Simulated Cholinergic Reinnervation of β (INS-1) Cells: Antidiabetic Utility of Heterotypic Pseudoislets Containing β Cell and Cholinergic Cell.

Ao Jiao,Feng Li,Chengshuo Zhang,Wu Lv,Jialin Zhang,Baomin Chen

doi:10.1155/2018/1505307

Ao Jiao, Feng Li + Show 4 more

Open Access

https://doi.org/10.1155/2018/1505307

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Abstract

Cholinergic neurons can functionally support pancreatic islets in controlling blood sugar levels. However, in islet transplantation, the level of cholinergic reinnervation is significantly lower compared to orthotopic pancreatic islets. This abnormal reinnervation affects the survival and function of islet grafts. In this study, the cholinergic reinnervation of beta cells was simulated by 2D and 3D coculture of INS-1 and NG108-15 cells. In 2D culture conditions, 20 mM glucose induced a 1.24-fold increase (p < 0.0001) in insulin secretion from the coculture group, while in the 3D culture condition, a 1.78-fold increase (p < 0.0001) in insulin secretion from heterotypic pseudoislet group was observed. Glucose-stimulated insulin secretion (GSIS) from 2D INS-1 cells showed minimal changes when compared to 3D structures. E-cadherin expressed in INS-1 and NG108-15 cells was the key adhesion molecule for the formation of heterotypic pseudoislets. NG108-15 cells hardly affected the proliferation of INS-1 cells in vitro. Heterotypic pseudoislet transplantation recipient mice reverted to normoglycemic levels faster and had a greater blood glucose clearance compared to INS-1 pseudoislet recipient mice. In conclusion, cholinergic cells can promote insulin-secreting cells to function better in vitro and in vivo and E-cadherin plays an important role in the formation of heterotypic pseudoislets.

Highlights

Islet transplantation is a beneficial approach for the treatment of type 1 diabetes (T1DM)
The glucose-stimulated insulin secretion (GSIS) results of 2D coculture confirmed our hypothesis that cholinergic NG10815 cells enhanced the Glucose-stimulated insulin secretion (GSIS) capacity of INS-1 β cells
The GSIS capacity of these heterotypic pseudoislets was enhanced, with insulin release nearly doubling with high glucose (20 mM) stimulation

Summary

Introduction

Islet transplantation is a beneficial approach for the treatment of type 1 diabetes (T1DM). INS-1 cells are widely used as rat islet β cell models for diabetes research They express M1 and M3 receptors, which are activated by carbachol to promote insulin release [9]. The NG108-15 cell line has the ability to release acetylcholine and was created by fusing mouse N18TG2 neuroblastoma cells with rat C6-BU-1 glioma cells in the presence of inactivated Sendai virus [11]. This cell line is frequently used as a cholinergic cell line to explore neuronal functions [12]. Therapeutic potential of heterotypic pseudoislets generated from INS-1 β cells and the cholinergic cell line NG108-15 was examined.

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