Culture method of gynecological tumor primary cells

A technology for gynecological tumors and primary cells, which is applied in the field of culture of primary cells for gynecological tumors, and can solve the problems of long culture period, difficult removal of miscellaneous cells, and low success rate of culture

Pending Publication Date: 2020-09-04
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 gynecological tumor primary cells
  • Culture method of gynecological tumor primary cells
  • Culture method of gynecological tumor primary cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0082] Example 1. Preparation of reagents for culturing primary cells of gynecological tumors

[0083] 1. Sample preservation solution (100mL)

[0084] The specific formula of the sample preservation solution (100mL) is shown in Table 1.

[0085] Table 1 Sample Preservation Solution (100mL)

[0086]

[0087]

[0088] 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.

[0089] 2. Sample cleaning solution (100mL)

[0090] The specific formula of the sample cleaning solution (100mL) is shown in Table 2.

[0091] Table 2 Sample cleaning solution (100mL)

[0092]

[0093] The sample cleaning solution should be prepared and used immediately.

[0094] 3. Sample dissociation solution (10mL)

[0095] The specific formulation of the sample dissociation solution (10mL) is shown in Table 3.

[0096] Table 3 Sample Dissociation Solution (10mL)

[0097]

[0098]...

Embodiment 2

[0177] Example 2. Obtaining of postoperative specimens / biopsy puncture specimens / pleural and ascites samples of gynecological tumors

[0178] 1. Cooperate with tertiary hospitals, and the cooperation has passed the formal medical ethics review.

[0179] 2. The attending doctor 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 of the samples are: primary breast cancer, ovarian cancer, uterine Endometrial cancer, cervical cancer or its metastatic lesions, surgical specimens weighing more than 20mg, or pleural and ascites samples exceeding 100mL, or needle biopsy specimens exceeding 4 samples.

[0180] 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....

Embodiment 3

[0182] Example 3, Gynecological Tumor Solid Tumor Tissue Sample Dissociation Pretreatment

[0183] The following operations need to be performed on ice, and the entire operation steps need to be completed within 10 minutes.

[0184] The surgical equipment used in the following operations must be sterilized by high temperature and high pressure before use.

[0185] 1. Sample weighing.

[0186] 2. Clean the surface of the sample with 75% (volume percent) ethanol for 10 to 30 seconds.

[0187] 3. Wash the sample 5 times with sample cleaning solution, and wash the sample 5 times with sterile PBS solution.

[0188] 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 gynecological tumor primary cells. The invention provides a gynecological tumor primary cell culture method and a matched reagent. The core of the technology is as follows: a gynecological tumor solid tumor tissue is treated with a mild cell dissociation reagent, so that the activity of tumor cells in the tissue is ensured to the maximum extent; and a special serum-free culture medium is prepared, and solid tumor cells from a gynecological tumor are cultured in vitro by using a suspension culture system, so that interference of normal cells is eliminated to the maximum extent while normal amplification of the tumor cells is guaranteed. A gynecological tumor primary cell culture obtained by the method can be used for various cell-level in-vitro experiments, second-generation sequencing, animal model construction, cell line construction and the like. It is foreseeable that the culture method has wide application prospects in the fields of gynecological tumor research and clinical diagnosis treatment.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for culturing primary cells of gynecological tumors. Background technique [0002] Common gynecological tumors include breast cancer, ovarian cancer, and endometrial cancer. According to the statistics of the National Cancer Center in 2018, in 2014, breast cancer accounted for 16.5% of the incidence of female malignant tumors in my country, and the mortality rate reached 7.8%, ranking first and fifth among female tumors respectively. At present, the 5-year survival rate of breast cancer in my country is only 73.1% vs 90% (USA), which is still far behind developed countries. In addition, the incidence of ovarian cancer in my country accounts for 2.5% of female malignant tumors, and has increased by 30% in the past ten years. These gynecological tumors pose a severe challenge to the lives and health of women in my country. [0003] Although scientific research and medical i...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/09
CPCC12N5/0693C12N2501/11C12N2500/90C12N2501/115C12N2501/12C12N2501/392C12N2509/00C12N2501/91C12N2500/60
Inventor 尹申意张函槊
Owner SUZHOU GENOARRAY
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