Evaluation of morphology and viability of spheroid derived from Insulin-GLase cell line: A model system to understand Type 2 Diabetes Mellitus

Abstract

Type 2 Diabetes Mellitus (T2DM) is one of the major health issues in the world. The cellular mechanism of T2DM is still not fully understood. It could be studied by using spheroid three-dimensional (3D) culture which is considered representative of the in vivo conditions. Several types of pancreatic β cell lines have been used, one of which is the insulin-GLase (iGL) cell line. This study aims to evaluate the effect of cell density and incubation time on spheroid morphology and cell viability in order to understand which one can be considered as the best option in studying T2DM using iGL cell. Spheroid was made by using the Hanging drop method. The variations of initial seeding cells were 50, 100, 200, and 400 cells/µL then incubated for 1, 2, 3, and 4 days. The evaluated parameters in this study are spheroid morphology and cell viability. Spheroid morphology was observed by using inverted phase contrast microscope integrated with camera (Nikon) and NIS-Elements Analysis D software. Cell viability was determined by using LUNA-II™ Automated Cell Counter (Logos Biosystem). The result of this study showed that spheroid in all of the group cell concentration have formed since the first day and its diameter was significantly increased on the following days (p<0,05). The spheroid size was positively correlated with the cell density in group 50-200 cells/µL. A single and stable spheroid morphology was observed in 50-100 cells/µL group. Cell viability in 3D culture system was lower and significantly decreased since day 3 compared to 2D culture (p <0.05; 0.01). In conclusion, spheroid derived from iGL cell line with a stable morphology and good viability could be obtained from a cell concentration of 50-100 cells / µL with two days of incubation.