Chemotherapy-resistant Prostate Cancer Organoid Culture

CRPC with docetaxel chemotherapy-resistant patient-derived organoids was established pursuant to the method as described previously [51 (link),52 (link)]. The organoids were cultured with the advanced DMEM/F12 supplemented with 125 ng/ml rhR-spondin-1 (STEMCELL Technologies, 78213.1), 100 ng/ml rhNoggin (R&D Systems, 6057-NG-025), 1 ng/ml rhFGF (R&D Systems, 233-FB-025), rhFGF-10 (R&D Systems, 345-FG-025), 50 ng/ml (Meilunbio, China, MB8218-1), 1×N21 (R&D Systems, AR008), 10 μM Y-27632 (Abcam, Ab120129), 0.5 μM A83-01 (Beyotime Biotechnology, SF7917), 1:100 primocin (Invivogen, ant-pm-1), 10 μM SB202190 (Beyotime Biotechnology, SC0380), 10 mM Nicotinamide (Beyotime Biotechnology, S1761), and 1.25 mM N-acetylcysteine (Selleck, S1623), and the medium was changed every 2 to 3 days. Lentiviral transduction of AZGP1P2 shRNA and shRNA control was conducted according to the method as previously described [53 (link)]. The proliferation of organoids treated with 20 nM docetaxel was assessed by CCK-8 assay on the fifth day, and organoids were photographed using light microscopy. Three independent experiments were performed.

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Wang H., Liu J., Zhu X., Yang B., He Z, & Yao X. (2023). AZGP1P2/UBA1/RBM15 Cascade Mediates the Fate Determinations of Prostate Cancer Stem Cells and Promotes Therapeutic Effect of Docetaxel in Castration-Resistant Prostate Cancer via TPM1 m6A Modification. Research, 6, 0252.

Publication 2023

rhR-spondin-1 rhNoggin rhFGF rhFGF-10 MB8218-1 Y-27632 A83-01 ant-pm-1 SB202190 Nicotinamide

A83 01 Acetylcysteine Assay Cck 8 Chemotherapy Docetaxel Light microscopy Nicotinamide Organoids Patient Sb202190 Shrna Stemcell Y 27632

Corresponding Organization :

Other organizations : Tongji University, Shanghai Tenth People's Hospital, Inner Mongolia Medical University, Hunan Normal University, Shanghai Jiao Tong University

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Variable analysis

independent variables

Lentiviral transduction of AZGP1P2 shRNA and shRNA control

dependent variables

Proliferation of organoids treated with 20 nM docetaxel assessed by CCK-8 assay on the fifth day

control variables

CRPC with docetaxel chemotherapy-resistant patient-derived organoids established pursuant to the method as described previously
Organoids cultured with the advanced DMEM/F12 supplemented with 125 ng/ml rhR-spondin-1, 100 ng/ml rhNoggin, 1 ng/ml rhFGF, rhFGF-10, 50 ng/ml, 1×N21, 10 μM Y-27632, 0.5 μM A83-01, 1:100 primocin, 10 μM SB202190, 10 mM Nicotinamide, and 1.25 mM N-acetylcysteine, with medium changed every 2 to 3 days
Three independent experiments were performed

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