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A method for isolating and culturing human meningioma cells

A technology for meningioma and human brain, applied in the field of biomedicine, can solve the problems of inability to obtain a large number of living cells, slow cell growth, and slow cell growth rate.

Active Publication Date: 2015-09-30
AFFILIATED HUSN HOSPITAL OF FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, the factors that make it difficult to obtain a large number of primary and subcultured meningioma cells are as follows: (1) The interstitial cells of meningioma tissue include many stroma / fibers, and it is difficult to digest meningioma tissue by mechanical separation or single enzyme method , hard mechanical separation or enzyme digestion for a long time, resulting in cell fragmentation, so a large number of living cells cannot be obtained from the tissue; (2) Since most meningiomas are benign, the cells grow slowly and cannot crawl to the cell-free cells far away from the tissue block Zone growth, can only climb out and grow around the tissue block
Therefore, it is difficult for the cells to cover the bottom of the culture bottle; (3) due to the slow growth rate of the cells, when the monolayer cells are adhered to the wall for culture, it is difficult to pass on for a long time with conventional cell seeding density; (4) a suitable method has not been established. The culture system of meningioma cells, the growth factors or hormones that can promote the proliferation of meningioma cells are not clear

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  • A method for isolating and culturing human meningioma cells
  • A method for isolating and culturing human meningioma cells
  • A method for isolating and culturing human meningioma cells

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Embodiment 1

[0042] 1 Materials and methods:

[0043] 1.1 Reagents are all cell culture grade reagents, unless otherwise specified are GIBCO products.

[0044] 1.2 The materials come from 6 isolated meningioma tissues excised from neurosurgery patients with meningioma in Huashan Hospital Affiliated to Fudan University.

[0045] 1.3 Method

[0046] (1) Separation of primary human meningioma cells:

[0047] a. The isolated human meningioma tissue was aseptically collected, and the surgically resected meningioma tissue was washed more than 5 times in ice-cold HBSS.

[0048] b. Cut the tissue with scissors, incubate the tissue in PBS containing Dispase / Collegnase and 0.04% DNase at 37°C for 30 minutes, blow and suck more than ten times with a Pasteur pipette, then vertically rest the centrifuge tube for 5 minutes, and collect the upper layer Digest the suspension and collect the digested cells by centrifugation;

[0049] c. Add an appropriate amount of enzyme to the undigested tissue in th...

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Abstract

The invention belongs to the biomedical field and relates to a method for isolating and culturing meningioma cells and in particular relates to a method for isolating meningioma cells from isolated human meningioma tissues and subculturing the meningioma cells. The method is characterized by adopting a cocktail type combined enzyme to digest the isolated human meningioma tissues stepwise to obtain meningioma cell suspension and carrying out low-oxygen culture on the meningioma cell suspension in a specific culture solution to obtain primary meningioma cells; and carrying out suspension culture on the obtained primary meningioma cells in the specific culture solution under the low-oxygen condition until more than 15 generations of meningioma cells are obtained. The method has the advantages that the cocktail type combined enzyme and the stepwise digestion method are beneficial to preserving cell viability, so that the cells obtained through digestion are basically viable cells, thus providing cell sources for primary culture of meningioma cells; and the method is beneficial to isolation and proliferation of meningioma cells and provides cell sources for further carrying out meningioma mechanism and drug screening researches.

Description

technical field [0001] The invention belongs to the field of biomedicine and relates to a method for cell separation and culture, in particular to a method for the separation and culture of meningioma cells, in particular to a method for separating and culturing meningioma cells from isolated human meningioma tissues. Background technique [0002] According to reports, meningioma is a common intracranial tumor, accounting for about 20% of the total number of intracranial tumors, and the incidence rate is higher in women. Studies have shown that most meningiomas are benign, with a long course of disease, and their distribution is roughly similar to that of arachnoid granules, usually in the parasagittal sinus of the cerebral hemisphere. Surgical resection is the preferred clinical treatment method. Treatment practices show that some meningiomas have a higher recurrence rate 5 years after resection; and some tumors are difficult to achieve total surgical resection because of t...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/09
Inventor 宫晔沙红英汤海亮谢清郑名哲汪戴军陈衔城毛颖周良辅
Owner AFFILIATED HUSN HOSPITAL OF FUDAN UNIV
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