Characteristics and EGFP expression of porcine mammary gland epithelial cells.
The aims of this study were to establish a porcine mammary gland epithelial (PMGE) cell line, and to determine if these PMGE cells could be maintained long-term in culture by continuous subculturing following transfection with a reporter gene, enhanced green fluorescence protein (EGFP). Primary culture of PMGE cells was achieved by outgrowth of migrating cells from the fragments of the mammary gland tissue of a lactating pig. The passage sixteen PMGE cells were transfected with EGFP gene using lipofection. The expression of Cell keratins of epithelial cells in PMGE cells was tested by immunofluorescence. Βeta-Casein gene mRNA was tested for PMGE cells by RT-PCR. The results showed that PMGE cells could form dome-like structure which looked like nipple, and the cells contained different cell types. The expression of Cell keratins demonstrated the property of epithelial cells, and the PMGE cells could express transcript encoding a Βeta-Casein protein. EGFP gene was successfully transferred into the PMGE cells, and the transfected cells could be maintained long-term in culture by continuous subculturing. In conclusion, we have established a EGFP gene transfected porcine mammary gland epithelial (ET-PMGE) cell line.
Zheng
Yue-Mao
YM
College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi 712100, China. zhengyuemao@163.com
He
Xiao-Ying
XY
D000818:Animals; D002364:Caseins / Q000235:genetics; D002478:Cells, Cultured; D004848:Epithelium / Q000378:metabolism; D005260:Female; D005455:Fluorescent Antibody Technique / Q000662:veterinary; D005786:Gene Expression Regulation / Q000502:physiology; D005796:Genes / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000096:biosynthesis*; D008321:Mammary Glands, Animal / Q000166:cytology / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000662:veterinary; D013552:Swine; D014162:Transfection / Q000662:veterinary
D002364:Caseins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins