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. METHODOLOGY:Mammary tissue from a three year old lactating dairy cow (ca. 100 d relative to parturition) was used as a source of the ep...

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Main Authors: Han Hu, Jiaqi Wang, Dengpan Bu, Hongyang Wei, Linyun Zhou, Fadi Li, Juan J Loor
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2009-11-01
Series:PLoS ONE
Online Access:http://europepmc.org/articles/PMC2768788?pdf=render
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author Han Hu
Jiaqi Wang
Dengpan Bu
Hongyang Wei
Linyun Zhou
Fadi Li
Juan J Loor
author_facet Han Hu
Jiaqi Wang
Dengpan Bu
Hongyang Wei
Linyun Zhou
Fadi Li
Juan J Loor
author_sort Han Hu
collection DOAJ
description 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. METHODOLOGY: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. PRINCIPAL FINDINGS: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).
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spelling doaj.art-a639c6e652e24fc6b45a4e42241b66432022-12-22T01:24:03ZengPublic Library of Science (PLoS)PLoS ONE1932-62032009-11-01411e763610.1371/journal.pone.0007636In vitro culture and characterization of a mammary epithelial cell line from Chinese Holstein dairy cow.Han HuJiaqi WangDengpan BuHongyang WeiLinyun ZhouFadi LiJuan J LoorBACKGROUND: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. METHODOLOGY: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. PRINCIPAL FINDINGS: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).http://europepmc.org/articles/PMC2768788?pdf=render
spellingShingle Han Hu
Jiaqi Wang
Dengpan Bu
Hongyang Wei
Linyun Zhou
Fadi Li
Juan J Loor
In vitro culture and characterization of a mammary epithelial cell line from Chinese Holstein dairy cow.
PLoS ONE
title In vitro culture and characterization of a mammary epithelial cell line from Chinese Holstein dairy cow.
title_full In vitro culture and characterization of a mammary epithelial cell line from Chinese Holstein dairy cow.
title_fullStr In vitro culture and characterization of a mammary epithelial cell line from Chinese Holstein dairy cow.
title_full_unstemmed In vitro culture and characterization of a mammary epithelial cell line from Chinese Holstein dairy cow.
title_short In vitro culture and characterization of a mammary epithelial cell line from Chinese Holstein dairy cow.
title_sort in vitro culture and characterization of a mammary epithelial cell line from chinese holstein dairy cow
url http://europepmc.org/articles/PMC2768788?pdf=render
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