Development of an in vitro Model for Bovine Placentation: A Comparison of the in vivo and in vitro Expression of Integrins and Components of Extracellular Matrix in Bovine Placental Cells

Zeiler, M.; Leiser, Rudolf; Johnson, Greg A.; Tinneberg, H.-R.; Pfarrer, Christiane

doi:10.1159/000107947

Cited by 33 publications

(38 citation statements)

References 109 publications

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“…Similar to bovine imEECs, the bovine EECs established in this study expressed both an epithelial marker, cytokeratin, and a stromal marker, vimentin (Zhang et al 2013). The acquisition of vimentin by cytokeratin-positive epithelial cells during in vitro culture periods has been reported (Bergh et al 1984;Wang et al 2000;Zeiler et al 2007), which has been explained by a limited epithelial to mesenchymal transition (EMT; Pagan et al 1996). This transition is believed to correlate with a loss of cell-to-cell contacts that result from cell disaggregation during the period of epithelial cell culture preparation (Zeiler et al 2007).…”

Section: Discussionsupporting

confidence: 66%

“…The acquisition of vimentin by cytokeratin-positive epithelial cells during in vitro culture periods has been reported (Bergh et al 1984;Wang et al 2000;Zeiler et al 2007), which has been explained by a limited epithelial to mesenchymal transition (EMT; Pagan et al 1996). This transition is believed to correlate with a loss of cell-to-cell contacts that result from cell disaggregation during the period of epithelial cell culture preparation (Zeiler et al 2007). In our study, the vimentin expression may have been acquired during the long deoxyribonuclease I and collagenase I digestion.…”

Section: Discussionmentioning

confidence: 99%

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The bovine endometrial epithelial cells promote the differentiation of trophoblast stem-like cells to binucleate trophoblast cells

Hou

et al. 2016

Cytotechnology

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Endometrial epithelial cells (EECs) cultured in vitro are valuable tools for investigating embryo implantation and trophoblast differentiation. In this study, we have established the bovine EECs and trophoblast stem-like (TS) coculture system, and used it to investigate the binucleate cell formation of ungulates. The EECs was derived from the uterine horn ipsilateral to the corpus luteum by using collagenase I and deoxyribonuclease I, which exhibited typical epithelial morphology and were expressing bovine uterine epithelial marker such as IFNAR1, IFNAR2, Era, PGR, ESR1 and KRT18. The cells immunostained positively by epithelial and trophectoderm marker cytokeratin 18 (KRT18) and stromal marker vimentin antibodies, and the KRT18 positive cells reached 99 %. The EECs can be cultured for up to 20 passages in vitro with no significant morphology changes and uterine epithelial marker gene expression alteration. The bTS cells were established in a dual inhibitor system and exhibited typical trophoblast stem cell characteristics. When bTS cells were cultured with EECs, the bTS cells adhered to the EECs as adhering to feeder cells. Binucleate cells began appearing on day 4 of coculture and reached approximately 18.47 % of the differentiated cells. Quantitative real-time PCR or immunofluorescence analyses were performed on bTS cells cocultured at day 6 and day 12. The results showed that the expression level of KRT18 was down-regulated while the expression level of trophoblast differentiation marker MASH2, HAND1, GCM1 and CDX2 was upregulated in bTS cells. In conclusion, bovine EECs can be obtained from the uterine horn ipsilateral to the corpus luteum via treatment with collagenase I and deoxyribonuclease I, and the EECs-bTS cells coculture system presents an ideal tool for studying the differentiation of bTS cells to trophoblast binucleate cells.

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Section: Discussionsupporting

confidence: 66%

Section: Discussionmentioning

confidence: 99%

The bovine endometrial epithelial cells promote the differentiation of trophoblast stem-like cells to binucleate trophoblast cells

Hou

et al. 2016

Cytotechnology

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“…The medium was changed every 2 days until confluence was reached. Cell culture homogeneity was confirmed using real-time PCR for mRNA expression and determination of VE-cadherin, vimentin, and desmin in epithelial, stromal, and myometrial cells respectively (Zeiler et al 2007).…”

Section: Uterine Cells Isolation and In Vitro Culturesmentioning

confidence: 97%

Identification of pluripotent cells in bovine uterus: in situ and in vitro studies

Łupicka

Bodek

Shpigel³

et al. 2015

Reproduction

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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.

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“…In this study, cultured EECs maintained their characteristic intermediate filaments, in that epithelial cells expressed an epithelial cell marker, cytokeratin, and a mesenchymal cell marker, vimentin. Cytokeratin-positive epithelial cells frequently acquire vimentin during culture (Zeiler et al 2007. In monocultures, EECs placed onto collagen type IA-coated six-well dishes were incubated with 10 mg proteins from the day 22 bovine uterine flushing, epidermal growth factor (EGF) (1, 5, 10, 20, and 40 ng/ml), basic fibroblast growth factor (bFGF) (1, 2.5, 5, 10, and 20 ng/ml), or IFNT (0.01, 0.05, 0.1, 1, and 10 mg/ml) in serum-free DMEM/F12 for 48 h.…”

Section: Cell Preparation and Culture Conditionmentioning

confidence: 99%

Involvement of VCAM1 in the bovine conceptus adhesion to the uterine endometrium

Bai

Kuse

et al. 2014

Reproduction

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Following bidirectional communication, the conceptus and the uterine epithelium must establish a proper cell-cell interaction, resulting in the progression of implantation processes. To clarify the mechanism of conceptus attachment to the uterine endometrium, we studied whether vascular cell adhesion molecule (VCAM1) was expressed in bovine conceptuses or endometrium during the peri-attachment period. Uterine VCAM1 expression was minimal in day 17 (day 0Zday of estrus) cyclic and pregnant animals, but increased between days 20 and 22 of pregnancy. In the intercaruncular regions, VCAM1 protein was localized to the luminal and glandular epithelia, whereas in the caruncular regions, VCAM1 protein was detected in the stroma and endothelia of the uterine endometrium. In cultured endometrial epithelial cells (EECs), VCAM1 expression was up-regulated when treated with uterine flushings or growth factor and further increased when EECs were cocultured with bovine trophoblast CT1 cells. VCAM1 expression in CT1 cells was also up-regulated with the use of uterine flushings, and further increased when these cells were cocultured with EECs. Expression of VCAM1 receptor, integrin a 4 (ITGA4) mRNA, increased significantly in day 22 conceptuses. In day 22 pregnant uteri, VCAM1 protein was found in both EECs and conceptuses, but ITGA4 was localized only to trophoblasts. These observations indicate that cell-cell interactions between conceptuses and uterine epithelial cells are required for sufficient VCAM1 and ITGA4 expression in the bovine species and suggest that uterine VCAM1 and conceptus ITGA4 play a role in the establishment of conceptus adhesion to the uterine endometrium.

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Development of an in vitro Model for Bovine Placentation: A Comparison of the in vivo and in vitro Expression of Integrins and Components of Extracellular Matrix in Bovine Placental Cells

Cited by 33 publications

References 109 publications

The bovine endometrial epithelial cells promote the differentiation of trophoblast stem-like cells to binucleate trophoblast cells

The bovine endometrial epithelial cells promote the differentiation of trophoblast stem-like cells to binucleate trophoblast cells

Identification of pluripotent cells in bovine uterus: in situ and in vitro studies

Involvement of VCAM1 in the bovine conceptus adhesion to the uterine endometrium

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