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

RESUMEN

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.

MATERIALES

Referencia del producto

Marca

Descripción del producto

A9165

Sigma-Aldrich

N-Acetyl-L-cysteine, suitable for cell culture, BioReagent

N0636

Sigma-Aldrich

Nicotinamide, BioReagent, suitable for cell culture, suitable for insect cell culture

C7657

Sigma-Aldrich

Colagenasa from Clostridium histolyticum, Type XI, 2-5 FALGPA units/mg solid, ≥800 CDU/mg solid

G9145

Sigma-Aldrich

[Leu¹⁵]-Gastrin I human, ≥95% (HPLC)

SCC111

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.