Human Cell Design, Canceropole, Toulouse, France.
Human Cell Design, Canceropole, Toulouse, France.
Mol Metab. 2023 Oct;76:101772. doi: 10.1016/j.molmet.2023.101772. Epub 2023 Jul 11.
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.
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.
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.
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.
易于获得且功能上接近成人胰岛β细胞的人类胰岛β细胞是更好地了解人类β细胞生理学和开发新的糖尿病治疗方法的关键模型。我们在此报告 EndoC-βH5 细胞的特征，这是 EndoC-βH 细胞系的最新成员。
通过将永生化转基因 hTERT 和 SV40 大 T 以及单纯疱疹病毒-1 胸苷激酶整合基因转移到人胎胰腺中，生成 EndoC-βH5 细胞。使用 CRE 激活扩增后去除永生化转基因，并用更昔洛韦去除未切除的细胞。得到的细胞被分配为即用型 EndoC-βH5 细胞。我们进行了转录组、免疫学和广泛的功能测定。
即用型 EndoC-βH5 细胞表现出高效的葡萄糖依赖性胰岛素分泌。在欧洲的四个独立实验室中观察到并复制了强大的 10 倍胰岛素分泌指数。EndoC-βH5 细胞以动态方式响应葡萄糖分泌胰岛素，并且 GIP 和 GLP-1 类似物进一步增强了分泌。RNA-seq 证实了丰富的β细胞转录因子和功能标志物的表达，包括肠促胰岛素受体。细胞因子诱导炎症途径和抗原加工和呈递的基因表达特征。最后，表达修饰的 HLA-A2 的 EndoC-βH5 细胞引发特异性 A2 同种反应性 CD8 T 细胞激活。
EndoC-βH5 细胞代表了一种独特的可储存和即用型人类胰岛β细胞模型，具有高度稳健和可重复的特征。因此，这些细胞与β细胞功能研究、新药物的筛选和验证以及疾病模型的发展相关。
