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Separation and culture method of primary human breast cancer cells

A technology for human breast cancer cells and breast cancer cells, which is applied in the field of cell biology and can solve problems such as the difficulty of isolating and purifying human breast cancer CAFs

Inactive Publication Date: 2018-05-25
JIANGYIN CHI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, human breast cancer contains a large amount of adipose tissue, which makes it difficult to isolate and purify human breast cancer CAF.

Method used

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  • Separation and culture method of primary human breast cancer cells

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Embodiment Construction

[0013] In order to demonstrate the purpose and advantages of the present invention more clearly, specific embodiments are now further described. The specific embodiments described here are only for explaining the present invention, and are not intended to limit the present invention.

[0014] The invention selects fresh human breast cancer tissue and separates human breast cancer cells. The specific operation is as follows:

[0015] 1. Rinse tissue specimens 2-3 times with pre-cooled PBS containing double antibodies;

[0016] 2. Fully shred the tissue specimen, add 0.05%-0.1% type I collagenase, 0.05%-0.1% type II collagenase and 0.1%-0.15% hyaluronidase mixed enzyme solution, shake at 37°C Digest in a water bath for 1.5h-2h;

[0017] 3. Stop the digestion with the mixed complete medium containing 10% FBS H-DMEM and RPMI1640 (2:1);

[0018] 4. Filter the digested tissue through 80-mesh, 100-mesh, and 200-mesh sieves in turn, collect the filtrate, centrifuge at 1800rpm / min ...

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Abstract

The invention provides a separation and culture method of primary human breast cancer cells. The separation and culture method comprises the following steps: (a) washing a tissue sample by utilizing adouble-antibody-containing pre-cooling PBS (Phosphate Buffer solution); (b) shearing tissues and digesting a mixed enzyme solution; (c) completely mixing a culture medium and stopping digestion; (d)continuously filtering through a screen mesh and collecting filtrate and centrifuging; (e) re-suspending cells and precipitating, and inoculating into a culture bottle; (f) after culturing for a few days, replacing the completely mixed culture medium and continually culturing. The method for obtaining the human breast cancer cells provided by the invention has short time consumption and high efficiency.

Description

technical field [0001] The invention belongs to the field of cell biology, and in particular relates to a method for separating and culturing primary human breast cancer cells. Background technique [0002] Breast cancer is a malignant tumor that usually occurs in the glandular epithelial tissue of the breast. It is one of the most common malignant tumors that seriously affects women's physical and mental health and even threatens their lives. The occurrence and metastasis of tumors are closely related to the microenvironment of tumor cells . The tumor microenvironment is an intricate system, and cancer-associated fibroblast (CAS) is one of the most important host cells in the tumor microenvironment. Primary cultured human breast cancer CAFs are directly isolated from tissues, and their biological and genetic characteristics have not changed significantly. They are closest to and reflect their growth characteristics in vivo. It is the best way to study the biological charac...

Claims

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Application Information

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IPC IPC(8): C12N5/09
CPCC12N5/0693C12N2509/00
Inventor 不公告发明人
Owner JIANGYIN CHI SCI