EndoC-βH5 cells are storable and ready-to-use human pancreatic beta cells with physiological insulin secretion
(2023) In Molecular Metabolism 76.- Abstract
Objectives: Readily accessible human pancreatic beta cells that are functionally close to primary adult beta cells are a crucial model to better understand human beta cell physiology and develop new treatments for diabetes. We here report the characterization of EndoC-βH5 cells, the latest in the EndoC-βH cell family. Methods: EndoC-βH5 cells were generated by integrative gene transfer of immortalizing transgenes hTERT and SV40 large T along with Herpes Simplex Virus-1 thymidine kinase into human fetal pancreas. Immortalizing transgenes were removed after amplification using CRE activation and remaining non-excized cells eliminated using ganciclovir. Resulting cells were distributed as ready to use EndoC-βH5 cells. We performed... (More)
Objectives: Readily accessible human pancreatic beta cells that are functionally close to primary adult beta cells are a crucial model to better understand human beta cell physiology and develop new treatments for diabetes. We here report the characterization of EndoC-βH5 cells, the latest in the EndoC-βH cell family. Methods: EndoC-βH5 cells were generated by integrative gene transfer of immortalizing transgenes hTERT and SV40 large T along with Herpes Simplex Virus-1 thymidine kinase into human fetal pancreas. Immortalizing transgenes were removed after amplification using CRE activation and remaining non-excized cells eliminated using ganciclovir. Resulting cells were distributed as ready to use EndoC-βH5 cells. We performed transcriptome, immunological and extensive functional assays. Results: Ready to use EndoC-βH5 cells display highly efficient glucose dependent insulin secretion. A robust 10-fold insulin secretion index was observed and reproduced in four independent laboratories across Europe. EndoC-βH5 cells secrete insulin in a dynamic manner in response to glucose and secretion is further potentiated by GIP and GLP-1 analogs. RNA-seq confirmed abundant expression of beta cell transcription factors and functional markers, including incretin receptors. Cytokines induce a gene expression signature of inflammatory pathways and antigen processing and presentation. Finally, modified HLA-A2 expressing EndoC-βH5 cells elicit specific A2-alloreactive CD8 T cell activation. Conclusions: EndoC-βH5 cells represent a unique storable and ready to use human pancreatic beta cell model with highly robust and reproducible features. Such cells are thus relevant for the study of beta cell function, screening and validation of new drugs, and development of disease models.
(Less)
- author
- Blanchi, Bruno
; Taurand, Marion
; Colace, Claire
; Thomaidou, Sofia
; Audeoud, Charlotte
; Fantuzzi, Federica
; Sawatani, Toshiaki
; Gheibi, Sevda
LU
; Sabadell-Basallote, Joan
; Boot, Fransje W.J.
; Chantier, Thibault
; Piet, Aline
; Cavanihac, Charlotte
; Pilette, Marion
; Balguerie, Adélie
; Olleik, Hamza
; Carlotti, Françoise
; Ejarque, Miriam
; Fex, Malin
LU
; Mulder, Hindrik
LU
; Cnop, Miriam
; Eizirik, Decio L.
; Jouannot, Ouardane
; Gaffuri, Anne Lise
; Czernichow, Paul
; Zaldumbide, Arnaud
; Scharfmann, Raphaël
and Ravassard, Philippe
(Less) - organization
- publishing date
- 2023-10
- type
- Contribution to journal
- publication status
- published
- subject
- keywords
- Glucose and incretin stimulated insulin secretion, Human beta cell function, Human pancreatic beta cell line, Type-I diabetes disease model
- in
- Molecular Metabolism
- volume
- 76
- article number
- 101772
- publisher
- Elsevier
- external identifiers
-
- pmid:37442376
- scopus:85166595030
- ISSN
- 2212-8778
- DOI
- 10.1016/j.molmet.2023.101772
- language
- English
- LU publication?
- yes
- id
- 01a5b03e-af9d-4e0a-9e78-561c1ebdbeff
- date added to LUP
- 2023-10-24 14:25:01
- date last changed
- 2024-04-19 03:51:03
@article{01a5b03e-af9d-4e0a-9e78-561c1ebdbeff,
abstract = {{<p>Objectives: Readily accessible human pancreatic beta cells that are functionally close to primary adult beta cells are a crucial model to better understand human beta cell physiology and develop new treatments for diabetes. We here report the characterization of EndoC-βH5 cells, the latest in the EndoC-βH cell family. Methods: EndoC-βH5 cells were generated by integrative gene transfer of immortalizing transgenes hTERT and SV40 large T along with Herpes Simplex Virus-1 thymidine kinase into human fetal pancreas. Immortalizing transgenes were removed after amplification using CRE activation and remaining non-excized cells eliminated using ganciclovir. Resulting cells were distributed as ready to use EndoC-βH5 cells. We performed transcriptome, immunological and extensive functional assays. Results: Ready to use EndoC-βH5 cells display highly efficient glucose dependent insulin secretion. A robust 10-fold insulin secretion index was observed and reproduced in four independent laboratories across Europe. EndoC-βH5 cells secrete insulin in a dynamic manner in response to glucose and secretion is further potentiated by GIP and GLP-1 analogs. RNA-seq confirmed abundant expression of beta cell transcription factors and functional markers, including incretin receptors. Cytokines induce a gene expression signature of inflammatory pathways and antigen processing and presentation. Finally, modified HLA-A2 expressing EndoC-βH5 cells elicit specific A2-alloreactive CD8 T cell activation. Conclusions: EndoC-βH5 cells represent a unique storable and ready to use human pancreatic beta cell model with highly robust and reproducible features. Such cells are thus relevant for the study of beta cell function, screening and validation of new drugs, and development of disease models.</p>}},
author = {{Blanchi, Bruno and Taurand, Marion and Colace, Claire and Thomaidou, Sofia and Audeoud, Charlotte and Fantuzzi, Federica and Sawatani, Toshiaki and Gheibi, Sevda and Sabadell-Basallote, Joan and Boot, Fransje W.J. and Chantier, Thibault and Piet, Aline and Cavanihac, Charlotte and Pilette, Marion and Balguerie, Adélie and Olleik, Hamza and Carlotti, Françoise and Ejarque, Miriam and Fex, Malin and Mulder, Hindrik and Cnop, Miriam and Eizirik, Decio L. and Jouannot, Ouardane and Gaffuri, Anne Lise and Czernichow, Paul and Zaldumbide, Arnaud and Scharfmann, Raphaël and Ravassard, Philippe}},
issn = {{2212-8778}},
keywords = {{Glucose and incretin stimulated insulin secretion; Human beta cell function; Human pancreatic beta cell line; Type-I diabetes disease model}},
language = {{eng}},
publisher = {{Elsevier}},
series = {{Molecular Metabolism}},
title = {{EndoC-βH5 cells are storable and ready-to-use human pancreatic beta cells with physiological insulin secretion}},
url = {{http://dx.doi.org/10.1016/j.molmet.2023.101772}},
doi = {{10.1016/j.molmet.2023.101772}},
volume = {{76}},
year = {{2023}},
}