EndoC-βH5 cells are storable and ready-to-use human pancreatic beta cells with physiological insulin secretion

Blanchi, Bruno; Taurand, Marion; Colace, Claire; Thomaidou, Sofia; Audeoud, Charlotte; Fantuzzi, Federica; Sawatani, Toshiaki; Gheibi, Sevda; 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; Mulder, Hindrik; Cnop, Miriam; Eizirik, Decio L.; Jouannot, Ouardane; Gaffuri, Anne-Lise; Czernichow, Paul; Zaldumbide, Arnaud; Scharfmann, Raphaël; Ravassard, Philippe

doi:10.1016/j.molmet.2023.101772

Cited by 5 publications

(9 citation statements)

References 48 publications

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“…Immunofluorescence demonstrated the expected presence of insulin as well as the SNARE proteins VAMP2 and SNAP-25. As reported previously (9), EndoC-βH5 also stained for glucagon and somatostatin in spheroids (Supplemental Fig. 2) as did monolayers (data not shown).…”

Section: Resultssupporting

confidence: 88%

“…We subsequently tested an optimized EndoC-βH version (Fig. 3), the EndoC-βH5 cells, known for their improved function (9). EndoC-βH5 easily formed stable spheroids (Supplemental Fig.…”

Section: Resultsmentioning

confidence: 99%

“…Studies on primary islet cells are hampered by the relative scarceness of native material, especially those of human origin, which moreover differ in several important aspects from rodent islet cells (3)and exhibit a high degree of variability (4). Consequently, clonal β-cell lines provide useful models and this approach has been considerably improved by the establishment of a human β-cell line, EndoC-βH1 cells and their derivatives (5-9).…”

Section: Introductionmentioning

confidence: 99%

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Extracellular electrophysiology on clonal human β-cell spheroids

Puginier,

Fischer,

Gaitan

et al. 2024

Preprint

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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 beta-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. We have therefore asked whether 3D spheroids enhance clonal beta-cell function such as electrical activity and hormone secretion using human EndoC- betaH1, EndoC- betaH5 and rodent INS-1 cells. EndoC- betaH1 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- betaH5 monolayers and spheroids gave electrophysiological profiles similar to EndoC- betaH1, 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. In conclusion, spheroid formation enhances physiological function of the human clonal beta-cell lines and these models may provide surrogates for primary islets in extracellular electrophysiology.

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Section: Resultssupporting

confidence: 88%

“…We subsequently tested an optimized EndoC-βH version (Fig. 3), the EndoC-βH5 cells, known for their improved function (9). EndoC-βH5 easily formed stable spheroids (Supplemental Fig.…”

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Extracellular electrophysiology on clonal human β-cell spheroids

Puginier,

Fischer,

Gaitan

et al. 2024

Preprint

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“…At this concentration, AXKO-0046 suppressed both glucose-stimulated ATP/ADP ratios and Ca 2+ fluxes (Figure S3D-G). Analysis of transcriptomic data revealed that EndoC-βH5 spheroids express moderate LDHA levels, which might explain the exaggerated effects of LDHB inhibition versus primary human β cells (64.11 ± 2.60 versus 247.73 ± 5.00 TPM, LDHA versus LDHB ; mean ± SD; taken from GSE224732 48 ).…”

Section: Resultsmentioning

confidence: 99%

High-resolution glucose tracing and in situ imaging reveals regulated lactate production in human pancreatic β cells

Cuozzo

Nasteska

Jiao

et al. 2022

Preprint

Self Cite

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Using 13C6 glucose labeling coupled to GC-MS and 2D 1H-13C HSQC NMR spectroscopy, we have obtained a comparative high-resolution map of glucose fate underpinning steady state insulin release and β cell function. In both mouse and human islets, the contribution of glucose to the TCA cycle is similar. Pyruvate-fueling of the TCA cycle is found to be mediated primarily by the activity of pyruvate dehydrogenase (PDH), with only a limited contribution from pyruvate carboxylase (PC). While conversion of pyruvate to lactate by lactate dehydrogenase (LDH) can be detected in both species, lactate accumulation via this route is six-fold higher in human islets. Transcriptomic analysis reveals that human β cells specifically express lactate dehydrogenase B (LDHB) at high levels, in keeping with the phenotype of patients harboring gain-of-function mutations in MCT1/ SLC16A1 (HHF7). Thus, glycolytically-derived acetyl CoA preferentially feeds the TCA cycle in both mouse and human β cells. However, human β cells possess the machinery needed to generate extra-mitochondrial lactate, which might reflect a key mechanism to balance the reducing activity of NADH-producing pathways.

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“…[ 42 ] Recently, it was reported that βH5 cells engineered to express HLA‐A2 (human leukocyte antigen‐A2) can activate cytotoxic T cells in a co‐culture model, providing a valuable tool to study T cell interactions with human β‐cells. [ 43 ] These advances provide more versatility in selecting accurate models for researchers to answer their research questions.…”

Section: In Vitro and In Vivo Models Of T1dmentioning

confidence: 99%

Advances in the Use of Biologics and Biomaterials toward the Improvement of Pancreatic Islet Graft Survival in Type 1 Diabetes

Yitayew,

Luginina,

Tabrizian

2024

Advanced NanoBiomed Research

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Islet transplantation is a curative treatment for patients suffering from type 1 diabetes and has the potential to replace current treatment strategies involving the exogenous administration of insulin. Despite this potential, there are many hurdles in achieving successful long‐term graft survival due to autoimmune and foreign body reactions leading to graft rejection coupled with donor shortage and potential adverse effects from the need for long‐term administration of immunosuppressive drugs. As a result, various approaches have been proposed to increase the viability and function of islet grafts during isolation and ex vivo culture with the use of growth factors, hormones, and other therapeutic agents. In addition, other strategies have addressed how to enhance or maintain islet graft performance after implantation with improvements on immunosuppressive drug regimens and the use of biomaterials to encapsulate and protect the cells from graft rejection. This review focuses on the recent advances in strategies to improve islet viability and function with the addition of exogenous compounds and the implementation of conformal coating as a promising tool for immunoprotection of islet transplants.

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EndoC-βH5 cells are storable and ready-to-use human pancreatic beta cells with physiological insulin secretion

Cited by 5 publications

References 48 publications

Extracellular electrophysiology on clonal human β-cell spheroids

Extracellular electrophysiology on clonal human β-cell spheroids

High-resolution glucose tracing and in situ imaging reveals regulated lactate production in human pancreatic β cells

Advances in the Use of Biologics and Biomaterials toward the Improvement of Pancreatic Islet Graft Survival in Type 1 Diabetes

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