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Culture method of gastric cancer micro-tumor cell model

A gastric cancer and tumor technology, applied in the field of culturing gastric cancer microtumor cell models, can solve the problems of inability to reproduce the tumor microenvironment, difficult to remove miscellaneous cells, low culture success rate, etc. High stability effect

Pending Publication Date: 2021-12-03
SUZHOU GENOARRAY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Therefore, it is very important to develop a tumor model that can accurately reflect the characteristics of the patient's original lesion. However, the existing primary tumor cell culture technologies mainly include 2D culture, 3D culture, reprogramming culture, etc., and these methods are different to varying degrees. Facing problems such as extremely long culture cycle, low culture success rate, difficult removal of miscellaneous cells, and inability to reproduce the tumor microenvironment

Method used

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  • Culture method of gastric cancer micro-tumor cell model
  • Culture method of gastric cancer micro-tumor cell model
  • Culture method of gastric cancer micro-tumor cell model

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0092] Example 1. Preparation of reagents for culturing gastric cancer micro-tumor models

[0093] 1. Sample preservation solution (100mL)

[0094] The specific formulation of the sample preservation solution (100mL) is shown in Table 1.

[0095] Table 1. Sample Preservation Solution (100mL)

[0096]

[0097] 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.

[0098] 2. Sample cleaning solution (100mL)

[0099] The specific formula of the sample cleaning solution (100mL) is shown in Table 2.

[0100] Table 2. Sample cleaning solution (100mL)

[0101]

[0102] The sample cleaning solution should be prepared and used immediately.

[0103] 3. Sample dissociation solution (10mL)

[0104] The specific formulation of the sample dissociation solution (10mL) is shown in Table 3.

[0105] Table 3. Sample dissociation solution (10mL)

[0106]

[0107]

[...

Embodiment 2

[0206] Example 2, acquisition of postoperative specimens of gastric cancer

[0207] 1. Cooperate with tertiary hospitals to obtain samples through the form of clinical research initiated by researchers, and the cooperation has passed the formal medical ethics review.

[0208] 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 for the samples are: primary gastric cancer, and the pathological stage is stage II or In stage III, gastric cancer samples weighing more than 20mg.

[0209] 3. All enrolled cases are uniformly coded by the date of sample collection + the last four digits of the patient’s hospital number. For example, for a sample provided on January 1, 2020, the patient’s hospital number is T001537474, and the sample experiment number is 202001017474. Hide the patient...

Embodiment 3

[0212] Embodiment 3, pre-dissociation treatment of gastric cancer tissue samples

[0213] The following operations need to be performed on ice, and the entire operation steps need to be completed within 10 minutes.

[0214] The surgical equipment used in the following operations must be sterilized by high-temperature steam (120°C, 20 minutes) in advance and dried before use.

[0215] 1. After the sample is weighed, clean the surface of the sample with medical alcohol (75% by volume) for 10 to 30 seconds.

[0216] 2. Wash the sample 5 times with sample cleaning solution, and wash the sample 5 times with sterile PBS solution.

[0217] 3. 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 a gastric cancer micro-tumor cell model. The invention provides a novel gastric cancer micro-tumor model culture technology and a matched reagent, and the core of the technology is as follows: (1) using a mild cell dissociation reagent for treating gastric cancer solid tumor tissues, so that the vitality of various types of cells in the tissues is ensured to the greatest extent; and (2) preparing a special serum-free culture medium, and self-assembling various types of cells separated from gastric cancer tissues by using a suspension culture system to form a cell mass structure with various cell components, which is called as the gastric cancer micro-tumor model. The gastric cancer micro-tumor model obtained by the method can accurately reflect various characteristics of the original focus of a patient, and is a good scientific research experiment model and a preclinical experiment model in the field of precise tumor diagnosis and treatment. It can be foreseen that the culture method has wide application prospects in the fields of gastric 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 gastric cancer micro-tumor cell models. Background technique [0002] Gastric cancer is one of the most common malignant tumors that seriously threaten human health. The current treatment methods for gastric cancer mainly include surgery, chemotherapy, targeted therapy, and immunotherapy. However, the risk of recurrence and metastasis of gastric cancer is high, and more than 50% of patients with gastric cancer will experience different degrees of recurrence and metastasis within months to years after radical treatment. [0003] In the past 10 years, targeted therapy and immunotherapy based on next-generation gene sequencing technology (NGS) have completely changed the treatment pattern of malignant tumors. New drugs and new diagnosis and treatment techniques have improved the prognosis of gastric cancer patients to a certain extent, but targeted drugs and imm...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/09C12N5/071
CPCC12N5/0693C12N5/0679C12N2500/32C12N2501/11C12N2501/115C12N2501/12C12N2501/415C12N2501/155C12N2501/2302C12N2501/2315C12N2501/727C12N2501/15C12N2500/30C12N2500/25C12N2501/345C12N2509/10C12N2509/00
Inventor 尹申意张函槊
Owner SUZHOU GENOARRAY
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