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Culture medium for culturing recurrent renal cell carcinoma organoid and application of culture medium

A technology of culture medium and kidney cells, applied in the direction of urinary tract/kidney cells, culture process, tissue culture, etc., can solve the problems of increasing difficulty in recurrent kidney cancer organoid culture, and achieve the effect of reducing the cost of culture medium and reducing dependence

Inactive Publication Date: 2022-06-03
上海诺典生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Compared with the culture of untreated primary kidney cancer organoids, the difficulty of culturing recurrent kidney cancer organoids will increase theoretically, and there is no related research on the culture of recurrent kidney cancer organoids

Method used

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  • Culture medium for culturing recurrent renal cell carcinoma organoid and application of culture medium
  • Culture medium for culturing recurrent renal cell carcinoma organoid and application of culture medium

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] Example 1. Improved culture medium 1

[0058] F12K medium: 1640 medium (4:1), L-Glutathine (50mg / L), sodium selenite (0.1mg / L), ethanolamine hydrochloride (100mg / L), insulin (5mg / L), β-Mercaptoethanol (30 mM);

[0059] R-spondin 1 (400ng / ml), Noggin (100ng / ml), N2:B27 (1:2), GlutaMAXSupplement (1×), HEPES (1×), Nicotinamide (15mM), Normocin TM (1×), N-acetyl cysteine ​​(1 mM);

[0060] A83-01 (2 μM), EGF (100 ng / ml), DMSO (0.5%), BMP2 (50 ng / ml), FGF10 (150 ng / ml).

Embodiment 2

[0061] Example 2. Improved culture medium 2

[0062] F12K medium: 1640 medium (2:1), L-Glutathine (50mg / L), sodium selenite (0.2mg / L), ethanolamine hydrochloride (150mg / L), insulin (15mg / L), β-Mercaptoethanol (40mM);

[0063] R-spondin 1 (250ng / ml), Noggin (50ng / ml), N2:B27 (2:1), GlutaMAX Supplement (1x), HEPES (1x), Nicotinamide (20mM), Normocin TM (1×), N-acetylcysteine ​​(5mM);

[0064] EGF (50ng / ml), FGF9 (100ng / ml), BMP4 (250ng / ml), BMP7 (150ng / ml), heparin (20IU / ml).

Embodiment 3

[0065] Example 3. Comparison of Organoid Culture and Drug Sensitivity Screening for Recurrent Renal Cell Carcinoma Tumor Samples Cultured in Two Mediums of the Present Invention and Primary Renal Cell Carcinoma Medium.

[0066] Different recurrent renal cell carcinoma tumor samples were cultured in the medium described in the above Examples 1 and 2 and the traditional primary renal cell carcinoma medium (Nature Communications, NC), and the organoid success rate and organoid morphology were observed.

[0067] The specific operation method is as follows:

[0068] 1. The puncture samples of 12 cases of recurrent renal cell carcinoma were stored in a special preservation solution at 4°C, and transferred to the operating room at low temperature within 24 hours;

[0069] 2. The samples were washed 3-5 times with PBS on ice, 5min each time;

[0070] 3. The samples were cut into 1mm fragments at low temperature and placed in pre-warmed collagenase digestion solution at 37°C, and the ...

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Abstract

The invention discloses a culture medium for a recurrent renal cell carcinoma tumor organoid and a culture method of the culture medium. By adjusting the content of each component of the culture medium, the organoid culture of a tumor sample of the recurrent renal cell carcinoma can be realized, the success rate of the culture of the organoid of the renal cell carcinoma is greatly improved, the culture period is shortened, the culture cost is reduced, and a foundation is laid for revealing the pathogenesis of the recurrent renal cell carcinoma and screening anti-cancer drugs.

Description

technical field [0001] The invention relates to the field of cell culture, in particular to a culture medium for recurrent renal cell carcinoma organoids and a culture method thereof. Background technique [0002] Kidney cancer is one of the more common malignant tumors. According to the global cancer statistics in 2020, the annual number of new cases of kidney cancer is more than 431,000, and the annual number of deaths is about 180,000. The incidence of kidney cancer is second only to prostate cancer and bladder cancer, ranking third in urinary system tumors. Renal cancer is traditionally divided into three pathological types: clear cell carcinoma, papillary renal cell carcinoma, and chromophobe renal cell carcinoma. The most common pathological type of renal cancer is clear cell carcinoma of the kidney, accounting for about 80-90% of the total number of renal cancers. The current treatment options for kidney cancer are a combination of surgery, radiotherapy, chemothera...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/09C12N5/071
CPCC12N5/0686C12N5/0693C12N2501/998C12N2500/12C12N2500/30C12N2500/44C12N2501/33C12N2501/415C12N2501/155C12N2500/32C12N2500/60C12N2500/46C12N2501/999C12N2501/727C12N2501/11C12N2500/62C12N2501/119C12N2501/91C12N2509/00C12N2503/02
Inventor 柏卫华
Owner 上海诺典生物科技有限公司
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