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A kidney cancer-related organoid combination and its application

A technology of organoids and kidney cancer, applied in cell culture and drug screening, and the field of kidney cancer organoid culture, can solve the problems of shortage, immature culture conditions of kidney cancer organoids, enhance the sensitivity and pertinence of clinical drugs, and achieve The effect of reducing tissue volume requirements

Active Publication Date: 2022-06-28
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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  • A kidney cancer-related organoid combination and its application
  • A kidney cancer-related organoid combination and its application
  • A kidney cancer-related organoid combination and its application

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; clinical renal cancer patients with inferior vena cava thrombosis

[0056] 2. Removal of necrotic mucous tissue;

[0057] 3. The remainder was minced using sterile scissors in ADDF++ (advanced DMEM / F12 with 1×Glutamax, 10mM HEPES and antibiotics) solution containing 1 mg / mL collagenase (Sigma, C9407) and 10 μM Y-27632. Small pieces of about 5mm, incubate at 37°C for 1 hour, and aspirate the lysate every 15 minutes to facilitate digestion;

[0058] Note: ADDF+++ medium: Advanced DMEM / F12+1×Glutamax+10mM HEPES+antibiotics+1 mg / mL collagenase (Sigma, C9407)+10 μM Y-27632. (The antibiotic is a combination of penicillin + streptomycin, and the concentration is: the working concentration of penicillin is 100U / mL, and the working concentration of streptomycin is 0.1 mg / mL)

[0059] For all subsequent steps, the pipette tip needs to be pre-wetted with DMEM+1% ...

Embodiment 2

[0075] Using 10 patients with renal cancer, the success rate (9 / 10): 10 patients were unsuccessful in the culture of renal cancer tumor thrombus, and both renal cancer tissue and renal tissue organoids were successfully cultured;

[0076] Maintenance Difficulty: Culture in a 37°C, 5% CO2 incubator. As needed, the kidney organoid growth medium is renewed every 3-4 days, with a high success rate.

Embodiment 3

[0078] The specific screening drugs are as follows in Table 1

[0079] Table 1 shows 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 was determined by the relationship of the dose-response curve. Based on the drug concentration as the abscissa and the action intensity as the ordinate, the drug dose was judged, and the candidate evaluation drugs were all evaluated. Select the amount of drug within the therapeutically safe range for drug susceptibility testing.

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Abstract

The invention belongs to the field of medicine, and discloses a combination of kidney cancer-related organoids and its application, including the establishment of grouping of kidney cancer-related organoids, the selection of screening drugs, and the establishment of drug sensitivity endpoints, wherein the kidney cancer-related organoids From renal cancer patients, they were selected from pararenal cells, renal cancer cells, and tumor embolus tissues of renal cancer, and were cultured in culture medium to obtain three organoid 2D and 3D models for drug sensitivity screening. The establishment of the end point can determine the drug sensitivity characteristics of patients with different kidney cancer constitutions, and provide a screening model for determining drug preference for clinical medication.

Description

technical field [0001] The invention relates to the field of renal carcinoma organoid culture, in particular to a combination comprising renal carcinoma adjacent tissue, renal carcinoma tumor tissue and inferior vena cava tumor thrombus organoid, belonging to the fields of cell culture and drug screening. Background technique [0002] Renal cell carcinoma (RCC) is a common cancer originating from the renal epithelium, with an estimated 403,262 new cases and 175,098 deaths worldwide in 2018. RCC is a heterogeneous group of chemotherapy-resistant and radio-resistant tumors with more than 10 histological and molecular subtypes, among which 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 tumor thrombus (TT). Venous thrombosis is present in approximately 15% of ccRCC patients. The prognosis for patients with TT is poor, with a median survival of o...

Claims

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Application Information

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Patent Type & Authority Patents(China)
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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