Aggregation of Engineered Human β-Cells Into Pseudoislets: Insulin Secretion and Gene Expression Profile in Normoxic and Hypoxic Milieu.

M. Lecomte, Séverine Pechberty, C. Machado, Sandra Da Barroca, P. Ravassard, R. Scharfmann, P. Czernichow, B. Duvillié
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引用次数: 15

Abstract

Innovative treatments to cure type 1 diabetes are being actively researched. Among the different strategies, the replacement of β-cells has given promising results. Classically, islets from cadaveric donors are transplanted into diabetic patients, but recently phase I clinical trials that use stem cell-derived β-cells have been started. Such protocols require either an immunosuppressive treatment or the macroencapsulation of the β-cells. They involve cell aggregation and the exposure of the cells to hypoxia. Using an engineered human β-cell, we have addressed these two problems: a novel human β-cell line called EndoC-βH3 was cultured as single cells or aggregated clusters. EndoC-βH3 cells were also cultured at normal atmospheric oxygen tension (pO2 = 21%) or hypoxia (pO2 = 3%) in the presence or absence of modulators of the hypoxia-inducible factor 1α (HIF1α) pathway. Cell aggregation improved glucose-stimulated insulin secretion, demonstrating the benefit of cell-cell contacts. Low oxygen tension decreased β-cell viability and their sensitivity to glucose, but did not alter insulin production nor the insulin secretion capacity of the remaining cells. To investigate the role of HIF1α, we first used a HIF stabilizer at pO2 = 21%. This led to a mild decrease in cell viability, impaired glucose sensitivity, and altered insulin secretion. Finally, we used a HIF inhibitor on EndoC-βH3 pseudoislets exposed to hypoxia. Such treatment considerably decreased cell viability. In conclusion, aggregation of the EndoC-βH3 cells seems to be important to improve their function. A fraction of the EndoC-βH3 cells are resistant to hypoxia, depending on the level of activity of HIF1α. Thus, these cells represent a good human cell model for future investigations on islet cell transplantation analysis.
工程人β细胞聚集成假胰岛:常氧和低氧环境下胰岛素分泌和基因表达谱。
目前正在积极研究治疗1型糖尿病的创新疗法。在不同的策略中,β细胞的替代已经取得了可喜的结果。传统上,来自尸体供体的胰岛被移植到糖尿病患者身上,但最近,使用干细胞衍生的β细胞的I期临床试验已经开始。这种方案需要免疫抑制治疗或β细胞的大胶囊化。它们涉及细胞聚集和细胞暴露于缺氧。利用工程人类β细胞,我们解决了这两个问题:一种名为EndoC-βH3的新型人类β细胞系被培养为单细胞或聚集簇。EndoC-βH3细胞也在正常大气压(pO2 = 21%)或缺氧(pO2 = 3%)条件下,在存在或不存在缺氧诱导因子1α (HIF1α)通路调节剂的情况下培养。细胞聚集改善了葡萄糖刺激的胰岛素分泌,证明了细胞间接触的益处。低氧张力降低了β细胞的活力和对葡萄糖的敏感性,但没有改变剩余细胞的胰岛素产生和胰岛素分泌能力。为了研究HIF1α的作用,我们首先使用pO2 = 21%的HIF稳定剂。这导致细胞活力轻度下降,葡萄糖敏感性受损,胰岛素分泌改变。最后,我们将HIF抑制剂应用于缺氧条件下的EndoC-βH3假胰岛。这种处理大大降低了细胞活力。综上所述,EndoC-βH3细胞的聚集似乎对改善其功能很重要。一部分EndoC-βH3细胞对缺氧具有抗性,这取决于HIF1α的活性水平。因此,这些细胞为未来胰岛细胞移植分析研究提供了良好的人类细胞模型。
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来源期刊
Cell medicine
Cell medicine MEDICINE, RESEARCH & EXPERIMENTAL-
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