胰腺导管类器官的生成及完整类器官的全载免疫染色

Q3 Biochemistry, Genetics and Molecular Biology Current Protocols in Cell Biology Pub Date : 2018-12-12 DOI:10.1002/cpcb.82
Habib Rezanejad, Jennifer Hollister Lock, Brooke A Sullivan, Susan Bonner-Weir
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引用次数: 7

摘要

传统上,细胞和组织的研究是通过在二维培养皿或烧瓶中培养分离的原代细胞或永生化细胞系来进行的。然而,关于2D培养的一个警告是,细胞很难再现其体内对应物,这主要是由于缺乏3D细胞-细胞和细胞-细胞外基质的相互作用。近年来,体外类器官作为三维培养物作为研究不同组织的模型得到了广泛关注。在成人中,胰腺导管细胞被认为是干细胞或祖细胞的来源,因此开发新的方法来研究导管细胞将是有用的。在这里,我们提供了一种分离小鼠胰腺导管细胞的方案,以及一种从这些导管细胞中产生三维类器官结构的经济有效的方案。此外,我们还设计了一种无需切片的完整类器官免疫染色方案。©2018 by John Wiley &儿子,Inc。
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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Generation of Pancreatic Ductal Organoids and Whole-Mount Immunostaining of Intact Organoids

Traditionally, studies of cells and tissues have been performed on isolated primary cells or immortalized cell lines by culturing them in 2D culture dishes or flasks. However, a caveat regarding 2D culture is that the cells poorly recapitulate their in vivo counterparts, mainly due to a lack of 3D cell-cell and cell–extracellular matrix interactions. In recent years, the development of in vitro organoids as 3D culture has gained substantial attention as a model to study different tissues. In adults, pancreatic ductal cells are considered as a source of stem or progenitor cells, so developing new methods to study ductal cells would be useful. Here, we provide a protocol to isolate mouse pancreatic ductal cells and a cost-effective protocol to generate 3D organoid structures from such ductal cells. Additionally, we have devised a protocol for immunostaining of intact whole organoids without sectioning. © 2018 by John Wiley & Sons, Inc.

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Current Protocols in Cell Biology
Current Protocols in Cell Biology Biochemistry, Genetics and Molecular Biology-Cell Biology
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期刊介绍: Developed by leading scientists in the field, Current Protocols in Cell Biology is an essential reference for researchers who study the relationship between specific molecules and genes and their location, function and structure at the cellular level. Updated every three months in all formats, CPCB is constantly evolving to keep pace with the very latest discoveries and developments.
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