Patient-derived organoids (MSK-PCa12) were generated and cultured as previously described (43 (link)). The tissues were deidentified prior to use. The 2 × 106 cells of MSK-Pca12 were engrafted subcutaneously into 6- to 8-wk-old CB17-SCID (severe combined immunodeficiency disease) male mice (Taconic). Mice were castrated when the tumor reached ∼200 mm3 and treated with ipatasertib (50 mg/kg) for 4 wk. Resistant tumors were collagenase/trypsin-digested into single cells and cultured in standard human organoid culture with ipatasertib (500 nM). AKT inhibitor ipatasertib was provided by Genentech under an MTA (material transfer agreement) and was dissolved in 0.5% methylcellulose and 0.2% Tween-80 for in vivo study. Acquisition of human tissue for prostate cancer organoids was performed under MSKCC IRB (Institutional Review Board of Memorial Sloan Kettering Cancer Center)-approved protocols 06-107 and 12-001.
Ipatasertib
Manufactured by Genentech
Sourced in China
Ipatasertib is a laboratory product used for research purposes. It is a small molecule inhibitor that targets the PI3K/AKT/mTOR signaling pathway. The core function of Ipatasertib is to inhibit the activity of the AKT kinase, which plays a crucial role in cell growth, proliferation, and survival.
Automatically generated - may contain errors
Lab products found in correlation
Paclitaxel, by Selleck Chemicals (2 mentions)
C b 17 scid, by Taconic Biosciences (1 mentions)
Taselisib, by Genentech (1 mentions)
Chloroquine diphosphate salt, by Merck Group (1 mentions)
Azd8186, by Selleck Chemicals (1 mentions)
Bkm120, by Selleck Chemicals (1 mentions)
Byl719, by LC Laboratories (1 mentions)
4 protocols using ipatasertib
1
Generation and Propagation of Patient-Derived Prostate Cancer Organoids
2
Dissolution and Storage of Chemotherapeutic Agents
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Paclitaxel was purchased from Selleck Chemicals (Selleckchem, Houston, TX, USA), Vinorelbine and eribulin from LookChem, Zhejiang J&C Biological Technology (China).
Taselisib and ipatasertib (GDC-0068) were provided by Genentech (Research proposal nr. OR-214726 for taselisib and nr. OR-214797 for ipatasertib).
They were dissolved in sterile dimethylsulfoxide (DMSO) and a 10 mM and 10 mM stock solutions were prepared and stored in aliquots at −20°C. Working concentrations were diluted in culture medium just before each experiment.
Taselisib and ipatasertib (GDC-0068) were provided by Genentech (Research proposal nr. OR-214726 for taselisib and nr. OR-214797 for ipatasertib).
They were dissolved in sterile dimethylsulfoxide (DMSO) and a 10 mM and 10 mM stock solutions were prepared and stored in aliquots at −20°C. Working concentrations were diluted in culture medium just before each experiment.
3
Cytotoxic Drug Preparation Protocol
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Paclitaxel was purchased from Selleck Chemicals (Selleckchem, Houston, TX, USA) and Chloroquine diphosphate salt from Sigma Aldrich. Taselisib (GDC0032) and ipatasertib (GDC-0068) were supplied by Genentech (Research proposal nr. OR-215703). They were dissolved in sterile dimethylsulfoxide (DMSO) and a 500 mM and 100 mM stock solutions, respectively, were prepared and stored in aliquots at −20 °C. Working concentrations were diluted in appropriate medium.
4
Inhibition of AR and PI3K Pathways
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The AR inhibitor MDV3100, enzalutamide, was synthesized by the MSKCC chemistry core. MDV3100 was used in our studies in vitro at a concentration of 10 mmol/L and in vivo with a dose of 10 mg/kg/day29 (link). The PI3K p110α-selective inhibitor BYL719 was purchased from LC Laboratories (cat# A-4477). BYL719 was dissolved in 0.5% methyl cellulose and dosed at 25 mg/kg daily P.O. The PI3K p110β-selective inhibitor AZD8186 was kindly provided by Laboratory of Neal Rosen and purchased from SelleckChem (cat# S7694). AZD8186 was dissolved in 0.5% HPMC + 0.1% Tween80 and dosed at 75 mg/kg b.i.d P.O. Pan-PI3K inhibitor BKM120 was purchased from SelleckChem (cat# S2247). BKM120 was dissolved in 0.5% methyl cellulose and dosed at 50 mg/kg daily P.O. IGF1R/INSR inhibitor linsitinib was purchased from LC Laboratory (cat# L-5814). Linsitinib was dissolved in 25 mM Tartaric Acid and dosed at 40 mg/kg daily P.O. AKT inhibitor ipatasertib was provided by Genentech under an MTA and was dissolved in 0.5% Methylcellulose and 0.2% Tween-80 for in vivo study.
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