In vitro culture and characterization of a mammary epithelial cell line from Chinese Holstein dairy cow.
BACKGROUND
The objective of this study was to establish a culture system and elucidate the unique characteristics of a bovine mammary epithelial cell line in vitro.


METHODS
Mammary tissue from a three year old lactating dairy cow (ca. 100 d relative to parturition) was used as a source of the epithelial cell line, which was cultured in collagen-coated tissue culture dishes. Fibroblasts and epithelial cells successively grew and extended from the culturing mammary tissue at the third day. Pure epithelial cells were obtained by passages culture.


RESULTS
The strong positive immunostaining to cytokeratin 18 suggested that the resulting cell line exhibited the specific character of epithelial cells. Epithelial cells cultured in the presence of 10% FBS, supraphysiologic concentrations of insulin, and hydrocortisone maintained a normal diploid chromosome modal number of 2n=60. Furthermore, they were capable of synthesizing beta-casein (CSN2), acetyl-CoA carboxylase-alpha (ACACA) and butyrophilin (BTN1A1). An important finding was that frozen preservation in a mixture of 90% FBS and 10% DMSO did not influence the growth characteristics, chromosome number, or protein secretion of the isolated epithelial cell line.


CONCLUSIONS
The obtained mammary epithelial cell line had normal morphology, growth characteristics, cytogenetic and secretory characteristics, thus, it might represent an useful tool for studying the function of Chinese Holstein dairy cows mammary epithelial cell (CMECs).
Hu
Han
H
State Key Laboratory of Animal Nutrition, Institute of Animal Science, Chinese Academy of Agricultural Science, Beijing, China.
Wang
Jiaqi
J
Bu
Dengpan
D
Wei
Hongyang
H
Zhou
Linyun
L
Li
Fadi
F
Loor
Juan J
JJ
D000818:Animals; D002417:Cattle; D018929:Cell Culture Techniques / Q000379:methods; D002460:Cell Line; D049109:Cell Proliferation; D002681:China; D003094:Collagen / Q000737:chemistry; D003470:Culture Media / Q000378:metabolism; D004847:Epithelial Cells / Q000166:cytology*; D005260:Female; D005347:Fibroblasts / Q000166:cytology; D007621:Karyotyping; D053538:Keratin-18 / Q000096:biosynthesis; D008321:Mammary Glands, Animal / Q000166:cytology*
D003470:Culture Media; D053538:Keratin-18; D003094:Collagen