Establishment of Mouse Mammary Epithelial Cell Culture System and Study on the Modulation of Heregulin-α Factor

Abstract

This study amis to establish and perfect the mouse mammary epithelial cell culture system, and to discuss the effect of Heregulin-α (HRG-α) on the mammary epithelial cell proliferation and metabolism within this cell system. It provides important theoretical basis for further research on mammary epithelial cell morphology, structure and function, and reveals the effect and laws of HRG-α in mouse mammary gland development. By comparing the mouse mammary cell growth in different culture media of different pH, serum concentration ratio, we have found that the culture medium of pH 7.4, supplemented with 10% serum was the best conditions for the growth of mammary epithelial cells. Through the use of phase contrast microscopy technology, MTT assay, and other techniques and research methods, this study compared mammary epithelial cell morphology, inoculation survival rate, doubling time, growth curves of biological characteristics at different periods derived. We have found that the good breast cells from the period was the 15 d pregnant mouse mammary. On this basis, important cell marker protein keratin-18 was detected by immunohistochemistry and RT-PCR, to further identify the cells obtained by mammary epithelial cells. Mouse mammary epithelial cells (MMECs) were successfully established <italic>in vitro</italic> culture system. By liquid chromatography and MTT, the study discusses the effects of HRG-α on MMECs in the established cell culture system <italic>in vitro</italic>. The result showes that MMECs’ cell growth and total secretion proteins and lactose were significantly promoted by the appropriate amount of HRG-α. 0.1~20 ng/mL HRG-α could promote MMECs to proliferate and total proteins and lactose content to increase. The maximum value was HRG-α at the concentration of 20 ng/mL. 50 ng/mL, 100 ng/mL HRG-α inhibited the proliferation of MMECs, and decreased the total proteins and lactose content of MMECs.