Three-dimensional culture of mouse pancreatic islet on a liver-derived perfusion-decellularized bioscaffold for potential clinical application
Three-dimensional culture of mouse pancreatic islet on a liver-derived perfusion-decellularized bioscaffold for potential clinical application
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肝源性灌注脱细胞生物支架上小鼠胰岛的三维培养及其潜在的临床应用
DOI：
10.1177/0885328215587610
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发表时间：
2015-05
影响因子：
2.9
通讯作者：
Wang ZW
中科院分区：
文献类型：
--
作者：
Xu Tianxin;Zhu Mingyan;Wu Di;Huang Yan;Fan Xiangjun;Zhu Shajun;Lin Changchun;Zhou Pengcheng;Lu Yuhua;Wang Zhiwei;Guo Yibing;Lin Changchun;Li Xiaohong;Lu Jingjing;Zhu Hui;Lu Yuhua;Wang Zhiwei;Wang ZW
三维肝脏脱细胞生物支架的前沿技术有可能提供适合驻留细胞的微环境，甚至开发新的功能器官。肝脏脱细胞生物支架保留了天然细胞外基质和三维结构，以支持细胞培养。本研究的目的是发现来自小鼠肝脏的三维细胞外基质是否可以促进小鼠离体胰岛的生长和维持生理功能。我们使用1%Triton X-100/0.1%铵方案制备了一种全器官肝脏脱细胞生物支架，其可以成功地保留细胞外基质蛋白和天然血管通道。为了评价脱细胞肝脏作为胰岛移植支架的潜力，将脱细胞肝脏生物支架与通过胶原酶P消化方案获得的小鼠原代胰岛输注。通过显微镜分析、双硫腙染色、胰岛素免疫荧光和葡萄糖刺激实验来评估其产量、形态和质量。比较肝脏脱细胞生物支架三维培养与传统二维平板培养的差异，并进行苏木精-伊红染色、免疫组化和胰岛素基因表达检测。我们的结果表明，肝脏脱细胞生物支架可以支持细胞培养和维持细胞功能。与传统的二维培养体系相比，三维培养体系可使胰岛素基因表达上调。这些结果表明，通过灌注脱细胞技术的肝脏生物支架可以作为一个平台，以支持胰岛在体外的生存和功能。同时三维培养体系相对于二维培养具有上级作用。这项研究推进了再生医学领域对肝脏脱细胞生物支架的发展，能够形成一个新的器官，并可用于潜在的临床应用。
The cutting-edge technology of three-dimensional liver decellularized bioscaffold has a potential to provide a microenvironment that is suitable for the resident cells and even develop a new functional organ. Liver decellularized bioscaffold preserved the native extracellular matrix and three-dimensional architecture in support of the cell culture. The goal of this study was to discover if three-dimensional extracellular matrix derived from mouse liver could facilitate the growth and maintenance of physiological functions of mouse isolated islets. We generated a whole organ liver decellularized bioscaffold which could successfully preserve extracellular matrix proteins and the native vascular channels using 1% Triton X-100/0.1% ammonium protocol. To evaluate the potential of decellularized liver as a scaffold for islets transplantation, the liver decellularized bioscaffold was infused with mouse primary pancreatic islets which were obtained through Collagenase P digestion protocol. Its yield, morphology, and quality were estimated by microscopic analysis, dithizone staining, insulin immunofluorescence and glucose stimulation experiments. Comparing the three-dimensional culture in liver decellularized bioscaffold with the orthodoxy two-dimensional plate culture, hematoxylin-eosin staining, immunohistochemistry, and insulin gene expression were tested. Our results demonstrated that the liver decellularized bioscaffold could support cellular culture and maintenance of cell functions. In contrast with the conventional two-dimensional culture, three-dimensional culture system could give rise to an up-regulated insulin gene expression. These findings demonstrated that the liver bioscaffold by a perfusion-decellularized technique could serve as a platform to support the survival and function of the pancreatic islets in vitro. Meanwhile three-dimensional culture system had a superior role in contrast with the two-dimensional culture. This study advanced the field of regenerative medicine towards the development of a liver decellularized bioscaffold capable of forming a neo-organ and could be used as potential clinical application.
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