Nanoencapsulation with T3 loaded polymersomes improves the function of neonatal porcine islet like cell clusters
Sang Hoon Lee1,2, Min Se Kim2, Eun Jin Lee1, Sun Mi Ahn1, Hyun-Ouk Kim2, Jung-Taek Kang1.
1Biotechnology Research Institute, Mgensolution, Seoul, Korea; 2Department of Biotechnology and Bioengineering, Kangwon National University, Chuncheon, Korea
Introduction: Neonatal porcine islet like cell clusters (NPCCs) can be used to as a resource to treat type Ⅰ diabetes. To overcome a xenogenic immunogenicity of the NPCCs, nanoencapsulation of NPCCs have been tried. Previously, we confirmed the effects of immune protection in rodent models through nanoencapsulation of NPCCs using polyethylene glycol-b-polylactic acid based polymersomes (PSomes). In this study, to increase binding efficiency of PSomes on NPCCs, PSome surface was modified with N-hydroxysuccinimide (NHS) and Maleimide (Mal), in which can induce dual covalent bond for immunoisolation by binding the amine and thiol groups on NPCCs, respectively. Moreover, the triiodothyronine (T3) concerned with maturation of pancreatic β cells and islets differentiation derived stem cell, was loaded in PSomes. We evaluated whether T3 loaded PSomes improve the efficiency of nano-encapsulation and functionality of NPCCs.
Methods: NPCCs were isolated from neonatal porcine pancreas using enzyme digestion method and cultured during 8 days. And then, NPCCs were nanoencapsulated with T3-PSomes. The efficiency was measured by mean fluorescence intensity (MFI) of DiI using either fluorescence microscopy or confocal laser scanning microscopy. The functional assessment of nanoencapsulated NPCCs with T3-PSomes was evaluated by GSIS assay and qRT-PCR.
Results: The mean diameter of PSomes measured by dynamic light scattering showed that PSomes encapsulate conformally the surface of NPCCs and the MFI increases time-dependently. Nanoencapsulated with T3-PSomes didn’t affects the viability of NPCCs. Although the insulin secretion at high concentrations of glucose stimulation didn’t show a significant difference in the T3-PSome group compared with control group, insulin mRNA expression and normal insulin secretion were increased significantly in T3-PSome group, compared with control group. However, there was no significant difference in stimulation index among all groups.
Conclusions: Our study will provide the improved immunoisolation strategy through efficient nanoencapsulation of NPCCs through induction of dual covalent bond and will overcome the functional immaturity by stimulating differentiation of the NPCCs through a constant release of T3 loaded in PSomes. Also, our loading system will offer opportunity to use various maturation related chemical and functional reagent. Therefore, we expect that NPCCS can be used as the optimal source for islet xenotransplantation. Further studies are needed to demonstrate the immune protection effects of our nanoencapsulated NPCCs in human immune system.
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