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Lung cancer solid tumor primary cell culture method

A technology for primary cells and solid tumors, applied in cell culture active agents, tumor/cancer cells, culture process, etc., can solve the problems of difficult to remove miscellaneous cells, low culture success rate, and long culture cycle.

Inactive Publication Date: 2019-11-15
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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  • Lung cancer solid tumor primary cell culture method
  • Lung cancer solid tumor primary cell culture method
  • Lung cancer solid tumor primary cell culture method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0069] Embodiment 1, preparation is used for the reagent of culture lung cancer primary cell

[0070] 1. Sample preservation solution (100mL)

[0071] The specific formulation of the sample preservation solution (100mL) is shown in Table 1.

[0072] Table 1 Sample Preservation Solution (100mL)

[0073]

[0074] 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.

[0075] 2. Sample cleaning solution (100mL)

[0076] The specific formula of the sample cleaning solution (100mL) is shown in Table 2.

[0077] Table 2 Sample cleaning solution (100mL)

[0078]

[0079]

[0080] The sample cleaning solution should be prepared and used immediately.

[0081] 3. Sample dissociation solution (10mL)

[0082] The specific formulation of the sample dissociation solution (10mL) is shown in Table 3.

[0083] Table 3 Sample Dissociation Solution (10mL)

[0084]

[...

Embodiment 2

[0138] Embodiment 2, acquisition of postoperative specimen of lung cancer

[0139] 1. Cooperate with tertiary hospitals, and the cooperation has passed the formal medical ethics review.

[0140] 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 lung cancer, pathological stage II Or stage III, the pathological type is non-small cell lung cancer or small cell lung cancer, and the weight of the lung cancer sample exceeds 20mg.

[0141] 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's privacy are concealed and replaced with...

Embodiment 3

[0143] Embodiment 3, pre-dissociation treatment of lung cancer tissue samples

[0144] The following operations need to be performed on ice, and the entire operation steps need to be completed within 10 minutes.

[0145] The surgical equipment used in the following operations must be sterilized by high temperature and high pressure before use.

[0146] 1. Sample weighing.

[0147] 2. Clean the surface of the sample with 75% (volume percent) ethanol for 10 to 30 seconds.

[0148] 3. Wash the sample five times with sample cleaning solution, and wash the sample five times with sterile PBS solution.

[0149] 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 method for culturing lung cancer primary cells, and provides a lung cancer solid tumor primary cell culture method and a matched reagent. The technology is characterized by including the steps that (1) lung cancer solid tumor tissue is treated by using a mild cell dissociation reagent, and the activity of cancer cells in the tissue is ensured to the largest extent; (2) aspecial serum-free culture medium is prepared, in-vitro culture is conducted on the tumor cells derived from a lung cancer solid tumor source through a suspension culture system, and interference of normal cells is eliminated to the largest extent while normal amplification of the cancer cells is ensured. A lung cancer primary cell culture obtained through the method can be used for in-vitro experiments of various cell levels, secondary sequencing, construction of animal models, construction of cell lines and the like. It is contemplated that the culture method has wide application prospects in the fields of lung 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 lung cancer solid tumors. Background technique [0002] Lung cancer is the cancer with the highest morbidity and mortality in the world. The incidence and mortality of lung cancer in men are the highest among all malignant tumors, while the incidence and mortality of lung cancer in women are the second. At the same time, the incidence rate and death rate of lung cancer are the highest among all malignant tumors. It can be said that lung cancer is one of the most threatening malignant tumors to human health and life. [0003] Although scientific research and medical institutions around the world have invested heavily in research on the etiology and development of lung cancer, humans still know little about this disease. Lung cancer is a complex disease. Its occurrence and development are a dynamic process involving the interaction of many sign...

Claims

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Application Information

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IPC IPC(8): C12N5/09
CPCC12N5/0693C12N2500/25C12N2500/32C12N2500/34C12N2500/36C12N2500/38C12N2500/46C12N2501/11C12N2501/115C12N2501/392C12N2501/395C12N2501/71C12N2501/85C12N2501/998C12N2501/999
Inventor 席建忠尹申意李娟
Owner SUZHOU GENOARRAY
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