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Establishment and characterization of Bovine trophoblast stem cells (bTSCs) growing as adherent cells or vesicles

ABSTRACT: Establishment and characterization of Bovine trophoblast stem cells (bTSCs) growing as adherent cells or vesicles

PROVIDER: PRJNA1226208 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR32415963.fastq.gz	Fastqsanger.gz
	SRR32415964.fastq.gz	Fastqsanger.gz
	SRR32415965.fastq.gz	Fastqsanger.gz
	SRR32415966.fastq.gz	Fastqsanger.gz
	SRR32415967.fastq.gz	Fastqsanger.gz

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Establishment of bovine trophoblast stem cells

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Establishment and characterization of Bovine trophoblast stem cells (bTSCs) growing as adherent cells or vesicles

Project description:Understanding embryonic development still remains one of the main challenge in developmental biology. Using 3D in vitro models of early embryonic development in the mouse and human, scientists hope to decipher embryonic mechanisms, in particular the initial arrangement and interaction of cells within the embryonic structure. To date, there are very few models established for livestock species. In this study, we have derived and characterized bovine trophoblastic stem cells (bTSCs) that we further used to establish with bESCs (bovine embryonic stem cells) the first in vitro model of bovine blastoïds.

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SECRETORY PROTEOMIC RESPONSES OF ENDOMETRIAL EPITHELIAL CELLS TO TROPHOBLAST DERIVED EXTRACELLULAR VESICLES

Project description:Synchronized cross talk between embryo and endometrium during pre-implantation period is critical for establishment of pregnancy. Extracellular vesicels(EVs) of both embryo and endometrial origin are known to be integral in regulating embryo maternal communication during this time. However, exact molecular signalling pathways or factors that regulate the embryo endometrial communication during pre-conception period remains elusive. Here in this study extracellular vesicles from trophoblast cells were shown to modulate endometrial epithelial cell secretome in favour of embryo implantation and embryo development.

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Establishment of trophoblast stem cells under defined culture conditions in mice

Project description:The inner cell mass (ICM) and trophoblast cell lineages duet early embryonic development in mammals. After implantation, the ICM forms the embryo proper as well as some extraembryonic tissues, whereas the trophoectoderm (TE) exclusively forms the fetal portion of the placenta and the trophoblast giant cells. Although embryonic stem (ES) cells can be derived from ICM in cultures of mouse blastocysts in the presence of LIF and/or combinations of small-molecule chemical compounds, and the undifferentiated pluripotent state can be stably maintained without use of serum and feeder cells, defined culture conditions for derivation and maintenance of undifferentiated trophoblast stem (TS) cells have not been established. Here, we report that addition of FGF2, activin A, XAV939, and Y27632 are necessary and sufficient for derivation of TS cells from both of E3.5 blastocysts and E6.5 early postimplantation extraembryonic ectoderm. Moreover, the undifferentiated TS cell state can be stably maintained in chemically defined culture conditions. Cells derived in this manner expressed TS cell marker genes, including Eomes, Elf5, Cdx2, Klf5, Cdh1, Esrrb, Sox2, and Tcfap2c; differentiated into all trophoblast subtypes (trophoblast giant cells, spongiotrophoblast, and labyrinthine trophoblast) in vitro; and exclusively contributed to trophoblast lineages in chimeric animals. This delineation of minimal requirements for derivation and self-renewal provides a defined platform for precise description and dissection of the molecular state of TS cells. Total RNA was purified by using RNeasy Mini kit (QIAGEN). Microarray targets from 200 ng total RNA were synthesized and labelled using the Low RNA Input Linear Amp Kit (Agilent) and hybridized to Mouse 4x44K Ver.2.0 arrays. Arrays were scanned on an Agilent Technologies Microarray scanner and signal intensities were calculated in Agilent Feature Extraction 10.7.3.1 software.

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