Organ-like combination related to kidney cancer and application thereof
A technology of organoids and kidney cancer, applied in the fields of kidney cancer organoid culture, cell culture and drug screening, can solve the problems of lack, enhance the sensitivity and pertinence of clinical drugs, immature culture conditions of kidney cancer organoids, etc., to achieve The effect of reducing tissue volume requirements
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Embodiment 1
[0054] Culture process of a kidney cancer organoid combination
[0055] 1. Wash tumor biopsy tissue samples twice with PBS; patients with clinical renal cancer and inferior vena cava thrombosis
[0056] 2. Remove necrotic mucus tissue;
[0057] 3. The remainder was minced with sterile scissors in ADDF+++ (advanced DMEM / F12 containing 1× Glutamax, 10 mM HEPES, and antibiotics) solution containing 1 mg / mL collagenase (Sigma, C9407) and 10 μM Y-27632. For small pieces of about 5mm, incubate at 37°C for 1 hour, and pipette the lysate every 15 minutes to promote digestion;
[0058] Note: ADDF+++ medium: Advanced DMEM / F12+1×Glutamax+10mM HEPES+antibiotics+1mg / mL collagenase (Sigma, C9407)+10μM Y-27632. (The antibiotic is a combination of penicillin + streptomycin, the concentration is: the working concentration of penicillin is 100U / mL, and the working concentration of streptomycin is 0.1mg / mL)
[0059] In all subsequent steps, the pipette tip must be pre-wetted with DMEM+1%BSA b...
Embodiment 2
[0075] Using 10 patients with kidney cancer, the success rate (9 / 10): 10 patients, 1 patient failed to culture the tumor thrombus of kidney cancer, and cultured kidney cancer tissue and kidney tissue organoids successfully;
[0076] Difficulty of maintenance: cultured in a 37°C, 5% CO2 incubator, and the kidney organoid growth medium is updated every 3-4 days as needed, with a high success rate.
Embodiment 3
[0078] The specific screening drugs are as follows in Table 1
[0079] Table 1 is the screened drugs and their data
[0080]
[0081]
[0082] Dose-response curve: The concentration of candidate drugs in the drug screening based on the organoid model in this study is determined through the dose-effect curve relationship, based on the drug concentration as the abscissa, and the effect strength as the ordinate to judge the dose of the drug. Candidate evaluation drugs are all Select the amount of drug within the safe range of treatment for drug susceptibility testing.
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