Chordoma cell line and application thereof

A technology for chordoma and its application, which is applied in the field of establishment of human chordoma cell lines, and can solve problems such as unreported

Inactive Publication Date: 2014-01-29
BEIJING NEUROSURGICAL INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The development of targeted drugs first requires a stable chordoma cell line. Based on this chordoma cell line, it is possible to develop drugs that specifically target chordoma. However, such a cell line has not been reported in China so far.

Method used

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  • Chordoma cell line and application thereof
  • Chordoma cell line and application thereof
  • Chordoma cell line and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Embodiment 1: the establishment of human chordoma cell line

[0037] Primary culture of HBC2: the cell line in the present invention is obtained by separating and culturing the skull base chordoma tissue of the surgical patient. The tumor tissue was obtained from a surgical specimen of a patient in the Neurosurgery Center of Beijing Tiantan Hospital Affiliated to Capital Medical University. The pathological diagnosis was brain chordoma. Under the microscope, the cells were arranged in sheets, with round or oval nuclei. Soap-like structure ( figure 1 ). The specific method is as follows: wash the tissue block twice with Hank’s (1:1) containing red solution in a sterile ultra-clean bench, remove blood vessels and necrotic tissue, and cut the tissue block into about 1 mm with tissue scissors 3 Size, add 3ml 0.04% EDTA / 0.5% trypsin, digest in a 37°C water bath for 10 minutes, observe under a microscope, after digesting into single cells, add 300ul fetal bovine serum to st...

Embodiment 2

[0041] Example 2: Observation of Ultrastructure of Cells by Transmission Electron Microscopy

[0042] When the HBC2 cells were subcultured for 14 generations, when the cells reached 90% confluence, discard the culture medium, add 3ml 0.1M PBS (PH7.4), scrape the cells with a rubber cell scraper, collect the cells, centrifuge (800rpm / min, 10 Minutes), discard the supernatant, pre-fix with glutaraldehyde / osmium tetroxide for 2 hours, wash 3 times with 0.1M PBS (pH7.4), 10 minutes each time. Then the specimens were post-fixed, dehydrated, soaked, embedded, cut into ultra-thin sections and stained with heavy metals (completed by the electron microscope room of Beijing Institute of Neurosurgery).

[0043] The ultrastructure of HBC2 was observed by transmission electron microscopy. It can be seen that there are a large number of microvilli on the surface of the cell membrane, the shape of the nucleus is irregular, the nuclear membrane is sunken, the nucleolus is large and obvious, a...

Embodiment 3

[0044] Example 3: Detection of marker proteins derived from notochord tissue by immunohistochemical staining

[0045] When the HBC2 cells were subcultured for 14 generations, after the cells reached 90% confluence, they were washed twice with Hank's. Add 3 ml of 0.04% EDTA / 0.5% trypsin to digest for about 2 minutes, add 2 ml of serum-containing medium to terminate the reaction, centrifuge (800 rpm / min, 10 minutes) and wash once, and collect the precipitated cells at the bottom of the centrifuge tube. Resuspend the cells in 5ml of DMEM / F12 culture medium containing 20% ​​serum, inoculate them on a culture dish covered with coverslips, and discard the culture medium after the cells grow to 90% confluence, and use 0.1M PBS (PH7.4) Wash 3 times, add cold acetone to fix at 4°C for 10 minutes, wash 3 times with 0.1M PBS (PH7.4), 5 minutes each time, 3% hydrogen peroxide for 10 minutes at room temperature (to eliminate endogenous peroxidase). Wash 3 times with 0.1MPBS (PH7.4), 5 min...

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Abstract

The invention discloses a chordoma cell line and application thereof and belongs to the field of oncobiology. The preservation number of a human chordoma cell line (BHC2) is CGMCC (China General Microbiological Culture Collection Center) NO: 5321. The cell line is obtained by separating culture in a chordoma tissue cut from a basicranial chordoma patient operation and is subjected to in vitro subculture for over 14 passages. The cell has the biological characteristics of a notochord-derived cell that the cells are physaliphores arrayed in a cord shape under an optical microscope and a large amount of cavity structures exist in cytoplasm under the transmission of an electron microscope; and the cells expresses various marker proteins of the notochord-derived cell, namely Brachyury, S100, Galectin-3 and Fascin-1. The cell has the tumor characteristic that chromosome is of a heteroploid karyotype; the percentages at each phase of a cell cycle are as follows: 51.3 percent at G1 phase, 23.6 percent at S phase, 25 percent at G2-M phase and the ratio of the G2 to the G1 is approximate to 2, and thus the cell proliferation feature of tumor cells is shown; and the cells obtain tumorigenesis ability after being injected into a nude mouse body. The chordoma cell line can be cultured in vitro for a long term and keeps the cell stability unchanged and is a powerful tool for researching a targeted chordoma tumor medicament.

Description

technical field [0001] The invention relates to a chordoma cell line and its application, belonging to the field of tumor biology, and more specifically relates to the establishment of a human brain chordoma cell line (BHC2) and its application. Background technique [0002] Chordoma is derived from the primary bone tumor produced by the embryonic remnant tissue of the notochord. It is a rare malignant tumor with chronic progression, accounting for 3% to 4% of all primary malignant tumors, ranking fourth among primary malignant bone tumors. bit. About 1 / 3 originate from the base of the skull, and the rest can originate from the spine, sacrum, or other locations than the midline. The tumor develops slowly and has the growth pattern of a malignant tumor. It often grows in the slope bone, the site is deep, and it often grows extensively. Some of them have septations and are adjacent to important blood vessels and nerves. It is very difficult to completely remove them. The tum...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/09C12Q1/02A01K67/027
Inventor 万虹张俊廷冯洁历俊华吴震王亮孙异临郝淑煜李德志
Owner BEIJING NEUROSURGICAL INST
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