Isolation and Characterization of a Bovine Blastocyst-Derived Trophoblastic Cell line, BT-1: Development of a Culture System in the Absence of Feeder Cell

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

doi:10.1053/plac.2001.0702

Cited by 83 publications

(92 citation statements)

References 29 publications

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“…MNC production of IFNT is well known as a factor that signals the recognition of pregnancy; BNCs may play a major role in the establishment of the fetal-maternal interface, namely, BNC fuses with an epithelial cell in endometrium and starts to raise placental villi at around day 20 of gestation (Wooding 1992, Yamada et al 2002. Although cell lines are a convenient tool to analyze and understand the functions and the mechanisms of trophoblastic cell lineages, few bovine trophoblastic cell lines have been available (Talbot et al 2000, Shimada et al 2001. These facts encourage the establishment of new trophoblast cell lines.…”

Section: Discussionmentioning

confidence: 99%

“…Various cytokines and transcription factors may be involved; some previous BMP4 stimulates trophoblast cell lineage reports suggested that the activin/nodal pathway, BMPs, FGF4, FGF2, CDX, and POU5F1 (OCT4) are involved (Pfarrer et al 2006, Ravelich et al 2006, Sugawara et al 2010. In a previous study, we used only conditioned medium from fetal fibroblast cells and succeeded in producing only one cell line from about 50 IVF embryos, which has maintained good growth for over 300 passages (PDL; Shimada et al 2001, Hashizume et al 2006. The efficiency was quite low and it generally seems to be difficult to raise trophoblastic cell lines in cattle.…”

Section: Discussionmentioning

confidence: 99%

“…However, there are few available bovine trophoblastic cell lines. Various new kinds of cell lines will help in improving the understanding of the functions of trophoblastic cell lineages (Talbot et al 2000, Shimada et al 2001, Michael et al 2006, Hambruch et al 2010. In cattle, trophoblastic cells may have the potential to induce differentiation for different functional features of cells.…”

Section: Introductionmentioning

confidence: 99%

“…These facts suggest that the establishment of new trophoblast cell lines provide clue as a necessary tools for studying trophoblast cell lineage. Although we have been developing trophoblastic cell lines from early bovine embryos, the efficiency of generation of these cell lines has been limited (Shimada et al 2001).…”

Section: Introductionmentioning

confidence: 99%

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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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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

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

Suzuki¹,

Koshi²,

Imai³

et al. 2011

Reproduction

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“…This type of supplementation is not needed to maintain bovine trophectoderm in culture. They continue to proliferate without differentiating for extensive periods in culture (Talbot et al 2000, Shimada et al 2001.…”

Section: Introductionmentioning

confidence: 99%

Several fibroblast growth factors are expressed during pre-attachment bovine conceptus development and regulate interferon-tau expression from trophectoderm

Cooke¹,

Pennington²,

Yang³

et al. 2009

Reproduction

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The trophectoderm-derived factor interferon tau (IFNT) maintains the uterus in a pregnancy-receptive state in cattle and sheep. Fibroblast growth factors (FGFs) are implicated in regulating IFNT expression and potentially other critical events associated with early conceptus development in cattle. The overall objectives of this work were to identify the various FGFs and FGF receptors (FGFRs) expressed in elongating pre-attachment bovine conceptuses and determine if these FGFs regulate conceptus development and/or mediate IFNT production. In vitro-derived bovine blastocysts and in vivo-derived elongated conceptuses collected at day 17 of pregnancy express at least four FGFR subtypes (R1c, R2b, R3c, R4). In addition, transcripts for FGF1, 2, and 10 but not FGF7 are present in elongated bovine conceptuses. The expression pattern of FGF10 most closely resembled that of IFNT, with both transcripts remaining low in day 8 and day 11 conceptuses and increasing substantially in day 14 and day 17 conceptuses. Supplementation with recombinant FGF1, 2 or 10 increased IFNT mRNA levels in bovine trophectoderm cells and bovine blastocysts and increased IFNT protein concentrations in trophectoderm-conditioned medium. Blastocyst development was not affected by any of the FGFs. In summary, at least four FGFRs reside in pre-and peri-attachment bovine conceptuses. Moreover, conceptuses express at least three candidate FGFs during elongation, the time of peak IFNT expression. These findings provide new insight for how conceptus-derived factors such as FGF1, 2, and 10 may control IFNT expression during early pregnancy in cattle.

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Isolation and characterization of a bovine trophectoderm cell line derived from a parthenogenetic blastocyst

Talbot

Caperna

Powell

et al. 2004

Molecular Reproduction Devel

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A bovine trophectoderm cell line was established from a parthenogenetic in vitro-produced blastocyst. To initiate the cell line, 8-day parthenogenetic blastocysts were attached to a feeder layer of STO fibroblasts and primary outgrowths occurred that consisted of trophectoderm, endoderm, and very occasionally epiblast tissue. Any endoderm and epiblast outgrowths were removed from the primary cultures within the first 10 days of culture by dissection. One of the primary trophectoderm cell cultures was chosen for further propagation and was passaged by physical dissociation and replating on STO feeder cells. The cell culture, designated BPT-1, was maintained in T25 flasks and passaged at a 1:3 split ratio for the first 15 passages approximately once every 2 weeks. Thereafter, the cell culture was passaged at 1:10-1:40 split ratios. Transmission electron microscopic examination showed the cells to be a polarized epithelium with apical microvilli, a thin basal lamina, and lateral junctions consisting of tight junctions and desmosomes. Lipid vacuoles and digestive vacuoles were also prominent features of the BPT-1 cells. Metaphase spread analysis at passage 59 indicated a near diploid cell population (2n = 60) with a mode and median of 60 and a mean of 64. BPT-1 cells secreted interferon-tau into the medium as measured by anti-viral assay and Western blot analysis. The cell line provides an in vitro model of parthenogenote trophectoderm whose biological characteristics can be compared to trophectoderm cell lines derived from bovine embryos produced by normal fertilization or nuclear transfer.

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Isolation and Characterization of a Bovine Blastocyst-Derived Trophoblastic Cell line, BT-1: Development of a Culture System in the Absence of Feeder Cell

Cited by 83 publications

References 29 publications

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

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

Several fibroblast growth factors are expressed during pre-attachment bovine conceptus development and regulate interferon-tau expression from trophectoderm

Isolation and characterization of a bovine trophectoderm cell line derived from a parthenogenetic blastocyst

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