Two breast cancer stem cell lines capable of being stably cultured and keeping original characteristics, separation method and uses thereof

Abstract

The invention discloses two breast cancer stem cell lines capable of being stably cultured and keeping original characteristics, separation method and uses thereof. A breast cancer tumor specimen just cut off from the tumor is taken, then is subjected to collagenase digestion and is filtered; a flow cytometer sorts CD44<+>, CD24<-> and ALDH1<+> cells, the cells are adjusted to a purity of more than 98%; a stem cell culturing agent for breast cancer cell is prepared; the sorted cells are screened in a monoclonal manner and the obtained monoclone cells are inoculated in an ultralow adhesion culture dish containing breast cancer stem cell culturing agents; penicillin and streptomycin are added in the original culturing medium; culturing is performed in CO<2> cultivation box. In the continuous culturing process, the breast cancer stem cell lines capable of being stably cultured and keeping original characteristics keep ball formation ability, self-updating ability, CD44<+>, CD24<-> / low Lin- ALDH1<+> phenotype, pluripotency differentiation ability, in vitro soft agar tumorigenesis ability, in vivo tumorigenesis ability in nude mouse, and in vivo tumorigenesis pluripotency differentiation ability in nude mouse.

Description

Technical field: [0001] The invention belongs to the field of medicine and biology, and specifically relates to two breast cancer stem cell lines that can be stably passed down and maintain the original characteristics, their separation method and application. Background technique: [0002] Breast cancer is the most common malignant tumor in women, and its incidence rate is increasing year by year. According to statistics from the American Cancer Society (ACS) in 2012, there were 230,000 newly diagnosed cases in the United States, an increase of 29%, and 40,000 deaths, an increase of 14%. With the change of lifestyle in my country, the acceleration of urbanization, and the promotion of mammography, the number of newly detected cases increases year by year. The incidence of breast cancer in Shanghai, China, rose from 17 / 100,000 in 1972 to 2007. 69.22 / 100,000. At the same time, the incidence of breast cancer in China is younger than that in Western countries. The peak age of...

AI Technical Summary

Problems solved by technology

However, as far as we know, no laboratory has established a clinically derived breast cancer stem cell line that can be stably passaged. Therefore, there is an urgent need to establish a clinically derived breast cancer stem cell line that can be stably passaged and used in clinical and scientific research

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