Human breast cancer cell line SK-3rd and construction method thereof

A technology of human breast cancer cells and human breast cancer, which is applied in the establishment of human breast cancer cell lines and the field of line establishment, which can solve the problems of the small proportion of cancer stem cells and the lack of platforms for cancer stem cell research.

Inactive Publication Date: 2008-08-13
SUN YAT SEN UNIV
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Problems solved by technology

[0004] However, since the discovery of breast cancer stem cells in 2003, there have been very few literatures on breast cancer stem cells in the world. Establishment of Human Breast Cancer Cell Lines Without Enrichment of Breast Cancer Stem Cells

Method used

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  • Human breast cancer cell line SK-3rd and construction method thereof
  • Human breast cancer cell line SK-3rd and construction method thereof
  • Human breast cancer cell line SK-3rd and construction method thereof

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Embodiment Construction

[0032] The specific method of the human breast cancer cell line enriching breast cancer stem cells established by the present invention is as follows:

[0033] 1. Establish human breast cancer NOD / SCID mouse orthotopic xenograft model according to conventional methods.

[0034]Breast cancer cell line SK-BR-3 cells were cultured on the wall, the culture medium was DMEM containing 10% (v / v) fetal bovine serum, observed under an inverted microscope, when the cells were in the logarithmic growth phase, the culture medium was removed and rinsed with PBS Once, add 2ml of 0.25% trypsin, observe under the microscope, add 4ml of serum-containing culture medium to stop the digestion after the cells become round, blow gently with a pipette, transfer to a centrifuge tube, centrifuge at 1000rpm and resuspend in PBS to prepare single cells Suspension, counted using a counting plate, adjusted the cell concentration, so that the inoculation volume was 100 μl, a total of 2 × 10 6 Cells were i...

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Abstract

Human breast carcinoma SK-BR-3 cell strain as source cell for building train adapted by the invention is continuously inoculated on female NOD / SCID mouse mammary gland fat pad and filtered in pressure of low dose chemotherapy drug tail intravenous injection to build human breast carcinoma cell line SK-3rd rich in tumour breast carcinoma stem cell. The proper of knub stem cell in the cell is increased greatly comparing with non NOD / SCID vivo passage source cell. The formation rate of saccule and the number of cell in the saccule are increased greatly in case of cell suspension culture. The ratio of undifferentiation cancer cell is reduced but is higher evidently than source cell in process of SK-3rd saccule cell inducing differentiation. The saccule cell formed from SK-3rd cell has carcinoma stem cell phenotype which gradually dies out in process of differentiation. NOD / SCID mouse orthotopic transplantation has high tumour-forming rate, high long-distance transferring rate, short tumour-forming periodic time and less number of cells needed by tumour-forming. The invention is an ideal mould for studying breast carcinoma stem cell and has wide applications foreground for disclosing generation mechanism of breast carcinoma and neoplastic treat.

Description

technical field [0001] The invention belongs to the field of microbial animal cell lines, and in particular relates to the establishment of a human breast cancer cell line enriched with breast cancer stem cells and a method for establishing a line. Background technique [0002] In recent years, more and more studies have proved that there are cancer cells with different degrees of differentiation and tumorigenic ability in solid malignant tumors, and only a small number of cells have particularly strong tumorigenic ability and extremely low degree of differentiation. And the characteristics of multi-lineage differentiation potential, so it is called "cancer stem cells (cancer stem cells)", or "tumorigenic cancer cells (tumorigenic cancer cells)". Similar to the proliferation characteristics of stem cells, cancer stem cells with strong proliferation potential proliferate slowly, and most cells are in the G phase of the cell cycle. 0 / G 1 stage, and express a large number of...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/08C12N5/09
Inventor 宋尔卫于风燕胡孝渠
Owner SUN YAT SEN UNIV
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