Generation of β-like cell subtypes from differentiated human induced pluripotent stem cells in 3D spheroids - Archive ouverte HAL Accéder directement au contenu
Article Dans Une Revue Molecular Omics Année : 2023

Generation of β-like cell subtypes from differentiated human induced pluripotent stem cells in 3D spheroids

Stéphane Poulain
Soo Hyeon Kim
Cecile Legallais
Rachid Jellali

Résumé

Since the identification of four different pancreatic β-cell subtypes and bi-hormomal cells playing a role in the diabetes pathogenesis, the search for in vitro models that mimics such cells heterogeneity became a key priority in experimental and clinical diabetology. We investigated the potential of human induced pluripotent stem cells to lead to the development of the different β-cells subtypes in honeycomb microwell-based 3D spheroids. The glucose-stimulated insulin secretion confirmed the spheroids functionality. Then, we performed a single cell RNA sequencing of the spheroids. Using a knowledge-based analysis with a stringency on the pancreatic markers, we extracted the β-cells INS+/UCN3+ subtype (11%; β1-like cells), the INS+/ST8SIA1+/CD9− subtype (3%, β3-like cells) and INS+/CD9+/ST8SIA1-subtype (1%; β2-like cells) consistently with literature findings. We did not detect the INS+/ST8SIA1+/CD9+ cells (β4-like cells). Then, we also identified four bi-hormonal cells subpopulations including δ-like cells (INS+/SST+, 6%), γ-like cells (INS+/PPY+, 3%), α-like-cells (INS+/GCG+, 6%) and ε-like-cells (INS+/GHRL+, 2%). Using data-driven clustering, we extracted four progenitors’ subpopulations (with the lower level of INS gene) that included one population highly expressing inhibin genes (INHBA+/INHBB+), one population highly expressing KCNJ3+/TPH1+, one population expressing hepatocyte-like lineage markers (HNF1A+/AFP+), and one population expressing stem-like cell pancreatic progenitor markers (SOX2+/NEUROG3+). Furthermore, among the cycling population we found a large number of REST+ cells and CD9+ cells (CD9+/SPARC+/REST+). Our data confirm that our differentiation leads to large β-cell heterogeneity, which can be used for investigating β-cells plasticity under physiological and pathophysiological conditions.
Fichier principal
Vignette du fichier
Mol_Omics_AAM.pdf (5.45 Mo) Télécharger le fichier
Origine : Fichiers produits par l'(les) auteur(s)

Dates et versions

hal-04209565 , version 1 (06-11-2023)

Identifiants

Citer

Lisa Morisseau, Fumiya Tokito, Stéphane Poulain, Valérie Plaisance, Valerie Pawlowski, et al.. Generation of β-like cell subtypes from differentiated human induced pluripotent stem cells in 3D spheroids. Molecular Omics, 2023, 19 (10), pp.810-822. ⟨10.1039/D3MO00050H⟩. ⟨hal-04209565⟩
34 Consultations
1 Téléchargements

Altmetric

Partager

Gmail Facebook X LinkedIn More