Culture method of colorectal cancer solid tumor primary cells

A colorectal cancer and primary cell technology, applied in the biological field, can solve the problems of long culture cycle, difficult to remove miscellaneous cells, and low culture success rate

Inactive Publication Date: 2019-12-20
SUZHOU GENOARRAY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Existing primary tumor cell culture technologies mainly include 2D culture, 3D culture, reprogramming culture, etc. These methods face the problems of extremely long culture cycle, low culture success rate, and difficulty in removing stray cells to varying degrees.

Method used

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  • Culture method of colorectal cancer solid tumor primary cells
  • Culture method of colorectal cancer solid tumor primary cells
  • Culture method of colorectal cancer solid tumor primary cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0073] Example 1. Preparation of reagents for culturing primary colorectal cancer cells

[0074] 1. Sample preservation solution (100mL)

[0075] The specific formulation of the sample preservation solution (100mL) is shown in Table 1.

[0076] Table 1 Sample Preservation Solution (100mL)

[0077]

[0078] After the sample preservation solution is prepared, it is divided into 15mL centrifuge tubes, 5mL per tube. After aliquoting, it can be stored at 4°C for 1 month.

[0079] 2. Sample cleaning solution (100mL)

[0080] The specific formula of the sample cleaning solution (100mL) is shown in Table 2.

[0081] Table 2 Sample cleaning solution (100mL)

[0082]

[0083] The sample cleaning solution should be prepared and used immediately.

[0084] 3. Sample dissociation solution (10mL)

[0085] The specific formulation of the sample dissociation solution (10mL) is shown in Table 3.

[0086] Table 3 Sample Dissociation Solution (10mL)

[0087]

[0088] Note: The s...

Embodiment 2

[0163] Example 2. Obtaining of postoperative specimens of colorectal cancer

[0164] 1. Cooperate with tertiary hospitals, and the cooperation has passed the formal medical ethics review.

[0165] 2. The attending physician selects the patients according to the clinical indications stipulated in the medical guidelines, and selects appropriate samples for in vitro culture according to the clinical indications during the operation. The selection criteria for the samples are: primary colorectal cancer, pathological stage: Stage II, III or IV, various pathological types of colorectal cancer or metastatic lesions of colorectal cancer, and colorectal cancer surgical specimens weighing more than 20 mg.

[0166] 3. The attending doctor provides basic clinical information such as the patient's gender, age, medical history, family history, smoking history, pathological staging and typing, and clinical diagnosis. The patient's name, ID number and other information related to the patient...

Embodiment 3

[0168] Embodiment 3, pre-dissociation treatment of colorectal cancer tissue samples

[0169] The following operations need to be performed on ice, and the entire operation steps need to be completed within 10 minutes.

[0170] The surgical equipment used in the following operations must be sterilized by high temperature and high pressure before use.

[0171] 1. Sample weighing.

[0172] 2. Clean the surface of the sample with 75% (volume percent) ethanol for 10 to 30 seconds.

[0173] 3. Wash the sample 10 times with sample cleaning solution and 5 times with sterile PBS solution.

[0174] 4. Use ophthalmic scissors, ophthalmic forceps, scalpel and other equipment to carefully peel off the adipose tissue, connective tissue, and necrotic tissue in the sample.

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Abstract

The invention discloses a culture method of colorectal cancer primary cells. The invention provides the culture method of colorectal cancer primary cells and an auxiliary reagent. The core of the method comprises the following steps: (1) treating colorectal cancer solid tumor tissues by using a mild cell dissociation reagent to ensure the vitality of cancer cells in the tissues to the greatest extent; and (2) preparing a special serum-free culture medium, and carrying out in-vitro culture on tumor cells derived from colorectal cancer solid tumors by utilizing a suspension culture system, so that the interference of normal cells is eliminated to the greatest extent while the normal amplification of cancer cells is ensured. The colorectal cancer primary cell culture obtained by the method can be used for in-vitro experiments of various cell levels, next-generation sequencing, animal model construction, cell line construction and the like. Predictably, the culture method has wide application prospects in the fields of colorectal cancer research and clinical diagnosis and treatment.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for culturing primary cells of colorectal cancer solid tumors. Background technique [0002] Colorectal cancer is one of the most common malignant tumors that seriously threaten human health. In my country, the incidence rate of colorectal cancer is 9.24%, ranking fourth among all malignant tumors. The mortality rate of colorectal cancer is 11.77%, ranking fifth among all malignant tumors. With the development of the economy, the improvement of living standards and the change of lifestyle, the incidence of colorectal cancer will continue to rise. In addition, the risk of recurrence and metastasis of colorectal cancer is high, and more than 50% of colorectal cancer patients will experience recurrence and metastasis of varying degrees within months to years after radical treatment. [0003] Although scientific research and medical institutions around the world have invested...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/09
CPCC12N5/0693C12N2509/00
Inventor 张函槊尹申意
Owner SUZHOU GENOARRAY
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