A "selective secondary tissue attachment" method for isolation and purification of mammary epithelial cells.

ABSTRACT

BACKGROUND: Mammary epithelial cells (MECs) are specific target cells to study the underlying mechanism for mammary gland development and lactation, mammary gland bioreactor and breast cancer pathogenesis. However, the conventional isolation and purification methods of MECs have restricted the research and application of its related fields, because the isolated MECs population was always mixed with other cell types. OBJECTIVE: In order to solve the problem of incomplete purification of MECs isolated in vitro, it is very necessary to establish the optimal culture method for its isolation and purification. METHOD: In this study, a method were called the "selective secondary tissue attachment method", which were proved to be effective and accurate by the purity, cellular viability, biological characteristic, and transgenic efficiency of the purified MECs. KEY RESULTS: The results showed that compared with traditional methods, this method could obtain 100% MECs population with high purity and high cell viability in vitro, and these purified cells showed high transgenic efficiency and a high number of positive clones. In addition, the early embryo development rate could significantly improve when the purified mammary epithelial cells were used as donor cells for nuclear transfer. CONCLUSIONS: Therefore, this study provides a versatile and effective method for the isolation of MECs with high purity, which can be used further for mammary-related research. IMPLICATIONS It is most effective approach to isolate mammary epithelial cells by "selective secondary tissue attachment method".