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Culturing, passage, cryopreserving and resuscitating method for biopsy intestinal tumor organic and application thereof

A technology of intestinal tumors and culture methods, which is applied in the field of tumor organoid culture, can solve problems such as low survival rate and success rate, obstacles to wide application, and difficulty in establishing tumor transplantation models, achieving the effect of wide application and short test time

Inactive Publication Date: 2019-01-04
上海集技生物技术有限公司
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Problems solved by technology

However, we can see that the establishment of this tumor transplantation model is relatively difficult, especially for some tumors such as digestive tract tumors, because the original site is in the digestive tract cavity, the survival rate and success rate of transplantation are very low Therefore, it hinders its wide application and is only limited to some special tumors

Method used

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Experimental program
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Embodiment Construction

[0021] The principles and features of the present invention are described below, and the examples given are only used to explain the present invention, and are not intended to limit the scope of the present invention.

[0022] 1. Culture method of biopsied intestinal tumor organoids

[0023] 1. Place the biopsy tumor tissue of rice grain to soybean size in ice PBS (penicillin 100IU / ml+streptomycin 100μg / ml).

[0024] 2. Pre-cooled PBS (penicillin 100IU / ml+streptomycin 100μg / ml) for 5-10 minutes to wash several times, according to the cleanliness of the sample.

[0025] 3. Aseptically mince the tumor tissue, transfer it to the preheated enzyme digestion solution, and digest it in a water bath at 37°C for 30-60 minutes.

[0026] 4. After the digestion is completed, pipette the pipette repeatedly until the tumor fragments disappear, and centrifuge at 100g-300g for 5 minutes.

[0027] 5. Remove the supernatant and centrifuge repeatedly at 60gx for 5 minutes.

[0028] 6. Remove ...

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Abstract

The invention relates to a culturing, passage, cryopreserving and resuscitating method for the biopsy intestinal tumor organic and an application method thereof. Culturing and passage are conducted ona small amount of biopsy tumor tissue, an organoid with the genetic background highly consistent with that of source cancer tissue and the genetic mutation matched with the mutation level of a corresponding tissue sample can be cultured, the cultured organoid can be cryopreserved and resuscitated, the culturing and medicine effect testing time is shorter, compared with a PDTX model, the culturingcost is low, high throughput medicine screening can be achieved by building a tumor organoid sample library, the cultured organoid can test the ray sensibility of a sample and the like, and the application is wide.

Description

technical field [0001] The invention relates to a method for culturing tumor organoids, in particular to a method for culturing, passage, cryopreservation and recovery of biopsy intestinal tumor organoids and its application. Background technique [0002] In tumor research, especially in the screening of new drugs and the evaluation of individualized treatment, the PDTX model (human-derived tumor xenografting in the immune-deficient mouse model) has achieved certain results, and the transplanted tumor also retains the differentiation of the primary tumor The extent, morphological characteristics, structural characteristics, basic characteristics of molecular biology and tumor microenvironment. However, this model also has its shortcomings. Although the PDTX model simulates the biological characteristics of the primary tumor and can provide an in vivo simulation environment for tumor research, the establishment of this model is not completely the same as the primary tumor aft...

Claims

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Application Information

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IPC IPC(8): C12N5/09A01N1/02C12Q1/02
CPCA01N1/02G01N33/5005G01N33/5011G01N33/5044C12N5/0693C12N2513/00
Inventor 柏卫华祝远锋
Owner 上海集技生物技术有限公司
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