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Human pituitary adenoma cell line and application thereof

A technology for pituitary adenomas and uses, applied in the field of tumor biology, can solve the problems of pituitary tumor cells without humanization, difficult to transfer, etc.

Inactive Publication Date: 2017-06-13
BEIJING NEUROSURGICAL INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the large species differences between rodent pituitary tumor cells and human pituitary tumor tissue, the experimental data obtained from the mouse pituitary tumor cell platform is still difficult to transfer to human experiments, so mature human pituitary adenoma Cells need to be developed
[0005] At present, there are only mouse pituitary tumor cells, and there are no humanized pituitary tumor cells. There are many difficulties in the primary culture of human pituitary tumor cells

Method used

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  • Human pituitary adenoma cell line and application thereof
  • Human pituitary adenoma cell line and application thereof
  • Human pituitary adenoma cell line and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Embodiment 1: Acquisition and cultivation of human pituitary adenoma cell line

[0041] 1. HPA1446 primary cell culture

[0042] 1. Source of biological material

[0043] The cell line of the present invention is obtained by separating and culturing the pituitary tumor tissue of a non-functioning pituitary tumor 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 a non-functioning pituitary adenoma. Under the microscope, the tumor cells were densely distributed, the boundaries of the cell membrane were unclear, and the size and shape of the nuclei were not consistent. , nucleoli and binuclear tumor cells can be seen.

[0044] 2. Isolation and culture method

[0045] Wash the tissue block twice with PBS solution (1:1) containing red solution (purchased from Gibco) in a sterile ultra-clean bench, remove blood vessels a...

Embodiment 2

[0064] Embodiment 2: Determination of cell growth curve

[0065] 1. Method:

[0066] The primary pituitary adenoma cells of Example 1 and the immortalized pituitary adenoma cells were adjusted to a cell concentration of 5×10 cells after the cells were overgrown. 4 cells / mL, inoculate primary pituitary tumor cells and immortalized pituitary tumor cells in 96-well culture plate, 100 μl per well, 37°C, 5% CO 2 Cultivate in an incubator; add 10 μl of MTT to each well after culturing for 12h, 24h, 36h, 48h, and 72h respectively, at 37°C, 5% CO 2 Cultivate in the incubator and incubate for 3-4 hours in the dark; absorb the liquid in the well, add 200 μl of DMSO, and shake on the shaker at room temperature for 10 minutes; measure the OD value at the same time point with a wavelength of 492nm on a microplate reader, and use the measured OD value analyze.

[0067] 2. Results:

[0068] Such as figure 1 As shown, the immortalized pituitary tumor cells entered the growth stationary p...

Embodiment 3

[0069] Embodiment 3: cell morphology detection

[0070] 1. Method:

[0071] 1. Observe the cell line CGMCC No.12672 with a phase contrast microscope.

[0072] 2. Observing the ultrastructure of the cell line CGMCC No.12672 with a transmission electron microscope:

[0073] When the cells grow to 90% confluence, discard the culture medium, add 3ml 0.1MPBS (pH value 7.4), scrape the cells with a rubber cell scraper, collect the cells, centrifuge (1000rpm / min, 10 minutes), discard the supernatant, Aldehyde / osmium tetroxide pre-fixed for 2 hours, washed 3 times with 0.1M PBS (pH 7.4), 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).

[0074] 2. Results:

[0075] 1. The growth characteristics of the cell line CGMCC No.12672 under the phase-contrast microscope: most of them are fusiform, some are triang...

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Abstract

The invention discloses a human pituitary adenoma cell line and application thereof, and belongs to the field of tumor biology. The collection number of human pituitary adenoma cell line (HPA1446) is CGMCC NO.12672. The human pituitary adenoma cell line has the advantages that the cell line can be cultured in vitro for a long time, the cell characteristic is not changed, and the cell line is a powerful tool for related research of pituitary adenoma; the cell line is a powerful tool for studying tumor disease and related molecule action mechanisms, preparing tumor related animal experiment models, studying, screening and evaluating tumor treatment medicines / methods / diagnosis reagents, developing tumor medicine targets and tumor biological treatment new techniques, and detecting study of related biological engineering.

Description

technical field [0001] The invention relates to a strain of human pituitary adenoma cell and its application, belonging to the field of tumor biology. Background technique [0002] Pituitary adenoma accounts for 10%-15% of primary intracranial tumors and is one of the most common nervous system tumors. It has been reported that the autopsy detection rate of pituitary adenoma is as high as 14.4%, and the MRI detection rate of random population is 22.5%. According to whether it can secrete hormones, pituitary tumors can be divided into hormone-secreting pituitary tumors and non-functioning adenomas. According to the type of hormone secretion, hormone-secreting pituitary adenomas can be further divided into prolactin-, growth-hormone, corticotropin, thyroid-stimulating hormone, and gonadotropin-secreting pituitary adenomas. The clinical manifestations of pituitary tumors are mainly a series of clinical changes caused by tumor space-occupying symptoms and hormone secretion dis...

Claims

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Application Information

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IPC IPC(8): C12N5/09C12Q1/02
CPCC12N5/0693C12N2503/02G01N33/5011G01N2500/10
Inventor 刘潜张亚卓孙异临李储忠
Owner BEIJING NEUROSURGICAL INST
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