奶牛乳腺上皮细胞的原代培养及其生物学特性分析

doi:10.11843/j.issn.0366-6964.2022.02.012

Abstract

Abstract: This study aimed to isolate primary bovine mammary epithelial cells (BMECs) from mammary gland tissue and explore its biological characteristics after culture. In this study, the mammary glands of healthy lactating cows were collected from the slaughterhouse, the primary bovine mammary epithelial cells were isolated from the mammary glands by modified enzymatic digestion and verified by morphological observation, immunofluorescence and chromosome karyotype analysis. Meanwhile, the growth curve, population doubling time, resuscitation rate, secretion of milk protein, milk fat, lactose, and mRNA expression of lactation-related genes were investigated at the 3rd, 6th, and 9th generations of mammary epithelial cells. The results showed that the isolated bovine mammary epithelial cells had good purity, and the cell growth presented an S-shaped curve, the population doubling time was 34.87, 41.45 and 65.04 h of the 3 generations of cells respectively, and the resuscitation rate was 88%-93%. In terms of cell secretion function, the secretion of casein, triglycerides, and lactose were detected after induction culture for 2 days, and there were no significant differences among all groups. In addition, all the 3 generations of cells could express the lactation-related genes after induction. This study successfully obtained primary bovine mammary epithelial cells, and proved that the cells still had normal biological functions up to the 9th generation, which could provide experimental materials and technical support for exploring the mechanism of mammary cell proliferation and differentiation in vitro.

Key words: enzymatic digestion, primary culture, biological characteristics, bovine mammary epithelial cells (BMECs)

CLC Number:

S823.91

SUN Peihao, ZHAO Xinzhe, WU Hanxiao, LÜ Ce, YANG Liguo, LIANG Aixin. Primary Culture and Biological Characteristics Analysis of Bovine Mammary Epithelial Cells[J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(2): 447-458.

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Primary Culture and Biological Characteristics Analysis of Bovine Mammary Epithelial Cells

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