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Vitro model for dynamic modeling human glioma cell displace

A technology of glioma cell and metastasis model, applied in the field of in vitro model of dynamic simulation of human glioma cell metastasis, achieving good reproducibility, high metastatic potential and clear background

Inactive Publication Date: 2008-05-21
SHANDONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

After retrieval, about the construction of an in vitro model for dynamically simulating human glioma cell metastasis and the effective use of the in vitro model for dynamically simulating human glioma cell metastasis to dynamically simulate and analyze the biological activity of transferred glioma cells Systematic research papers and patents have not yet been reported

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  • Vitro model for dynamic modeling human glioma cell displace
  • Vitro model for dynamic modeling human glioma cell displace

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Example 1 Construction of an in vitro model that dynamically simulates the process of human glioma cell metastasis

[0031] 1. Model establishment of anti-cell attachment

[0032] Adherent growth is a prerequisite for the survival of solid tissue cells. Poly-HEMA (purchased from Sigma Company), as an anionic polymer, can be spread on the bottom of the container to prevent the cells from adhering to the wall, so as to construct a cell model in which cells grow in suspension after shedding.

[0033] Dissolve poly-HEMA with 95% ethanol to prepare a solution with a final poly-HEMA concentration of 120mg / ml (i.e. dissolve 2.4g poly-HEMA in 20ml 95% ethanol), shake and mix at 65°C for 8 hours to obtain poly-HEMA HEMA stock solution; then dilute the poly-HEMA stock solution with 95% ethanol at a volume ratio of 1:10 to make a poly-HEMA working solution with a final concentration of 12 mg / ml, store it at 4°C, and set aside; take poly-HEMA Spread the HEMA working solution even...

Embodiment 2

[0045] Changes in the Growth Curve of Glioma Cells in the State of Metastasis in Example 2

[0046] A poly-HEMA-coated 96-well cell culture plate was prepared, and the 96-well cell culture plate was divided into a suspension culture group and an adherent culture group; glioma cells U251 were inoculated into the suspension culture group and the adherent culture group of the 96-well cell culture plate, respectively. In the adherent culture group, 100ul / well, after mixing, bring to 37°C, CO 2 Continue culturing in the incubator for 36 hours, and add 10ul CCK8 to 3 replicate wells of each group every 12 hours, mix well and bring to 37°C, CO 2 The incubation was continued for 90 min in the incubator; the data was read and recorded at 450 nm with a microplate reader, and the growth curve of the glioma cells was drawn according to the data.

[0047] The results showed that the adherent group showed a vigorous proliferation curve, while the suspension group neither proliferated nor a...

Embodiment 3

[0048] Example 3 Cell cycle changes of glioma cells in a metastatic state

[0049] A poly-HEMA-coated 6-well plate was prepared, a metastatic glioma cell group was prepared in the 6-well plate, and a glioma cell group of adherent growth was prepared at the same time. After each group of cells was processed into a single-cell suspension, the cells (including floating cells in the supernatant) were collected, washed, fixed overnight with 70% ethanol, stained with propidium bromide (PI, Sigma product), and detected by flow cytometry. Cell distribution of the cell cycle.

[0050] The results showed that compared with the adherent growth control cells, more cells in the suspension culture group were arrested in G0 / G1 phase (P<0.05).

[0051] The cells in the adherent group and the suspension group were detected by flow cytometry, and the cells with the largest difference were selected for the measurement of the cell cycle at 24h. Proliferative G2 / M phase.

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Abstract

The invention discloses a model in vitro which can simulate the metastasis of human neuroglioma dynamically. The construction steps comprise the construction of an anti-cell adherence model by using human neuroglioma cell line U251; the construction of in vitro metastasis model of human neuroglioma cell and the identification for the constructed metastasis model in vitro of human neuroglioma. The model of the invention can be used for the control research between the in situ cancer and the tumor in metastasis; the invention can further be used as a monitoring system for the biological behavior detection of the tumor cell in metastasis and the judgment of tumor prognosis. The model has important value for the research of neuroglioma metastasis mechanism and the screening for metastasis control drugs. The model can also be used as the screening technology platform for the preparation of anti-neuroglioma transgenic vaccine and the construction of anti-tumor metastasis drugs.

Description

technical field [0001] The invention relates to a human glioma cell metastasis model, in particular to an in vitro model for dynamically simulating human glioma cell metastasis. Background technique [0002] Glioma, also known as glioma, is a variety of tumors that occur in the neuroectoderm, and is the most common in various intracranial tumors. Due to the anatomical characteristics of the nervous system and the head, gliomas are rare Metastasis to other tissues through the vasculature, mostly intracranial. However, most gliomas show expansive-invasive growth, that is, the tumor contains an expansive growth core surrounded by a diffusely invasive marginal zone. Tumors of this growth pattern are often unencapsulated and have no boundary with the surrounding brain tissue. Sometimes it seems to have a clear boundary with the surrounding tissue on the naked eye, but under the microscope, it is found that it often invades the normal brain tissue. The metastasis range of glioma...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/08C12N5/09
Inventor 韩丽辉孙汶生张之勇曹莉莉梁晓红曲忠花杜娟马恬侯楠郭春刘玉刚
Owner SHANDONG UNIV
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