title: β-casein gene expression by in vitro cultured bovine mammary epithelial cells derived from developing mammary glands.
abstract: Epithelial cells from mammary gland tissue that are cultured in vitro are able to maintain specific functions of this gland, such as cellular differentiation and milk protein synthesis. These characteristics make these cells a useful model to study mammary gland physiology, development and differentiation; they can also be used for production of exogenous proteins of pharmaceutical interest. Bovine mammary epithelial cells were cultured in vitro after isolation from mammary gland tissue of animals at different stages of development. The cells were plated on Petri dishes and isolated from fibroblasts using saline/EDTA treatment, followed by trypsinization. Cells isolated on plastic were capable of differentiating into alveolus-like structures; however, only cells derived from non-pregnant and non-lactating animals expressed β-casein. Real-time qPCR and epifluorescence microscopy analyses revealed that alveolus-like structures were competent at expressing Emerald green fluorescent protein (EmGFP) driven by the β-casein promoter, independent of β-casein expression.
authors.affiliation: Faculdade de Zootecnia e Engenharia de Alimentos da Universidade de São Paulo, Pirassununga, SP, Brasil. monzani.paulo@gmail.com
authors.forename: P S
authors.initials: PS
authors.lastname: Monzani
authors.forename: F F
authors.initials: FF
authors.lastname: Bressan
authors.forename: L G
authors.initials: LG
authors.lastname: Mesquita
authors.forename: J R
authors.initials: JR
authors.lastname: Sangalli
authors.forename: F V
authors.initials: FV
authors.lastname: Meirelles
mesh_terms: D000818:Animals; D002364:Caseins / Q000096:biosynthesis* / Q000235:genetics* / Q000378:metabolism; D002417:Cattle; D002454:Cell Differentiation; D002478:Cells, Cultured; D004847:Epithelial Cells / Q000166:cytology* / Q000378:metabolism; D005260:Female; D015870:Gene Expression; D049452:Green Fluorescent Proteins; D007774:Lactation / Q000502:physiology; D016086:Lentivirus / Q000235:genetics; D008321:Mammary Glands, Animal / Q000166:cytology / Q000196:embryology*; D008894:Milk Proteins / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism; D016133:Polymerase Chain Reaction; D011247:Pregnancy; D011401:Promoter Regions, Genetic
chemical_list: D002364:Caseins; D008894:Milk Proteins; D049452:Green Fluorescent Proteins