling. Together, our findings indicate the maintenance of human pancreatic acinar cell phenotype at low temperature and the possibility to efficiently label acinar cells, which opens new perspectives for the study of human acinar-to-β-cell transdifferentiation.
authors.affiliation: Cell Differentiation Unit, Diabetes Research Center, Vrije Universiteit Brussel (VUB), Brussels, BE 1090, Belgium.
authors.forename: Josué K
authors.initials: JK
authors.lastname: Mfopou
authors.affiliation: Cell Differentiation Unit, Diabetes Research Center, Vrije Universiteit Brussel (VUB), Brussels, BE 1090, Belgium.
authors.forename: Isabelle
authors.initials: I
authors.lastname: Houbracken
authors.affiliation: Cell Differentiation Unit, Diabetes Research Center, Vrije Universiteit Brussel (VUB), Brussels, BE 1090, Belgium.
authors.forename: Elke
authors.initials: E
authors.lastname: Wauters
authors.affiliation: Cell Differentiation Unit, Diabetes Research Center, Vrije Universiteit Brussel (VUB), Brussels, BE 1090, Belgium.
authors.forename: Iris
authors.initials: I
authors.lastname: Mathijs
authors.affiliation: Cell Differentiation Unit, Diabetes Research Center, Vrije Universiteit Brussel (VUB), Brussels, BE 1090, Belgium.
authors.forename: Imane
authors.initials: I
authors.lastname: Song
authors.affiliation: Cell Differentiation Unit, Diabetes Research Center, Vrije Universiteit Brussel (VUB), Brussels, BE 1090, Belgium.
authors.forename: Eddy
authors.initials: E
authors.lastname: Himpe
authors.affiliation: Cell Differentiation Unit, Diabetes Research Center, Vrije Universiteit Brussel (VUB), Brussels, BE 1090, Belgium.
authors.forename: Jonathan
authors.initials: J
authors.lastname: Baldan
authors.affiliation: Beta Cell Neogenesis, Diabetes Research Center, Vrije Universiteit Brussel (VUB), Brussels, BE 1090, Belgium.
authors.forename: Harry
authors.initials: H
authors.lastname: Heimberg
authors.affiliation: Cell Differentiation Unit, Diabetes Research Center, Vrije Universiteit Brussel (VUB), Brussels, BE 1090, Belgium lucbo@vub.ac.be.
authors.forename: Luc
authors.initials: L
authors.lastname: Bouwens
mesh_terms: D000681:Amylases / Q000235:genetics; D000818:Animals; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D019070:Cell Lineage; D054338:Cell Transdifferentiation; D002478:Cells, Cultured; D003080:Cold Temperature; D006801:Humans; D050417:Insulin-Secreting Cells / Q000166:cytology* / Q000378:metabolism; D051379:Mice; D046790:Pancreas, Exocrine / Q000166:cytology* / Q000378:metabolism; D010641:Phenotype; D011401:Promoter Regions, Genetic; D059467:Transcriptome
chemical_list: D000681:Amylases
keywords: acinar cell; chilled; hypothermic; lineage tracing; neogenesis; transdifferentiation; β-cell