Establishment and characterization of an adherent pure epithelial cell line derived from the bovine oviduct.
The oviduct in vivo has to perform various tasks: maturation and transport of the gametes, milieu preparation for fertilization and embryonic development, and transport of the embryo. The complex arrangement of endocrine and paracrine signals being exchanged between the early embryo and the inner cell layers of the oviduct is barely understood. Therefore, a reproducible, well-characterized oviduct epithelial cell line as well as an optimized transfection protocol for DNA vectors and siRNA for this cell line has been established. A bovine oviduct primary cell culture system has been optimized using a selection medium permitting the survival of only epithelial cells. From this we established an adherent bovine oviduct pure epithelial cell line (aBOPEC-1). This cell line maintains some important characteristics of the primary cells such as the expression of estrogen receptors and p450 aromatase but it lacks some characteristics due to the selection and dedifferentiation processes (cilia, expression of progesterone receptor and oviduct specific glycoprotein-1). Optimization of the transfection protocols finally revealed a suitable DNA-transfection procedure yielding transfection efficiencies of over 50%. Additionally, siRNA transfection efficiency reached more than 90%. This new cell line builds an essential basis especially for future functional studies in the oviduct epithelium using distinct knock down experiments.
Schoen
J
J
Institute of Veterinary Biochemistry, Freie Universität Berlin, Oertzenweg 19b, 14163 Berlin, Germany. schoen.jennifer@vetmed.fu-berlin.de
Bondzio
A
A
Topp
K
K
Einspanier
R
R
D000818:Animals; D002417:Cattle / Q000502:physiology*; D018929:Cell Culture Techniques / Q000662:veterinary; D002460:Cell Line; D002470:Cell Survival / Q000502:physiology; D004847:Epithelial Cells / Q000166:cytology* / Q000502:physiology; D047628:Estrogen Receptor alpha / Q000737:chemistry / Q000235:genetics; D047629:Estrogen Receptor beta / Q000737:chemistry / Q000235:genetics; D005187:Fallopian Tubes / Q000166:cytology* / Q000502:physiology; D005260:Female; D006023:Glycoproteins / Q000737:chemistry / Q000235:genetics; D007150:Immunohistochemistry / Q000662:veterinary; D012337:RNA, Ribosomal, 18S / Q000737:chemistry / Q000235:genetics; D034741:RNA, Small Interfering / Q000494:pharmacology; D011980:Receptors, Progesterone / Q000737:chemistry / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000662:veterinary; D014162:Transfection / Q000379:methods / Q000662:veterinary; D014170:Transformation, Genetic / Q000502:physiology
D047628:Estrogen Receptor alpha; D047629:Estrogen Receptor beta; D006023:Glycoproteins; C087523:OVGP1 protein, Bos taurus; D012337:RNA, Ribosomal, 18S; D034741:RNA, Small Interfering; D011980:Receptors, Progesterone