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

Active Publication Date: 2021-07-30
PEKING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] At present, there are few studies on kidney cancer organoids. The culture conditions of kidney cancer organoids are immature, and organoids of kidney cancer-related cells cannot be grown stably at the same time. Stable tumor embolism organoid characteristics can be achieved in vitro. The special role in the drug screening of kidney cancer organoids cannot stably detect the drug sensitivity of specific drugs. Therefore, in the field of basic medical research, there is a lack of an organoid screening model that can stably cultivate kidney cancer, paracancerous cells, and thrombus cells. In this way, drug sensitivity screening can be performed on kidney cancer-related cells at the same time, thereby enhancing the drug sensitivity and pertinence of clinical medication

Method used

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  • Organ-like combination related to kidney cancer and application thereof
  • Organ-like combination related to kidney cancer and application thereof
  • Organ-like combination related to kidney cancer and application thereof

Examples

Experimental program
Comparison scheme
Effect test

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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Abstract

The invention belongs to the field of medicine, and discloses a kidney cancer related organoid combination and application thereof. The kidney cancer related organoid combination comprises grouping establishment of kidney cancer related organoid, selection of screening drugs and establishment of a drug sensitivity endpoint, wherein the kidney cancer related organoid is from the body of a kidney cancer patient and is selected from perirenal cells, kidney cancer cells and kidney cancer embolus tissue; and then culture is carried out in a culture medium to obtain 2D and 3D models of three organs, and drug sensitivity screening is carried out to determine drug sensitivity characteristics of patients with different kidney cancer constitutions according to establishment of a drug sensitivity end point. The inventionprovides a screening model for determining medication preference for clinical medication.

Description

technical field [0001] The invention relates to the field of kidney cancer organoid culture, in particular to a combination of kidney cancer paracancerous tissue, kidney cancer tumor tissue and inferior vena cava tumor thrombus organoid, which belongs to the field of cell culture and drug screening. Background technique [0002] Renal cell carcinoma (RCC) is a common cancer originating in the renal epithelium, with an estimated 403,262 new cases and 175,098 deaths worldwide in 2018. RCC is a group of heterogeneous chemotherapy-resistant and radiation-resistant tumors, with more than 10 histological and molecular subtypes, among which clear cell carcinoma (clear cell RCC, ccRCC) is the most common, accounting for about 75%. A unique clinical manifestation of ccRCC is that it can grow into the renal vein or inferior vena cava and form a tumor thrombus (tumor thrombus, TT). Venous thrombosis is present in approximately 15% of ccRCC patients. The prognosis of TT patients is ve...

Claims

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Application Information

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IPC IPC(8): C12N5/09C12N5/071C12Q1/02G01N21/78
CPCC12N5/0697C12N5/0686C12N5/0693G01N33/5044G01N33/5011G01N21/78C12N2533/90C12N2513/00C12N2501/2302C12N2501/998C12N2501/11C12N2501/115C12N2500/32C12N2501/405C12N2503/02G01N2500/10
Inventor 孙晓娇雷艺芝刘振明张亮仁张树栋
Owner PEKING UNIV
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