Bovine trophoblastic cell differentiation on collagen substrata: formation of binucleate cells expressing placental lactogen

Nakano, Hitoo; Shimada, Arata; Imai, Kyoichi; Takezawa, Toshiaki; Takahashi, Toshio; Hashizume, Kazuyoshi

doi:10.1007/s00441-001-0491-x

Cited by 43 publications

(34 citation statements)

References 41 publications

(74 reference statements)

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“…This morphological appearance suggests that the term synepitheliochorial placenta is a more suitable appellation for ruminant placenta. The epithelium may be reestablished after placentation [36]. At the same time, BNCs expressed PRP-1 and heparanase in limited areas [33,37].…”

Section: Role Of Mmps In Endometrial Remodeling Around the Time Of Immentioning

confidence: 99%

Analysis of Uteroplacental-specific Molecules and Their Functions During Implantation and Placentation in the Bovine

Hashizume

2007

Journal of Reproduction and Development

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Abstract. In cattle, the mechanisms underlying implantation and placental development are still unclear. Synepitheliochorial placentation in cattle is noninvasive, and thus generates limited interest in terms of degradation and remodeling of endometrial tissues. The overall purpose of this study was three-fold: (1) to examine the gene circuitry around the implantation window, (2) to understand development of the placenta during the peri-implantation period by using a uteroplacental cDNA microarray, and (3) to study the roles of molecules involved in endometrial remodeling. Bovine trophoblastic binucleate cell-specific molecules, such as pregnancy-associated glycoproteins (PAGs), placental lactogen (PL), and prolactin-related proteins (PRPs), were markedly expressed in binucleate cells (BNCs) around implantation. The expression of PRP-1 was specific to the caruncular (CAR) area of the gravid uterine horn. Gelatinases (MMP-2 and -9) in association with heparanase may be central to endometrial remodeling. In situ hybridization analyses of PAGs, PRPs, PL, and heparanase suggested that BNCs expressed these molecules simultaneously. Future studies will further investigate the specific roles of these molecules in placentogenesis. The uteroplacental cDNA microarray presented cascades of molecular signatures not only for the endometrium but also for the intricate dialogue at the level of the feto-maternal interface in cattle. Placentome morphogenesis potentially parallels the dynamic multigenic circuitry and regulates the cell cycle in the endometrium. The roles of BNCs and their secreted molecules remain an enigma, particularly with regard to the adhesion process and endometrial remodeling, which is the focus of this study.

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Section: Role Of Mmps In Endometrial Remodeling Around the Time Of Immentioning

confidence: 99%

Analysis of Uteroplacental-specific Molecules and Their Functions During Implantation and Placentation in the Bovine

Hashizume

2007

Journal of Reproduction and Development

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“…After separation, proteins were transferred onto PVDF membrane (Immobilon-P, Millipore, Billerica, MA, USA) with semi-dry blotter (Bio-Rad Japan). The transferred membranes were detected with anti-IFNT (!2000) antisera as described in previous reports (Shimada et al 2001, Nakano et al 2002.…”

Section: Western Blottingmentioning

confidence: 99%

“…In ruminants, a mononucleate trophoblast cell (MNC) may directly differentiate into a binucleate cell (BNC) without cytokinesis and the BNC then fuses with an endometrial epithelial cell to form a trinucleate cell (Nakano et al 2002, Fléchon et al 2007, Ullah et al 2009). This event is the start of implantation in ruminants; during the peri-implantation period, the MNC and BNC (or multinucleate) cells produce various molecules for placentogenesis (Spencer et al 2004).…”

Section: Introductionmentioning

confidence: 99%

Bone morphogenetic protein 4 accelerates the establishment of bovine trophoblastic cell lines

Suzuki¹,

Koshi²,

Imai³

et al. 2011

Reproduction

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Trophoblastic cells play a crucial role in implantation and placentogenesis. A large proportion of the failures of conception in cows occur in the peri-implantation period, which are known as early embryo losses. In exploring this critical phenomenon, trophoblastic cell lines can provide substantial information. Unfortunately, there are few cell lines for this purpose in cattle because of the difficulty of raising successive cell stock in the long term. In this study, 12 new cell lines were established using bone morphogenetic protein 4 (BMP4). BMP4 stimulated embryonic cells to enter the trophoblastic cell lineage but there were no significant differences between intact and BMP4-treated groups. Only one out of 49 embryos developed trophoblastic cells in the intact group. Finally, 12 cell lines were maintained for around 30 passages, and they retained trophoblastic characteristics and expressed bovine trophoblastic genes: placental lactogen, interferon-t, pregnancy-associated glycoprotein 1, and prolactin-related protein 1. Although the gene expression patterns were different among cell lines and depended on the cells, there was no significant relationship between the expression intensities of genes and the treatment dose of BMP4. All of them expressed bovine POU domain class 5 transcription factor 1 and caudal-type homeobox 2. The expression of these genes was confirmed by quantitative RT-PCR and immunohistochemical detection. These results suggest that BMP4 is involved in the raising of trophoblast cell lines from early embryonic cells and the newly developed cell lines can provide different types of bovine trophoblastic cells with different cell lineages. This may constitute a significant new tool for the examination of trophoblastic differentiation.

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“…BT-1 established from in vitro blastocysts [11,12] was cultured in Dulbecco's modified Eagle's/F-12 medium (DME/F-12, GIBCO Invitrogen, Carlsbad, CA, USA) containing 100 IU/ml of penicillin and 100 µg/ml of streptomycin (Sigma, St. Louis, MO, USA), supplemented with 10% fetal bovine serum (FBS, Sigma), 50% bovine uterine fibroblast conditioned medium (UFCM) [11,12], 50 µmol/l β-mercaptoethanol, and 1 µg/ml heparin at 37 C in an atmosphere of 5% CO 2 . The medium was changed every two or three days.…”

Section: Culture Of Bt-1mentioning

confidence: 99%

“…The cell clumps attached to the flasks and proliferated to re-form a monolayer. Successful passages were also achieved b y t r a n s f e r r i n g m u l t i c e l l u l a r v e s i c l e s spontaneously formed from the cell colony [11,12].…”

Section: Culture Of Bt-1mentioning

confidence: 99%

Gene Expression Profiles of Bovine Trophoblastic Cell Line (BT-1) Analyzed by a Custom cDNA Microarray

Ushizawa

Takahashi

Kaneyama

et al. 2005

Journal of Reproduction and Development

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Abstract. Gene expression of bovine trophoblast cell line (BT-1) was analyzed with a custom uteroplacenta complementary DNA microarray. Expression comparison with in vivo tissues of trophoblast derivation was performed to investigate characteristics of the expression in BT-1. BT-1 is a cell line established without feeder cells using trophoblast cells that are separated from bovine blastocysts. The bovine in vivo tissues of the trophoblast derivation were collected on day 17 through 56 of gestation. Of 1,773 genes, 933 exhibited an expression difference exceeding two-fold between BT-1 and in vivo tissues. These genes were analyzed by the k-means clustering method and were distributed into six clusters. Some genes, such as placental lactogen, interferon-τ and pregnancyassociated glycoproteins, all known as trophoblast-specific genes, were detected in BT-1 as well as in vivo tissues throughout the experiment period. These trophoblast-specific genes and octamer-binding transcription factor-4, known as a marker for undifferentiation of cells in mice, were detected by RT-PCR in both BT-1 and in vivo trophoblast tissues. The overall gene expression profile in BT-1 suggests that this cell line contains trophoblast-specific characteristics and is similar to trophoblast cells around the implantation period.

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Bovine trophoblastic cell differentiation on collagen substrata: formation of binucleate cells expressing placental lactogen

Cited by 43 publications

References 41 publications

Analysis of Uteroplacental-specific Molecules and Their Functions During Implantation and Placentation in the Bovine

Analysis of Uteroplacental-specific Molecules and Their Functions During Implantation and Placentation in the Bovine

Bone morphogenetic protein 4 accelerates the establishment of bovine trophoblastic cell lines

Gene Expression Profiles of Bovine Trophoblastic Cell Line (BT-1) Analyzed by a Custom cDNA Microarray

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