Protocol Intestinal organoid co-culture protocol to study cell in vitro

被引：5
作者：
van Neerven, Sanne M. [1 ,2 ]
Ramadan, Rana [1 ,2 ]
Driel, Milou S. van [1 ,2 ]
Huels, David J. [1 ,2 ]
Vermeulen, Louis [1 ,2 ]
机构：
[1] Univ Amsterdam, Ctr Expt & Mol Med, Canc Ctr Amsterdam & Amsterdam Gastroenterol Endoc, Med Ctr,Lab Expt Oncol & Radiobiol, Meibergdreef 9, NL-1105 Amsterdam, Netherlands
[2] Oncode Inst, Meibergdreef 9, NL-1105 Amsterdam, Netherlands
来源：
STAR PROTOCOLS | 2022年 / 3卷 / 01期
基金：
欧洲研究理事会;
关键词：
Cancer; Cell Biology; Cell culture; Cell-based Assays; Flow Cytometry/Mass Cytometry; Microscopy; Organoids;
D O I：
10.1016/j.xpro.2021.101050
中图分类号：
Q5 [生物化学];
学科分类号：
071010 ; 081704 ;
摘要：
Intestinal organoid cultures are a powerful tool to study epithelial cells in vitro, as they are able to proliferate and differentiate into all cell lineages observed vivo. Co-culturing organoids with distinct genetic backgrounds provides an excellent approach to study contact dependent and independent interactions between healthy and mutant epithelial intestinal cells. Here, we provide 2D and 3D approaches to mouse organoid co-cultures using fluorescently labeled organoids and demonstrate the analysis of these co-cultures using flow cytometry and microscopy-based approaches. For complete details on the use and execution of this profile, please refer to van Neerven et al., 2021.
引用
收藏
页数：26
相关论文
共 11 条
  • [1] A Protocol for Lentiviral Transduction and Downstream Analysis of Intestinal Organoids
    de Jeude, Jooske F. Van Lidth
    Vermeulen, Jacqueline L. M.
    Montenegro-Miranda, Paula S.
    Van den Brink, Gijs R.
    Heijmans, Jarom
    [J]. JOVE-JOURNAL OF VISUALIZED EXPERIMENTS, 2015, (98):
  • [2] Efficient genetic engineering of human intestinal organoids using electroporation
    Fujii, Masayuki
    Matano, Mami
    Nanki, Kosaku
    Sato, Toshiro
    [J]. NATURE PROTOCOLS, 2015, 10 (10) : 1474 - 1485
  • [3] Active elimination of intestinal cells drives oncogenic growth in organoids
    Garcia, Ana Krotenberg
    Fumagalli, Arianna
    Le, Huy Quang
    Jackstadt, Rene
    Lannagan, Tamsin Rosemary Margaret
    Sansom, Owen J.
    van Rheenen, Jacco
    Suijkerbuijk, Saskia J. E.
    [J]. CELL REPORTS, 2021, 36 (01):
  • [4] Cytotoxicity and activation of the Wnt/beta-catenin pathway in mouse embryonic stem cells treated with four GSK3 inhibitors
    Naujok O.
    Lentes J.
    Diekmann U.
    Davenport C.
    Lenzen S.
    [J]. BMC Research Notes, 7 (1)
  • [5] Ramadan R, 2021, bioRxiv, DOI [10.1101/2021.04.14.439776, 10.1101/2021.04.14.439776, DOI 10.1101/2021.04.14.439776]
  • [6] Long-term Expansion of Epithelial Organoids From Human Colon, Adenoma, Adenocarcinoma, and Barrett's Epithelium
    Sato, Toshiro
    Stange, Daniel E.
    Ferrante, Marc
    Vries, Robert G. J.
    van Es, Johan H.
    van den Brink, Stieneke
    van Houdt, Winan J.
    Pronk, Apollo
    van Gorp, Joost
    Siersema, Peter D.
    Clevers, Hans
    [J]. GASTROENTEROLOGY, 2011, 141 (05) : 1762 - 1772
  • [7] Single Lgr5 stem cells build crypt-villus structures in vitro without a mesenchymal niche
    Sato, Toshiro
    Vries, Robert G.
    Snippert, Hugo J.
    van de Wetering, Marc
    Barker, Nick
    Stange, Daniel E.
    van Es, Johan H.
    Abo, Arie
    Kujala, Pekka
    Peters, Peter J.
    Clevers, Hans
    [J]. NATURE, 2009, 459 (7244) : 262 - U147
  • [8] Schwank G, 2016, METHODS MOL BIOL, V1422, P3, DOI 10.1007/978-1-4939-3603-8_1
  • [9] Apc-mutant cells act as supercompetitors in intestinal tumour initiation
    van Neerven, Sanne M.
    de Groot, Nina E.
    Nijman, Lisanne E.
    Scicluna, Brendon P.
    van Driel, Milou S.
    Lecca, Maria C.
    Warmerdam, Daniel O.
    Kakkar, Vaishali
    Moreno, Leandro F.
    Braga, Felipe A. Vieira
    Sanches, Delano R.
    Ramesh, Prashanthi
    ten Hoorn, Sanne
    Aelvoet, Arthur S.
    van Boxel, Marouska F.
    Koens, Lianne
    Krawczyk, Przemek M.
    Koster, Jan
    Dekker, Evelien
    Medema, Jan Paul
    Winton, Douglas J.
    Bijlsma, Maarten F.
    Morrissey, Edward
    Leveille, Nicolas
    Vermeulen, Louis
    [J]. NATURE, 2021, 594 (7863) : 436 - +
  • [10] Protocol for Application, Standardization and Validation of the Forskolin-Induced Swelling Assay in Cystic Fibrosis Human Colon Organoids
    Vonk, Annelotte M.
    van Mourik, Peter
    Ramalho, Anabela S.
    Silva, Iris A. L.
    Statia, Marvin
    Kruisselbrink, Evelien
    Suen, Sylvia W. F.
    Dekkers, Johanna F.
    Vleggaar, Frank P.
    Houwen, Roderick H. J.
    Mullenders, Jasper
    Boj, Sylvia F.
    Vries, Robert
    Amaral, Margarida D.
    Boeck, Kris de
    van der Ent, Cornelis K.
    Beekman, Jeffrey M.
    [J]. STAR PROTOCOLS, 2020, 1 (01):
12