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Sorting method for glioma stem cells

A sorting technology for glioma stem cells, which is applied in the field of cell research, can solve the problems of apoptosis and low enrichment efficiency, and achieve the effects of high activity, high purity, and time-saving interference factors

Inactive Publication Date: 2019-08-16
THE FIRST AFFILIATED HOSPITAL OF ARMY MEDICAL UNIV
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Problems solved by technology

The isolation and enrichment efficiency of the suspension culture method is low, and the glioma stem cells in the cell spheres undergo differentiation or spontaneous apoptosis during the continuous passage process

Method used

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  • Sorting method for glioma stem cells
  • Sorting method for glioma stem cells
  • Sorting method for glioma stem cells

Examples

Experimental program
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Embodiment 1

[0029] Embodiment 1, NADH high Sorting of glioma cell subsets and identification of tumor stem cell characteristics

[0030] The main materials and sources used in this example: LN229 was purchased from ACTT (ATCC-CRL-2611), and the primary cultured cells (GBM1) were obtained from clinical glioma samples through primary culture.

[0031] 1. The method for NADH fluorescence determination of glioma cells in fresh glioma samples is as follows:

[0032] (1) According to the WHO classification, 4 samples of grade II glioma, 3 samples of grade III glioma, and 6 samples of grade IV glioma were collected through clinical surgery. Tumor samples (GBM1) were placed on ice, digested with digestive enzymes within 30 minutes and suspended in PBS to make 13 single-cell suspensions, and the cell concentration was adjusted to 1×10 6 cells / mL.

[0033] (2) According to the BD FACSAria II standard procedure of the flow cytometry instrument, start the instrument, stabilize the liquid flow, and...

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Abstract

The invention belongs to the technical field of tumor cell research, and discloses a sorting method for glioma stem cells, specifically, a method for sorting the glioma stem cells by a flow cell sorter according to the autofluorescence property of reduced coenzyme I. The sorting method is free of labelling, simple to operate, low in cost, small in cytotoxicity and good in sorting effect; the dryness of the sorted stem cells is tested, and results show that the glioma stem cells sorted by the method have higher activity and purity than that sorted by current common methods. The establishment ofthe method strongly promotes research on targeted glioma stem cell therapy.

Description

technical field [0001] The invention belongs to the technical field of cell research, and relates to a method for sorting tumor cells, in particular to a method for sorting glioma stem cells. Background technique [0002] Glioma is the malignant tumor of the central nervous system with the highest morbidity and mortality in my country, and glioma stem cells are considered to be the root of its development. Research on the characteristics of glioma stem cells is the basis for establishing targeted glioma stem cell therapy, and the premise of this research is to obtain high activity and high purity glioma stem cells. So far, a variety of technical methods to obtain glioma stem cell-like cells have been established, mainly including surface labeling method, side population (side population, SP) cell method, intracellular enzyme activity (ALDH) method and suspension culture method, but each Both methods have their own limitations. Surface labeling is the most widely used metho...

Claims

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Application Information

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IPC IPC(8): C12N5/095
CPCC12N5/0695C12N2509/00
Inventor 马清华袁野崔有宏崔翔卞修武缪静雅蔡瑞丽王艳霞党微旗
Owner THE FIRST AFFILIATED HOSPITAL OF ARMY MEDICAL UNIV
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