Isolation and Culture of Human Breast Cancer Cells from Primary Tumors and Metastases

Abstract

Over the years, a number of groups have developed culture media and conditions that support the growth of normal human mammary epithelial cells from a variety of lineages, and these cell systems have increased our understanding of human mammary gland biology. In contrast, it remains difficult to isolate and culture human breast cancer cells from primary tumors and metastatic specimens. Our laboratory has now developed 13 human breast cancer cell lines. Some of these cell lines originated from primary tumor specimens, others were derived from chest wall recurrences in patients who had prior surgery, and some of the cell lines were derived from pleural effusion metastases. The cell lines that we have obtained exhibit an array of genetic alterations representative of the genomic diversity of uncultured human breast cancers. In addition, these cells are cultured under well-defined conditions (most cell lines are grown in serum-free media) that allow detained cellular studies to be carried out that complement the genetic analyses of these cell lines. This chapter outlines the culture methods and approaches that we have used to develop this panel of cell lines. Our methods emphasize certain key features that we feel are essential for the isolation and culture of breast cancer cells. These features are all focused on developing culture conditions that allow for the slow emergence of cancer cells in the relative absence of normal cells, which proliferate rapidly under highly-growth-factor-enriched conditions. Thus, selective growth media and methods for enriching cell populations for cancer cells while minimizing the presence of normal epithelial and stromal elements are important for the expansion of breast cancer cells in vitro. It is still not possible to develop cell lines from every breast cancer specimen obtained in the laboratory. Thus, whereas the cell lines we have developed have resulted in improvements in our understanding of breast cancer cell biology, much work remains to be done to understand why some breast cancer cells fail to grow in vitro while growing robustly in the patient. Improving our knowledge of the factors that influence breast cancer cell viability and proliferation will be important for development of novel therapeutic strategies for breast cancer.