Protocol for measuring transcytosis and recycling of IgG in intestinal epithelial Caco-2 cells and primary human intestinal organoids.
Protocol for measuring transcytosis and recycling of IgG in intestinal epithelial Caco-2 cells and primary human intestinal organoids.
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DOI：
10.1016/j.xpro.2023.102335
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发表时间：
2023-05-25
期刊：
影响因子：
--
通讯作者：
Lencer, Wayne I.
中科院分区：
文献类型：
--
作者：
Maeda, Keiko;Gwilt, Katlynn Bugda;Schmieder, Stefanie S.;Zachos, Nicholas C.;Lencer, Wayne I.
Transcytosis is the primary mechanism by which macro-molecules transverse epithelial cell barriers. Here, we present an assay for measuring transcytosis and recycling of IgG in intestinal epithelial Caco-2 cells and primary human intestinal organoids. We describe steps for establishing human enteroids or Caco-2 cells and plating monolayers. We then provide procedures for a transcytosis and recycling assay and a luciferase assay. The protocol facilitates quantification of membrane trafficking and can be used to probe endosomal compartments unique to polarized epithelia. For complete details on the use and execution of this protocol, please refer to Maeda K et al. (2022). Protocol for studying IgG trafficking across polarized human intestinal cells Quantification of transcytosis and recycling in intestinal epithelial cells Simple and sensitive assay can be applied to several types of human epithelial cells Allows for interrogation of epithelial cell functions and pathophysiological mechanisms Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. Transcytosis is the primary mechanism by which macro-molecules transverse epithelial cell barriers. Here, we present an assay for measuring transcytosis and recycling of IgG in intestinal epithelial Caco-2 cells and primary human intestinal organoids. We describe steps for establishing human enteroids or Caco-2 cells and plating monolayers. We then provide procedures for a transcytosis and recycling assay and a luciferase assay. The protocol facilitates quantification of membrane trafficking and can be used to probe endosomal compartments unique to polarized epithelia.
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DOI：
10.1083/jcb.201609035
发表时间：
2017-02
期刊：
The Journal of cell biology
影响因子：
--
作者：
Nelms B;Dalomba NF;Lencer W
通讯作者：
Lencer W
影响因子：
29.4
作者：
Sato, Toshiro;Stange, Daniel E.;Clevers, Hans
通讯作者：
Clevers, Hans
影响因子：
30.3
作者：
Maeda K;Zachos NC;Orzalli MH;Schmieder SS;Chang D;Bugda Gwilt K;Doucet M;Baetz NW;Lee S;Crawford SE;Estes MK;Kagan JC;Turner JR;Lencer WI
通讯作者：
Lencer WI
影响因子：
--
作者：
Staab JF;Lemme-Dumit JM;Latanich R;Pasetti MF;Zachos NC
通讯作者：
Zachos NC
影响因子：
64.8
作者：
Sato, Toshiro;Vries, Robert G.;Clevers, Hans
通讯作者：
Clevers, Hans