Separation and culture method for different cellular components of human mammary tissue

A technology of breast tissue and culture method, which is applied in the field of cell biology and biology, can solve the problems of limiting the construction and research of multi-layer cell models, and achieve the effect of simple separation and culture methods and good cell models

Inactive Publication Date: 2015-04-01
GUANGDONG OCEAN UNIVERSITY
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Problems solved by technology

However, most of the cell models used in breast cancer research in the past are only for breast cancer epithelial cells, and there are few studies on the influence of other cell components in other breast tissues on the occurrence and development of breast cancer. One of the reasons is that the current iso

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  • Separation and culture method for different cellular components of human mammary tissue
  • Separation and culture method for different cellular components of human mammary tissue
  • Separation and culture method for different cellular components of human mammary tissue

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Embodiment Construction

[0026] The present invention will be further described in detail by examples below, and these examples are only used to illustrate the present invention, and do not limit the scope of the present invention.

[0027] The isolation and culture process of different cell components of breast tissue is as follows:

[0028] (1) Material: fresh breast cancer tissue obtained from a tumor resection operation in a cancer hospital (female, 48 years old).

[0029] (2) Isolation and culture step: breast cancer tissue was placed in phenol red-free DMEM / F12 (1:1) culture solution, preserved at 4°C and transported to the laboratory on the same day. Use surgical scissors in the sterile room to remove blood clots, blood vessels and fibrous tissues in the breast tissue, soak them in 75% alcohol for 30 seconds, then wash them three times with DMEM / F12 (1:1) culture solution, and cut them. crushed, placed in a Petri dish, and added with a digestion medium equivalent to 5-10 times the volume of ma...

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Abstract

The invention relates to a separation and culture method for different cellular components of a human mammary tissue, and belongs to the technical field of cell culture. The method comprises the following steps: (1) sampling a fresh human mammary tissue specimen; (2) carrying out mechanical shearing and collagenase and hyaluronidase combined digestion to prepare a mammary tissue cell suspension; (3) carrying out low-speed and differential centrifugation to layer and separate the different cellular components; (4) carrying out centrifugation, washing and adherent culture for multiple times to purify the cell components, so as to obtain the different cellular components of the mammary tissue, which comprise epithelial cells, matrix cells and preadipocytes. The separation and culture method disclosed by the invention can be used for quickly, simply and effectively separating and purifying the different cellular components from the same mammary tissue; the cultured mammary gland cells are sufficient in quantity, good in cell viability and purity which is up to 95% above; the separation and culture method can provide very useful materials for molecular biology study associated with mammary glands and breast cancer, and establishes a foundation for construction of a mammary gland microenvironment multi-cell culture model.

Description

technical field [0001] The invention relates to a method for separating and culturing tissue cells, in particular to a method for separating and culturing different cell components of human breast tissue, and belongs to the field of cell biology and biotechnology. Background technique [0002] Breast cancer is a malignant tumor that usually occurs in breast epithelial tissue. It is one of the most common tumors in women, seriously affecting women's physical and mental health and even threatening their lives. According to WHO statistics, the incidence rate accounts for 7-10% of various malignant tumors in the whole body. About 1.2 million women in the world suffer from breast cancer every year, and 500,000 people die from the disease. [0003] The occurrence and development of tumor is a complex process involving many factors. Under normal conditions, the homeostasis within epithelial tissues is maintained by dynamic interactions between epithelial cells and their microenvir...

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Application Information

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IPC IPC(8): C12N5/071C12N5/077
Inventor 钟赛意叶伟平徐萍萍谌素华秦小明王维民
Owner GUANGDONG OCEAN UNIVERSITY
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