Biotechnology and Drug Development Research Laboratory, School of Pharmacy, Curtin Health Innovation Research Institute (CHIRI), Curtin University, Perth, WA, Australia.
School of Public Health, Curtin Health Innovation Research Institute, Curtin University, Perth, WA, Australia.
Pharm Res. 2017 Jun;34(6):1217-1223. doi: 10.1007/s11095-017-2138-y. Epub 2017 Mar 13.
Recently we demonstrated that microencapsulation of a murine pancreatic β-cell line using an alginate-ursodeoxycholic acid (UDCA) matrix produced microcapsules with good stability and cell viability. In this study, we investigated if translation of this formulation to microencapsulation of primary β-cells harvested from mature double-transgenic healthy mice would also generate stable microcapsules with good cell viability.
Islets of Langerhans were isolated from Ngn3-GFP/RIP-DsRED mice by intraductal collagenase P digestion and density gradient centrifugation, dissociated into single cells and the β-cell population purified by Fluorescence Activated Cell Sorting. β-cells were microencapsulated using either alginate-poly-l-ornithine (F1; control) or alginate-poly-l-ornithine-UDCA (F2; test) formulations. Microcapsules were microscopically examined and microencapsulated cells were analyzed for viability, insulin and cytokine release, 2 days post-microencapsulation.
Microcapsules showed good uniformity and morphological characteristics and even cell distribution within microcapsules with or without UDCA. Two days post microencapsulation cell viability, mitochondrial ATP and insulin production were shown to be optimized in the presence of UDCA whilst production of the proinflammatory cytokine IL-1β was reduced. Contradictory to our previous studies, UDCA did not reduce production of any other pro-inflammatory biomarkers.
These results suggest that UDCA incorporation improves microcapsules' physical and morphological characteristics and improves the viability and function of encapsulated mature primary pancreatic β-cells.
最近我们证明，使用藻酸盐-熊去氧胆酸（UDCA）基质对小鼠胰岛β细胞系进行微囊化可产生具有良好稳定性和细胞活力的微胶囊。在这项研究中，我们研究了从成熟的双转基因健康小鼠中收获的原代β细胞的这种制剂的翻译是否也会产生具有良好细胞活力的稳定微胶囊。
通过胰管内胶原酶 P 消化和密度梯度离心从 Ngn3-GFP/RIP-DsRED 小鼠中分离胰岛，将胰岛解离成单细胞，并通过荧光激活细胞分选纯化β细胞。使用藻酸盐-聚-L-鸟氨酸（F1；对照）或藻酸盐-聚-L-鸟氨酸-UDCA（F2；测试）制剂对β细胞进行微囊化。对微胶囊进行显微镜检查，并在微囊化后 2 天分析微囊化细胞的活力、胰岛素和细胞因子释放。
微胶囊显示出良好的均匀性和形态特征，并且即使在有或没有 UDCA 的情况下，微胶囊内的细胞分布也很均匀。微囊化后 2 天，细胞活力、线粒体 ATP 和胰岛素的产生在 UDCA 的存在下得到优化，而促炎细胞因子 IL-1β 的产生减少。与我们之前的研究相反，UDCA 并没有减少任何其他促炎生物标志物的产生。
这些结果表明，UDCA 的加入改善了微胶囊的物理和形态特征，并提高了封装成熟原代胰腺β细胞的活力和功能。
