构建胰岛细胞片的人层粘连蛋白同型涂层。

Shingo Yamashita, K. Ohashi, R. Utoh, T. Okano, Masakazu Yamamoto
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引用次数: 15

摘要

我们开发新的基于胰岛的糖尿病治疗方法的实验方法是创建单层胰岛细胞结构(胰岛细胞片),然后将其移植到皮下口袋中。先前的研究将大鼠层粘胶蛋白-5(链组成:α3, β3, γ2)描述为一种合适的细胞外基质(ECM),用于由涂层温度响应聚合物聚n-异丙基丙烯酰胺(PIPAAm)组成的表面。为了推进该方法的临床应用,本研究试图确定一种最佳的人类ECM作为PIPAAm表面的涂层材料,它允许胰岛细胞附着在表面上,随后作为整体细胞片收获。将分散的大鼠胰岛细胞接种到涂有各种人层粘连蛋白同型的PIPAAm培养皿中:人层粘连蛋白(HL)-211、HL-332、HL-411、HL-511和HL-胎盘。第1天的镀效率,第3天的融合度,第3天的葡萄糖刺激胰岛素分泌试验。HL-332-PIPAAm组的电镀效率最高(83.1±0.7%)。HL-332-PIPAAm组细胞融合度最高(98.6±0.5%)。在HL-332-PIPAAm表面培养的胰岛细胞在葡萄糖刺激胰岛素分泌试验中表现出阳性反应。在HL-332-PIPAAm组中,通过将培养温度从37°C降低到20°C,细胞能够作为整体胰岛片收获。本研究表明,HL-332是PIPAAm涂层上制备胰岛细胞片的最佳人源ECM。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Human Laminin Isotype Coating for Creating Islet Cell Sheets.
Our experimental approach toward the development of new islet-based treatment for diabetes mellitus has been the creation of a monolayered islet cell construct (islet cell sheet), followed by its transplantation into a subcutaneous pocket. Previous studies describe rat laminin-5 (chain composition: α3, β3, γ2) as a suitable extracellular matrix (ECM) for surfaces comprised of a coated temperature-responsive polymer, poly(N-isopropylacrylamide) (PIPAAm). To progress toward the clinical application of this approach, the present study attempted to identify an optimal human ECM as a coating material on PIPAAm surfaces, which allowed islet cells to attach on the surfaces and subsequently to be harvested as a monolithic cell sheet. Dispersed rat islet cells were seeded onto PIPAAm dishes coated with various human laminin isotypes: human laminin (HL)-211, HL-332, HL-411, HL-511, and HL-placenta. Plating efficiency at day 1, the confluency at day 3, and glucose-stimulated insulin secretion test at day 3 were performed. The highest value of plating efficiency was found in the HL-332-PIPAAm group (83.1 ± 0.7%). The HL-332-PIPAAm group also showed the highest cellular confluency (98.6 ± 0.5%). Islet cells cultured on the HL-332-PIPAAm surfaces showed a positive response in the glucose-stimulated insulin secretion test. By reducing culture temperature from 37°C to 20°C in the HL-332-PIPAAm group, cells were able to be harvested as a monolithic islet sheet. The present study showed that HL-332 was an optimal human-derived ECM on a PIPAAm coating for preparing islet cell sheets.
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来源期刊
Cell medicine
Cell medicine MEDICINE, RESEARCH & EXPERIMENTAL-
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