Establishment of an in vitro culture model to study milk production and the blood-milk barrier with bovine mammary epithelial cells

Yusaku Tsugami, Norihiro Suzuki, Manabu Kawahara, Takahiro Suzuki, Takanori Nishimura, Ken Kobayashi

Research output: Contribution to journalArticlepeer-review

Abstract

This study attempted to establish a culture model to recreate the milk production pathway in bovine mammary epithelial cells (BMECs). BMECs were isolated from Holstein cows (nonlactating, nonpregnant, and parous) and were stored by cryopreservation. To separate the apical and basolateral compartments, BMECs were cultured on a cell culture insert with a collagen gel in the presence of bovine pituitary extract and dexamethasone to induce milk production and tight junction (TJ) formation. The culture model showed the secretion of the major milk components, such as β-casein, lactose, and triglyceride, and formed less-permeable TJs in BMECs. Moreover, the TJs were distinctly separated from the apical and basolateral membranes. Glucose transporter-1, which transports glucose into the cytoplasm through the basolateral membrane, localized in the lateral membrane of BMECs. Toll-like receptor-4, which binds to lipopolysaccharide in the alveolar lumen in mastitis, localized in the apical membrane. Beta-casein was mainly localized near the Golgi apparatus and the apical membrane. Moreover, milk components were almost secreted into the upper chamber of the cell culture insert. These findings indicate that this model has clear cell polarity as well as in vivo and is effective to study of milk production and the blood-milk barrier in lactating BMECs.

Original languageEnglish
Pages (from-to)e13355
JournalAnimal science journal = Nihon chikusan Gakkaiho
Volume91
Issue number1
DOIs
Publication statusPublished - Jan 1 2020
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • Agricultural and Biological Sciences(all)

Fingerprint Dive into the research topics of 'Establishment of an in vitro culture model to study milk production and the blood-milk barrier with bovine mammary epithelial cells'. Together they form a unique fingerprint.

Cite this