UNASSIGNED: 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.
UNASSIGNED: 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.
UNASSIGNED: 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.
UNASSIGNED: 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.
■因此,我们询问3D球体是否使用人EndoC-βH1,EndoC-βH5和啮齿动物INS-1832/13细胞增强克隆β细胞功能,例如电活性和激素分泌。
■球体是通过悬挂式或专有设备形成的。使用多电极阵列进行细胞外电生理学,并通过ELISA测量适当的信号提取和激素分泌。
■与单层相比,EndoC-βH1球体在SP频率和尤其是振幅方面表现出增加的信号,甚至单细胞动作电位(AP)也是可量化的。球状体中增强的电特征伴随着葡萄糖刺激的胰岛素分泌指数的增加。EndoC-βH5单层和球体的电生理特性与EndoC-βH1相似,但在3mM葡萄糖下具有更高的电活性,此外,还表现出双相的轮廓。再一次,GLP-1的生理浓度增加AP频率。球体也表现出更高的分泌指数。INS-1细胞没有形成稳定的球体,但是细胞-细胞偶联所需的连接蛋白36的过表达,葡萄糖反应性增加,抑制了基础活动,因此增加了刺激指数。
■总而言之,球体的形成增强了人克隆β细胞系的生理功能,这些模型可能为细胞外电生理学中的原代胰岛提供替代。