Isolation and Culture of Primary Human Mammary Epithelial Cells

生物 细胞培养 乳腺 体外 原代培养 乳腺癌 细胞生物学 化学定义介质 免疫学 癌症研究 癌症 遗传学
作者
Linfeng Qian,J. Stephen Yan,Shiqi Chen,Maryam Mohanmmed Abbas Karekad,Yue Wu,Xunwei Wu,Xiaohong Xu,Xiaoqun Zhang
出处
期刊:Journal of Visualized Experiments [MyJoVE Corporation]
卷期号: (207)
标识
DOI:10.3791/66638
摘要

The mammary gland is a fundamental structure of the breast and plays an essential role in reproduction. Human mammary epithelial cells (HMECs), which are the origin cells of breast cancer and other breast-related inflammatory diseases, have garnered considerable attention. However, isolating and culturing primary HMECs in vitro for research purposes has been challenging due to their highly differentiated, keratinized nature and their short lifespan. Therefore, developing a simple and efficient method to isolate and culture HMECs is of great scientific value for the study of breast biology and breast-related diseases. In this study, we successfully isolated primary HMECs from small amounts of mammary tissue by digestion with a mixture of enzymes combined with an initial culture in 5% fetal bovine serum-DMEM containing the Rho-associated kinase (ROCK) inhibitor Y-27632, followed by culture expansion in serum-free keratinocyte medium. This approach selectively promotes the growth of epithelial cells, resulting in an optimized cell yield. The simplicity and convenience of this method make it suitable for both laboratory and clinical research, which should provide valuable insights into these important areas of study.
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