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Human intestinal cancer primary cell and application and culture method thereof

A technology of primary cells and culture methods, applied in the field of human intestinal cancer primary cells, application and culture, which can solve problems such as death, poor cell state, and culture failure of human intestinal cancer primary cells

Active Publication Date: 2020-03-17
BEIJING HARMONY HEALTH MEDICAL DIAGNOSTICS CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this method may cause poor cell status or death due to over-digestion, resulting in failure of primary human intestinal cancer cell culture

Method used

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  • Human intestinal cancer primary cell and application and culture method thereof
  • Human intestinal cancer primary cell and application and culture method thereof
  • Human intestinal cancer primary cell and application and culture method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0075] Example 1: Isolation and culture of human intestinal cancer primary cells

[0076] (1) With the informed consent of the patient or the patient's guardian, collect part of the intestinal cancer tissue removed during the patient's operation;

[0077] (2) Transfer the intestinal cancer tissue collected in step (1) to a cell culture dish, wash with a cleaning solution (containing 0.4 mg / ml penicillin, 0.4 mg / ml kanamycin sulfate, 0.4 μg / ml amphotericin B, 4 μg / ml vancomycin in normal saline) washed 8 times to remove non-cancerous tissue impurities such as fat and mucous membrane;

[0078] (3) Transfer the cleaned intestinal cancer tissue in step (2) to a new culture dish, cut the cleaned intestinal cancer tissue into small pieces with scissors and a blade, and chop them into fine pieces;

[0079] (4) Transfer the crushed intestinal cancer tissue to a 50ml centrifuge tube, centrifuge at 2000rpm for 8min, remove the supernatant, add 9ml DF medium for resuspension, and then a...

Embodiment 2

[0084] Example 2: Purification of Human Intestinal Cancer Primary Cells

[0085] (1) When the confluence of the cells cultured in step (8) of Example 1 reaches 70-90%, the serum-free medium is removed and digested with 1-2 ml EDTA-Trypsin.

[0086] (2) Observe under a microscope until the cells are detached from the culture flask, discard the fibroblasts digested by EDTA-Trypsin first, and then collect the digested cells while digesting, and stop the digestion of the collected cells with DF10 medium until All cells were digested to completion.

[0087] (3) Centrifuge at 1500rpm for 6min, remove the supernatant, resuspend with serum-free medium, and store at 37°C, 5% CO 2Place in the cell incubator for 1.5 hours, after the fibroblasts adhere to the wall, collect the unattached cells, transfer them to a new culture bottle to continue culturing, and repeat the adhesion for 3 times to achieve the effect of completely removing the fibroblasts. The primary human intestinal cancer ...

Embodiment 3

[0089] Embodiment 3: Morphological observation

[0090] By observing under an inverted microscope, it can be found that the human intestinal cancer primary cells cultured after Examples 1 and 2 are in the form of epithelial cells. Cells proliferate rapidly, without contact inhibition, squeeze each other, and appear stacked or mosaic. For specific results, see figure 1 .

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PUM

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Abstract

The invention provides a human intestinal cancer primary cell and an application and culture method thereof. The human intestinal cancer primary cell is named as HCOC-691. The preservation number is CGMCC NO. 18528. The preservation address is Beijing of China. The human intestinal cancer primary cell disclosed by the invention can be applied to pathogenesis research of cancer-related diseases, drug sensitivity research and detection and antitumor drug research. Compared with the tumor primary cells cultured by the existing purification culture process, the human intestinal cancer primary cells have the better cell state.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to primary human intestinal cancer cells, application and culture method. Background technique [0002] Intestinal cancer is a common digestive tract disease. Generally, the biological characteristics of primary cultured tumor cells have not changed much because the tissue has just been isolated from the body, and the original genetic characteristics are still retained. The gene retention is more than 90%. It is suitable for biochemical molecular Experimental and drug susceptibility tests and related experiments to explore the mechanism, the data are more convincing. The establishment of primary human intestinal cancer cells is of great significance for the establishment of cell lines, drug sensitivity testing experiments, personalized treatment, and research on the pathogenesis and therapeutic mechanism of intestinal cancer. [0003] At present, primary human intestinal cancer cells a...

Claims

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Application Information

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IPC IPC(8): C12N5/09C12Q1/02C12R1/91
CPCC12N5/0693C12N2500/25C12N2500/32C12N2501/11C12N2501/39C12N2501/998G01N33/5011G01N33/5044
Inventor 李春凤赵九娥智慧芳倪君君
Owner BEIJING HARMONY HEALTH MEDICAL DIAGNOSTICS CO LTD
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