Cell line sourced from later-stage gastric signet ring cell cancer, and establishing method and application thereof

A cell line, cancer cell technology, applied in the direction of tumor/cancer cell, microorganism-based method, animal cell, etc. The effect of stable traits, good uniformity and strong clonogenic ability

Inactive Publication Date: 2017-02-01
南京医科大学附属南京医院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] In view of the above-mentioned deficiencies such as the low biodiversity of human gastric cancer cell lines / strains in my country and the large difference in biological characteristics from clinical gastric cancer, the purpose of the present invention is to provide a new cancer cell line gc-006- derived from late gastric signet ring cells. 03

Method used

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  • Cell line sourced from later-stage gastric signet ring cell cancer, and establishing method and application thereof
  • Cell line sourced from later-stage gastric signet ring cell cancer, and establishing method and application thereof
  • Cell line sourced from later-stage gastric signet ring cell cancer, and establishing method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Example 1 Establishment of gc-006-03 of gastric signet ring cell cancer cell line

[0045] The ascites of patients with gastric signet ring cell carcinoma was extracted, and the cells were separated by 1.077g percoll solution. After 1640 culture medium (containing 10% fetal bovine serum, 10mM HEPES, 100U / ml penicillin G, 100μg / ml streptomycin sulfate and 0.25μg / ml amphotericin B), place at 37°C, 5% CO 2 The gastric signet ring cell carcinoma cell line gc-006 was established by in vitro cell subculture in an incubator.

[0046] To establish cell lines: we firstly serially dilute the cells of the gc-006 cell line, inoculate 100-2000 cells / dish in five 10cm culture dishes, use 1640 medium (same as above), place at 37°C, 5% CO 2 Cultured in the incubator for 7-10 days. At this time, the cells are scattered and energetic monoclonal. Then select the monoclonal cells accurately, no need to add trypsin to pick the clones, and directly select the monoclonal cells with the ...

Embodiment 2

[0047] Reality Example 2 Detection of biological characteristics of gastric signet ring cell carcinoma cell line gc-006-03

[0048] 1. Cell morphology:

[0049] (1) Observation of live cells: After the cells have been subcultured and grown stably, they are observed under a phase-contrast microscope. The cells were adhered to the wall and overlapped, without contact inhibition ( figure 1 ).

[0050] (2) HE staining: the cells were passed to 31 passages, and the number of cells reached 10 6 -10 7 Then, paraffin-embedded sections were carried out and observed under the HE staining microscope. The cells showed a malignant epithelioid shape, with a large ratio of nucleoplasm to cytoplasm, multiple nucleoli, and cells with nuclei of different sizes. Mitosis was easily seen, and a certain number of signet ring cells could still be seen ( figure 2 ).

[0051] (3) Observation by transmission electron microscope: few ribosomes in cells, rich in rough endoplasmic reticulum, cyst...

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Abstract

The invention relates to a cell line gc-006-03 sourced from later-stage gastric signet ring cell cancer, and an establishing method and an application thereof. The cell line is preserved in China Center for Type Culture Collection, and is assigned the access number of CCTCC No. C2016152. The cell line gc-006-03 from the gastric signet ring cell cancer allows stable passage. The invention provides certain research clues and experimental models for generation, development and migration of the gastric signet ring cell cancer in future and further disclosure of pathogenesis of the gastric signet ring cell cancer. Meanwhile, establishment of the cell line gc-006-03 not only provides beneficial conditions for molecular biology, immunology and clinical comprehensive diagnosis of the gastric cancer but also provides a convenient platform for screening medicines of preventing and treating the gastric cancer.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a cell line derived from human advanced gastric signet ring cell carcinoma and its establishment method and application. Background technique [0002] Gastric cancer is a malignant tumor of the digestive tract that seriously endangers the health of the people. According to literature reports, there are 989,600 new cases of gastric cancer worldwide, accounting for 8% of the world's new cancer cases; 738,000 deaths related to gastric cancer, accounting for 10% of the world's cancer deaths. More than 70% of new cases and deaths occurred in developing countries. The incidence of gastric cancer in East Asia ranks first in the world. my country is a high-incidence region of gastric cancer, with 300,000 to 400,000 new gastric cancer patients and 300,000 deaths each year, accounting for 23% of the deaths from malignant tumors. In recent years, the age of onset of gastric cancer tends to b...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/09C12Q1/02C12R1/91
Inventor 陈锦飞薛毅琪洪灵芝霍新颖陈红韦栋平
Owner 南京医科大学附属南京医院
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