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  • Establishment and Long-Term Culture of Organoids Derived from Human Biliary Tract Carcinoma.

Establishment and Long-Term Culture of Organoids Derived from Human Biliary Tract Carcinoma.

STAR protocols (2020-10-29)
Yoshimasa Saito, Toshihide Muramatsu, Hidetsugu Saito
ABSTRACT

This protocol is a procedure for establishment and culture of cancer and non-cancer organoids using tissues from biliary tract carcinoma (BTC) patients. These BTC organoids can be used for various biological analyses and drug screening. One challenge in establishing and culturing BTC organoids is non-cancer cells contaminating surgically resected tumor tissues form organoids concurrently with cancer organoids. Careful validation that the established organoids are cancer-derived is important. For complete details on the use and generation of this protocol, please refer to Saito et al. (2019) in the journal Cell Reports.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Nicotinamide, BioReagent, suitable for cell culture, suitable for insect cell culture
Sigma-Aldrich
N-Acetyl-L-cysteine, suitable for cell culture, BioReagent
Sigma-Aldrich
[Leu15]-Gastrin I human, ≥95% (HPLC)
Sigma-Aldrich
Collagenase from Clostridium histolyticum, Type XI, 2-5 FALGPA units/mg solid, ≥800 CDU/mg solid
Sigma-Aldrich
R-Spondin1 expressing 293T Cell Line, The R-Spondin1-Expressing 293T cell line produces RSPO1, a critical protein used to establish 3D organoid cultures from stomach, small intestine, colon, pancreas, and liver sources.