title: Aggregation of Engineered Human β-Cells Into Pseudoislets: Insulin Secretion and Gene Expression Profile in Normoxic and Hypoxic Milieu.
abstract: Innovative treatments to cure type 1 diabetes are being actively researched. Among the different strategies, the replacement of β-cells has given promising results. Classically, islets from cadaveric donors are transplanted into diabetic patients, but recently phase I clinical trials that use stem cell-derived β-cells have been started. Such protocols require either an immunosuppressive treatment or the macroencapsulation of the β-cells. They involve cell aggregation and the exposure of the cells to hypoxia. Using an engineered human β-cell, we have addressed these two problems: a novel human β-cell line called EndoC-βH3 was cultured as single cells or aggregated clusters. EndoC-βH3 cells were also cultured at normal atmospheric oxygen tension (pO2= 21%) or hypoxia (pO2= 3%) in the presence or absence of modulators of the hypoxia-inducible factor 1α (HIF1α) pathway. Cell aggregation improved glucose-stimulated insulin secretion, demonstrating the benefit of cell-cell contacts. Low oxygen tension decreased β-cell viability and their sensitivity to glucose, but did not alter insulin production nor the insulin secretion capacity of the remaining cells. To investigate the role of HIF1α, we first used a HIF stabilizer at pO2= 21%. This led to a mild decrease in cell viability, impaired glucose sensitivity, and altered insulin secretion. Finally, we used a HIF inhibitor on EndoC-βH3 pseudoislets exposed to hypoxia. Such treatment considerably decreased cell viability. In conclusion, aggregation of the EndoC-βH3 cells seems to be important to improve their function. A fraction of the EndoC-βH3 cells are resistant to hypoxia, depending on the level of activity of HIF1α. Thus, these cells represent a good human cell model for future investigations on islet cell transplantation analysis.
authors.affiliation: Univercell-Biosolutions, Centre de recherche des Cordeliers , Paris , France.
authors.forename: Marie-José
authors.initials: MJ
authors.lastname: Lecomte
authors.affiliation: Univercell-Biosolutions, Centre de recherche des Cordeliers , Paris , France.
authors.forename: Séverine
authors.initials: S
authors.lastname: Pechberty
authors.affiliation: Univercell-Biosolutions, Centre de recherche des Cordeliers , Paris , France.
authors.forename: Cécile
authors.initials: C
authors.lastname: Machado
authors.affiliation: Univercell-Biosolutions, Centre de recherche des Cordeliers , Paris , France.
authors.forename: Sandra
authors.initials: S
authors.lastname: Da Barroca
authors.affiliation: † Sorbonne Universités, UPMC Univ Paris 06, Inserm, CNRS, Institut du cerveau et de la moelle (ICM)-Hôpital Pitié-Salpêtrière , Paris , France.
authors.forename: Philippe
authors.initials: P
authors.lastname: Ravassard
authors.affiliation: ‡INSERM U1016, Institut Cochin, Paris, France; §Université Paris Descartes, Sorbonne Paris Cité, Faculté de Médecine, Paris, France.
authors.forename: Raphaël
authors.initials: R
authors.lastname: Scharfmann
authors.affiliation: Univercell-Biosolutions, Centre de recherche des Cordeliers , Paris , France.
authors.forename: Paul
authors.initials: P
authors.lastname: Czernichow
authors.affiliation: ‡INSERM U1016, Institut Cochin, Paris, France; §Université Paris Descartes, Sorbonne Paris Cité, Faculté de Médecine, Paris, France.
authors.forename: Bertrand
authors.initials: B
authors.lastname: Duvillié
keywords: HIF1α; Hypoxia; Insulin secretion; Pseudoislets (PIs); β-Cells