title: Identification of pluripotent cells in bovine uterus: in situ and in vitro studies.
abstract: The aim of this study was to identify uterine pluripotent cells both in bovine uterine tissues as well in epithelial, stromal, and myometrial uterine cell populations. Moreover, the relationship of pluripotent markers expression with age and the uterine horn side was considered. Uterine tissue was collected from ipsilateral and contralateral horns (days 8-10 of the estrous cycle). Immunohistostaining for C-KIT, OCT3/4, NANOG, and SOX2 in uterine tissue was determined. mRNA expression of C-KIT, OCT3/4, NANOG and SOX2 was evaluated in uterine tissue relative to the age of the cow and uterine horn side. Gene and protein expression of these markers in the uterine luminal epithelial, stromal, and myometrial cells was evaluated by real-time PCR and western blotting respectively. The expression of pluripotent cell markers OCT3/4, NANOG, and SOX2 was identified by flow cytometry assay in epithelial, stromal, and myometrial cells. Multilineage differentiation of the bovine uterine cells was performed. mRNA expression of OCT3/4, NANOG, and SOX2 in uterine tissue was higher in the ipsilateral horn than in the contralateral horn. Flow cytometry assay revealed positive fluorescence for OCT3/4, NANOG, and SOX2 in all uterine cell types. Results showed the age-dependent expression of pluripotent markers in uterine tissue. Beside, the different expression of pluripotent cells in each horn of uterus suggests the influence of ovarian hormones on these characteristics. The highest mRNA and protein expression for pluripotent markers was observed in stromal cells among uterine cells, which indicates this population of cells as the main site of pluripotent cells in the cow uterus.
authors.affiliation: Department of Reproductive Immunology and Pathology In Vitro and Cell Biotechnology Laboratory Institute of Animal Reproduction and Food Research, Polish Academy of Sciences, 10-748 Olsztyn, Poland Koret School of Veterinary Medicine The University of Jerusalem, Rehovot 76100, Israel.
authors.forename: Martyna
authors.initials: M
authors.lastname: Łupicka
authors.affiliation: Department of Reproductive Immunology and Pathology In Vitro and Cell Biotechnology Laboratory Institute of Animal Reproduction and Food Research, Polish Academy of Sciences, 10-748 Olsztyn, Poland Koret School of Veterinary Medicine The University of Jerusalem, Rehovot 76100, Israel.
authors.forename: Gabriel
authors.initials: G
authors.lastname: Bodek
authors.affiliation: Department of Reproductive Immunology and Pathology In Vitro and Cell Biotechnology Laboratory Institute of Animal Reproduction and Food Research, Polish Academy of Sciences, 10-748 Olsztyn, Poland Koret School of Veterinary Medicine The University of Jerusalem, Rehovot 76100, Israel.
authors.forename: Nahum
authors.initials: N
authors.lastname: Shpigel
authors.affiliation: Department of Reproductive Immunology and Pathology In Vitro and Cell Biotechnology Laboratory Institute of Animal Reproduction and Food Research, Polish Academy of Sciences, 10-748 Olsztyn, Poland Koret School of Veterinary Medicine The University of Jerusalem, Rehovot 76100, Israel.
authors.forename: Ehud
authors.initials: E
authors.lastname: Elnekave
authors.affiliation: Department of Reproductive Immunology and Pathology In Vitro and Cell Biotechnology Laboratory Institute of Animal Reproduction and Food Research, Polish Academy of Sciences, 10-748 Olsztyn, Poland Koret School of Veterinary Medicine The University of Jerusalem, Rehovot 76100, Israel a.korzekwa@pan.olsztyn.pl.
authors.forename: Anna J
authors.initials: AJ
authors.lastname: Korzekwa
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000378:metabolism*; D015153:Blotting, Western; D002417:Cattle; D002454:Cell Differentiation; D019070:Cell Lineage; D002478:Cells, Cultured; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D018507:Gene Expression Regulation, Developmental; D007124:Immunoenzyme Techniques; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014599:Uterus / Q000166:cytology* / Q000378:metabolism
chemical_list: D015415:Biomarkers; D012333:RNA, Messenger