Protocol for measuring transcytosis and recycling of IgG in intestinal epithelial Caco-2 cells and primary human intestinal organoids

被引：1

作者：

Maeda, Keiko ^{[1
,4
]}

Gwilt, Katlynn Bugda ^{[1
]}

Schmieder, Stefanie S. ^{[1
]}

Zachos, Nicholas C. ^{[2
]}

Lencer, Wayne I. ^{[1
,3
]}

机构：

[1] Boston Childrens Hosp, Div Gastroenterol, Boston, MA 02115 USA

[2] Johns Hopkins Univ, Dept Med, Div Gastroenterol & Hepatol, Sch Med, Baltimore, MD 21205 USA

[3] Harvard Med Sch, Harvard Digest Dis Ctr, Boston, MA 02115 USA

[4] Nagoya Univ, Dept Endoscopy, Grad Sch Med, 65 Tsurumai Cho,Showa Ku, Nagoya 4668550, Japan

来源：

STAR PROTOCOLS | 2023年 / 4卷 / 02期

关键词：

Cell Biology; Molecular Biology;

D O I：

10.1016/j.xpro.2023.102335

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

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).1

引用

页数：16