Extracellular Electrophysiology on Clonal Human β-cell Spheroids

Overview

Journal Front Endocrinol (Lausanne)

Specialty Endocrinology

Date 2024 Jun 17

PMID 38883608

Authors

Emilie Puginier

Karen Leal-Fischer

Julien Gaitan

Marie Lallouet

Pier-Arnaldo Scotti

Matthieu Raoux

Jochen Lang

Affiliations

Soon will be listed here.

Abstract

Background: Pancreatic islets are important in nutrient homeostasis and improved cellular models of clonal origin may very useful especially in view of relatively scarce primary material. Close 3D contact and coupling between β-cells are a hallmark of physiological function improving signal/noise ratios. Extracellular electrophysiology using micro-electrode arrays (MEA) is technically far more accessible than single cell patch clamp, enables dynamic monitoring of electrical activity in 3D organoids and recorded multicellular slow potentials (SP) provide unbiased insight in cell-cell coupling.

Objective: We have therefore asked whether 3D spheroids enhance clonal β-cell function such as electrical activity and hormone secretion using human EndoC-βH1, EndoC-βH5 and rodent INS-1 832/13 cells.

Methods: Spheroids were formed either by hanging drop or proprietary devices. Extracellular electrophysiology was conducted using multi-electrode arrays with appropriate signal extraction and hormone secretion measured by ELISA.

Results: EndoC-βH1 spheroids exhibited increased signals in terms of SP frequency and especially amplitude as compared to monolayers and even single cell action potentials (AP) were quantifiable. Enhanced electrical signature in spheroids was accompanied by an increase in the glucose stimulated insulin secretion index. EndoC-βH5 monolayers and spheroids gave electrophysiological profiles similar to EndoC-βH1, except for a higher electrical activity at 3 mM glucose, and exhibited moreover a biphasic profile. Again, physiological concentrations of GLP-1 increased AP frequency. Spheroids also exhibited a higher secretion index. INS-1 cells did not form stable spheroids, but overexpression of connexin 36, required for cell-cell coupling, increased glucose responsiveness, dampened basal activity and consequently augmented the stimulation index.

Conclusion: In conclusion, spheroid formation enhances physiological function of the human clonal β-cell lines and these models may provide surrogates for primary islets in extracellular electrophysiology.

References

Tsonkova V, Sand F, Wolf X, Grunnet L, Ringgaard A, Ingvorsen C . The EndoC-βH1 cell line is a valid model of human beta cells and applicable for screenings to identify novel drug target candidates. Mol Metab. 2018; 8:144-157. PMC: 5985049. DOI: 10.1016/j.molmet.2017.12.007. View

Ravassard P, Hazhouz Y, Pechberty S, Bricout-Neveu E, Armanet M, Czernichow P . A genetically engineered human pancreatic β cell line exhibiting glucose-inducible insulin secretion. J Clin Invest. 2011; 121(9):3589-97. PMC: 3163974. DOI: 10.1172/JCI58447. View

Benninger R, Zhang M, Head W, Satin L, Piston D . Gap junction coupling and calcium waves in the pancreatic islet. Biophys J. 2008; 95(11):5048-61. PMC: 2586567. DOI: 10.1529/biophysj.108.140863. View

Roger B, Papin J, Vacher P, Raoux M, Mulot A, Dubois M . Adenylyl cyclase 8 is central to glucagon-like peptide 1 signalling and effects of chronically elevated glucose in rat and human pancreatic beta cells. Diabetologia. 2010; 54(2):390-402. DOI: 10.1007/s00125-010-1955-x. View

Wang Z, You J, Xu S, Hua Z, Zhang W, Deng T . Colocalization of insulin and glucagon in insulinoma cells and developing pancreatic endocrine cells. Biochem Biophys Res Commun. 2015; 461(4):598-604. DOI: 10.1016/j.bbrc.2015.04.072. View

Nlend R, Michon L, Bavamian S, Boucard N, Caille D, Cancela J . Connexin36 and pancreatic beta-cell functions. Arch Physiol Biochem. 2006; 112(2):74-81. DOI: 10.1080/13813450600712019. View

Nittala A, Wang X . The hyperbolic effect of density and strength of inter beta-cell coupling on islet bursting: a theoretical investigation. Theor Biol Med Model. 2008; 5:17. PMC: 2538510. DOI: 10.1186/1742-4682-5-17. View

Bollheimer L, Wrede C, Rockmann F, Ottinger I, Scholmerich J, Buettner R . Glucagon production of the rat insulinoma cell line INS-1-A quantitative comparison with primary rat pancreatic islets. Biochem Biophys Res Commun. 2005; 330(1):327-32. DOI: 10.1016/j.bbrc.2005.01.168. View

Rocheleau J, Remedi M, Granada B, Head W, Koster J, Nichols C . Critical role of gap junction coupled KATP channel activity for regulated insulin secretion. PLoS Biol. 2006; 4(2):e26. PMC: 1334237. DOI: 10.1371/journal.pbio.0040026. View

10.

Abarkan M, Pirog A, Mafilaza D, Pathak G, NKaoua G, Puginier E . Vertical Organic Electrochemical Transistors and Electronics for Low Amplitude Micro-Organ Signals. Adv Sci (Weinh). 2022; 9(8):e2105211. PMC: 8922095. DOI: 10.1002/advs.202105211. View

11.

Raoux M, Vacher P, Papin J, Picard A, Kostrzewa E, Devin A . Multilevel control of glucose homeostasis by adenylyl cyclase 8. Diabetologia. 2014; 58(4):749-57. DOI: 10.1007/s00125-014-3445-z. View

12.

Olcomendy L, Cassany L, Pirog A, Franco R, Puginier E, Jaffredo M . Towards the Integration of an Islet-Based Biosensor in Closed-Loop Therapies for Patients With Type 1 Diabetes. Front Endocrinol (Lausanne). 2022; 13:795225. PMC: 9072637. DOI: 10.3389/fendo.2022.795225. View

13.

Bosco D, Haefliger J, Meda P . Connexins: key mediators of endocrine function. Physiol Rev. 2011; 91(4):1393-445. DOI: 10.1152/physrev.00027.2010. View

14.

Krizhanovskii C, Kristinsson H, Elksnis A, Wang X, Gavali H, Bergsten P . EndoC-βH1 cells display increased sensitivity to sodium palmitate when cultured in DMEM/F12 medium. Islets. 2017; 9(3):e1296995. PMC: 5465947. DOI: 10.1080/19382014.2017.1296995. View

15.

Renaud S, Catargi B, Lang J . Biosensors in diabetes : how to get the most out of evolution and transpose it into a signal. IEEE Pulse. 2014; 5(3):30-4. DOI: 10.1109/MPUL.2014.2309577. View

16.

Ntamo Y, Samodien E, Burger J, Muller N, Muller C, Chellan N . Characterization of Insulin-Producing β-Cell Spheroids. Front Cell Dev Biol. 2021; 8:623889. PMC: 7876261. DOI: 10.3389/fcell.2020.623889. View

17.

Scharfmann R, Staels W, Albagli O . The supply chain of human pancreatic β cell lines. J Clin Invest. 2019; 129(9):3511-3520. PMC: 6715382. DOI: 10.1172/JCI129484. View

18.

Zbinden A, Urbanczyk M, Layland S, Becker L, Marzi J, Bosch M . Collagen and Endothelial Cell Coculture Improves β-Cell Functionality and Rescues Pancreatic Extracellular Matrix. Tissue Eng Part A. 2020; 27(13-14):977-991. DOI: 10.1089/ten.TEA.2020.0250. View

19.

Zbinden A, Marzi J, Schlunder K, Probst C, Urbanczyk M, Black S . Non-invasive marker-independent high content analysis of a microphysiological human pancreas-on-a-chip model. Matrix Biol. 2019; 85-86:205-220. DOI: 10.1016/j.matbio.2019.06.008. View

20.

Bugliani M, Syed F, Paula F, Omar B, Suleiman M, Mossuto S . DPP-4 is expressed in human pancreatic beta cells and its direct inhibition improves beta cell function and survival in type 2 diabetes. Mol Cell Endocrinol. 2018; 473:186-193. DOI: 10.1016/j.mce.2018.01.019. View