Preparation of Highly Functional Spheroid of Endocrine Cells Based on Thermosensitive Glycol Chitosan.

IF 4.4 4区 医学 Q2 CELL & TISSUE ENGINEERING
Seonmi Jang, Young-Woo Park, Kang Moo Huh, Dong Yun Lee
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引用次数: 0

Abstract

Background: Pancreatic islet transplantation holds great potential as a therapeutic approach for treating type 1 diabetes mellitus (T1D). However, large islets suffer from hypoxia due to the limited diffusion distance of oxygen, leading to cell loss. Therefore, smaller spheroids are needed for better transplantation outcomes. This study aims to develop a method for forming highly functional islet spheroids using glycol chitosan (GC) derivatives, such as N-acetylated glycol chitosan (AGC) and N-hexanoyl glycol chitosan (HGC).

Methods: Thermogelling polymers were produced by performing N-acylation of GC using the correspondingly carboxylic anhydrides. Islet spheroids were formed using a dual application with AGC-coated plates and HGC gelation. The AGC solution was applied to the plate for coating and evenly distributed using a 1 mL syringe. Then, the HGC encapsulated with islet single cells was cultured on top of it. Spheroid viability and functionality were evaluated using CCK-8 assay and glucose-stimulated insulin secretion assay.

Results: The aqueous solutions of AGC (4%, w/v) and HGC (36% hexanoylation) (2%, w/v) demonstrated a sol-gel transition temperature around 37 °C, suitable for the physiological environment. These polymers also showed no cytotoxicity to intact islets. Islet single cells were cultured on HGC gels with varying degrees of hexanoylation (DH) values, where higher DH values led to smaller and more uniform spheroids. The resulting spheroids formed on AGC-coated plates and HGC36 gelation were smaller and more uniform than those formed on untreated plates. These spheroids exhibited significantly improved glucose responsiveness, with superior insulin secretion.

Conclusion: The optimized method using AGC and HGC offers a more efficient way to produce smaller, uniform, and functional spheroids.

热敏乙二醇壳聚糖制备高功能内分泌细胞球体。
背景:胰岛移植作为治疗1型糖尿病(T1D)的一种治疗方法具有很大的潜力。然而,大的胰岛由于氧气的扩散距离有限而缺氧,导致细胞损失。因此,需要更小的球体以获得更好的移植效果。本研究旨在开发一种利用乙二醇壳聚糖(GC)衍生物,如n -乙酰化乙二醇壳聚糖(AGC)和n -己醇乙二醇壳聚糖(HGC)形成高功能胰岛球体的方法。方法:用相应的羧酸酐对GC进行n -酰化反应制备热凝胶聚合物。采用agc包被板和HGC凝胶双重应用形成胰岛球体。将AGC溶液涂于板上,用1ml注射器均匀分布。然后将胰岛单细胞包封的HGC置于其上培养。用CCK-8法和葡萄糖刺激胰岛素分泌法评估球体活力和功能。结果:AGC (4%, w/v)和HGC (36%, w/v)水溶液的溶胶-凝胶转变温度在37℃左右,适合生理环境。这些聚合物对完整的胰岛也没有细胞毒性。胰岛单细胞在不同程度己烯酰化(DH)值的HGC凝胶上培养,DH值越高,球体越小,越均匀。agc包覆板和HGC36凝胶形成的球体比未处理板形成的球体更小、更均匀。这些球状体表现出明显改善的葡萄糖反应性,具有优越的胰岛素分泌。结论:优化后的AGC - HGC法制备的球体体积更小、均匀、功能更强。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Tissue engineering and regenerative medicine
Tissue engineering and regenerative medicine CELL & TISSUE ENGINEERING-ENGINEERING, BIOMEDICAL
CiteScore
6.80
自引率
5.60%
发文量
83
审稿时长
6-12 weeks
期刊介绍: Tissue Engineering and Regenerative Medicine (Tissue Eng Regen Med, TERM), the official journal of the Korean Tissue Engineering and Regenerative Medicine Society, is a publication dedicated to providing research- based solutions to issues related to human diseases. This journal publishes articles that report substantial information and original findings on tissue engineering, medical biomaterials, cells therapy, stem cell biology and regenerative medicine.
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