title: Isolation, characterization, and EGFP expression in the buffalo (Bubalus bubalis) mammary gland epithelial cell line.
abstract: This study was aimed to establish a buffalo mammary epithelial cells (BuMECs) line and maintain it for long-term by subculturing. BuMECs isolated from lactating buffalo mammary glands were cultured on a collagen matrix gel. BuMECs expressed significant amounts of the epithelial cell specific marker cytokeratin 18 as determined by immunohistochemistry. The BuMECs displayed monolayer, cobble-stone morphology, and formed lumen-, dome-, and duct-like structures. Furthermore, they were capable of synthesizing CSN2, BLG, ACACA, and BTN1A1, showed viability after thawing and expressed milk protein genes. The enhanced green fluorescent protein gene was transferred successfully into the BuMECs using lipofection method and the transfected cells could be maintained for long-term in culture by subculturing.
authors.affiliation: Embryo Biotechnology Lab, Animal Biotechnology Centre, National Dairy Research Institute, Karnal, 132001, India.
authors.forename: Ramakant
authors.initials: R
authors.lastname: Kaushik
authors.forename: Karn Pratap
authors.initials: KP
authors.lastname: Singh
authors.forename: Archana
authors.initials: A
authors.lastname: Kumari
authors.forename: Manoj Kumar
authors.initials: MK
authors.lastname: Singh
authors.forename: Radhey Shyam
authors.initials: RS
authors.lastname: Manik
authors.forename: Prabhat
authors.initials: P
authors.lastname: Palta
authors.forename: Suresh Kumar
authors.initials: SK
authors.lastname: Singla
authors.forename: Manmohan Singh
authors.initials: MS
authors.lastname: Chauhan
mesh_terms: D000818:Animals; D002020:Buffaloes / Q000378:metabolism*; D018929:Cell Culture Techniques; D002460:Cell Line; D017931:DNA Primers / Q000235:genetics; D004847:Epithelial Cells / Q000166:cytology / Q000378:metabolism*; D005260:Female; D049452:Green Fluorescent Proteins / Q000378:metabolism*; D007150:Immunohistochemistry; D053538:Keratin-18 / Q000378:metabolism; D007774:Lactation / Q000502:physiology; D008321:Mammary Glands, Animal / Q000166:cytology*; D008894:Milk Proteins / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection
chemical_list: D017931:DNA Primers; D053538:Keratin-18; D008894:Milk Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins