Gallbladder Cancer Organoid Culture

GBC-PDOs cultures were established from tumor tissue samples of patients diagnosed with moderated differentiated and pT3 N0/N1 Mx stage (Eight edition of American Joint Committee on Cancer (AJCC)) advanced tubular GBC. Gallbladder tumor organoids were generated as previously described [54 (link)]. For organoids cultures, dissociated cells were washed in 1× DPBS and embedded in growth factor reduced Matrigel (#356231, Corning, NY, USA) and cultured in complete media (Intesticult (#6005, Stemcell Technologies, Vancouver, BC, Canada), 0.5 μmol/L A83-01 (#SML0788; Sigma-Aldrich), 100 ng/mL, FGF10 (#100-26, PeproTech, Inc., Rocky Hill, NJ, USA), 100 ng/mL HGF (#100-39; Peprotech), 10 nmol/L Gastrin I (#G9145; Sigma), 10 mmol/L N-acetyl-l-cysteine (#A9165; Sigma-Aldrich) 10 mmol/L nicotinamide (#N0636; Sigma-Aldrich), 1× B27 supplement (#17504-044; Gibco, Thermo Fisher Scientific, Waltham, MA, USA), 1× N2 supplement (#175020-048; Gibco), 1 mg/mL Primocin (#ant-pm-1; InvivoGen, San Diego, CA, USA) and 10.5 μmol/L Y-27632 (#Y0503, Sigma-Aldrich). Organoids were passaged via mechanical dissociation and passage was performed weekly with a 1:3 ratio.

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García P., Rosa L., Vargas S., Weber H., Espinoza J.A., Suárez F., Romero-Calvo I., Elgueta N., Rivera V., Nervi B., Obreque J., Leal P., Viñuela E., Aguayo G., Muñiz S., Sagredo A., Roa J.C, & Bizama C. (2020). Hippo-YAP1 Is a Prognosis Marker and Potentially Targetable Pathway in Advanced Gallbladder Cancer. Cancers, 12(4), 778.

Publication 2020

Matrigel Intesticult A83-01 FGF10 HGF Gastrin I N-acetyl-l-cysteine nicotinamide B27 supplement N2 supplement Primocin Y-27632

A83 01 Cancer Cells Cultured media Fgf10 Gallbladder tumor Gastrin i Growth factor Joint L cysteine Matrigel Nicotinamide Organoids Patients Stemcell Supplement Tissue Tumor Y 27632

Corresponding Organization :

Other organizations : Pontificia Universidad Católica de Chile, Universidad de La Frontera, Science for Life Laboratory, Karolinska Institutet, University of Illinois at Chicago, Complejo Asistencial Sótero del Río, Millennium Institute on Immunology and Immunotherapy

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Variable analysis

independent variables

Dissociated cells were embedded in growth factor reduced Matrigel
Cells were cultured in complete media containing Intesticult, A83-01, FGF10, HGF, Gastrin I, N-acetyl-l-cysteine, nicotinamide, B27 supplement, N2 supplement, Primocin, and Y-27632

dependent variables

Generation of gallbladder tumor organoids

control variables

Passage of organoids was performed weekly with a 1:3 ratio

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