Trophoblast differentiation

Series:	IN_VITRO DIFFERENTIATION SERIES
Species:	Mouse (Mus musculus)
Genomic View:	Zenbu
Expression table:	FILE
Link to TET:	TET
Sample providers :	Haruhiko Koseki
Germ layer:	ectoderm/ throphoblast
Primary cells or cell line:	cell line
Time span:	6 days
Number of time points:	7

Overview

During mouse development, the implanting blastocyst embryo is composed of the inner cell mass (ICM) and the outer trophectoderm (TE) layer. Permanent stem cell lines can be derived from these two lineages; embryonic stem (ES) cells from ICM [1] [2] and trophoblast stem (TS) cells from TE [3]. TS cells can differentiate into multiple cell types only of the trophoblast lineage, while ES cells can differentiated into all three germ layer cell types. TS cells grow in the presence of fibroblast growth factor (FGF) 4 and mouse embryonic fibroblast-conditioned medium (MEF-CM). Removal of FGF4 and MEF-CM from the TS cell culture medium causes them to differentiate in vitro into trophoblast giant cells and other trophoblast subtypes. We want to investigate how gene expression is controlled in the trophoblast lineage. To do this we conducted a timecourse analysis of gene expression profiles of TS cells after the removal of FGF4 and MEF-CM. References: [1] Evans MJ and Kaufman MH. Establishment in culture of pluripotential cells from mouse embryos. Nature. 1981. 292(5819): p154-6. [2] Martin GR. Martin, Proc Natl Acad Sci U S A. 1981. 78(12): p7634-8. [3] Tanaka S. Promotion of trophoblast stem cell proliferation by FGF4. Science. 1998. 282(5396): p2072-5.

Sample description

TS cell lines are maintained in an undifferentiated state in the presence of FGF4 and 70% MEF-CM, and are differentiated by removal of FGF4 and MEF-CM for 0, 1, 2, 3, 4, 5, or 6 days (total 7 points). We used following three TS cell lines: a wild-type female TS cell line “B1”, a wild-type male TS cell line “Rybp”, and an Ring1A-/- male TS cell line “R1AB” (Ring1A-/- mice are healthy and exhibit no apparent phenotype in placental tissues [4]) . References: [4] del Mar Lorente M, Marcos-Gutiérrez C, Pérez C, Schoorlemmer J, Ramírez A, Magin T, Vidal M. Loss- and gain-of-function mutations show a polycomb group function for Ring1A in mice. Development. 2000. 127(23): p5093-100.

Quality control

List of published key marker genes for your time course, if available with references: * Cdx2, Eomes, Esrrb, Fgfr2: Marker genes for undifferentiated TS cells [3] * Prl3d1, Prl3b1, Prl2c2: Marker genes for trophoblast giant cells [5] [6] * Tpbpa: The Tpbpa gene is expressed in ectoplacental cone cells starting between embryonic days (E) 7.5 and 8.5, and later in the spongiotrophoblast layer of the mature placenta [7] * Ascl2: The Ascl2 gene is expressed in the ectoplacental cone, the chorion and their derivatives in the placenta [8] Figure 1: B1 TS cells day0 (undifferentiated; scale bar = 100 um) Figure 2: B1 TS cells day5-differentiation (scale bar = 100 um) Figure 3: R1AB TS cells day0 (undifferentiated; scale bar = 100 um) Figure 4: R1AB TS cells day5-differentiation (scale bar = 100 um) Figure 5: CAGE expression of marker genes in TPM. References: [3] Tanaka S. Promotion of trophoblast stem cell proliferation by FGF4. Science. 1998. 282(5396): p2072-5. [5] Faria TN, Ogren L, Talamantes F, Linzer DI, Soares MJ. Localization of placental lactogen-I in trophoblast giant cells of the mouse placenta. Biol Reprod. 1991. 44(2), p327-31 [6] Hu D and Cross JC. Development and function of trophoblast giant cells in the rodent placenta. Int J Dev Biol. 2010. 54(2-3): p341-54 [7] Hu D and Cross JC. Ablation of Tpbpa-positive trophoblast precursors leads to defects in maternal spiral artery remodeling in the mouse placenta. Dev Biol. 2011. 358(1): p231-9 [8] Guillemot F, Nagy A, Auerbach A, Rossant J, Joyner AL. Essential role of Mash-2 in extraembryonic development. Nature 1994. 371 (6495): p.333-6

Profiled time course samples

Only samples that passed quality controls (Arner et al. 2015) are shown here. The entire set of samples are downloadable from FANTOM5 human / mouse samples

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13283-142F7	trophoblast stem cell line R1AB differentiation	day00	biol_rep1
13284-142F8	trophoblast stem cell line R1AB differentiation	day01	biol_rep1
13285-142F9	trophoblast stem cell line R1AB differentiation	day02	biol_rep1
13286-142G1	trophoblast stem cell line R1AB differentiation	day03	biol_rep1
13287-142G2	trophoblast stem cell line R1AB differentiation	day04	biol_rep1
13288-142G3	trophoblast stem cell line R1AB differentiation	day05	biol_rep1
13289-142G4	trophoblast stem cell line R1AB differentiation	day06	biol_rep1
13290-142G5	trophoblast stem cell line Rybp differentiation	day00	biol_rep1
13291-142G6	trophoblast stem cell line Rybp differentiation	day01	biol_rep1
13292-142G7	trophoblast stem cell line Rybp differentiation	day02	biol_rep1
13293-142G8	trophoblast stem cell line Rybp differentiation	day03	biol_rep1
13294-142G9	trophoblast stem cell line Rybp differentiation	day04	biol_rep1
13295-142H1	trophoblast stem cell line Rybp differentiation	day05	biol_rep1
13296-142H2	trophoblast stem cell line Rybp differentiation	day06	biol_rep1
13297-142H3	trophoblast stem cell line B1 differentiation	day00	biol_rep1
13298-142H4	trophoblast stem cell line B1 differentiation	day01	biol_rep1
13299-142H5	trophoblast stem cell line B1 differentiation	day02	biol_rep1
13300-142H6	trophoblast stem cell line B1 differentiation	day03	biol_rep1
13301-142H7	trophoblast stem cell line B1 differentiation	day04	biol_rep1
13302-142H8	trophoblast stem cell line B1 differentiation	day05	biol_rep1
13303-142H9	trophoblast stem cell line B1 differentiation	day06	biol_rep1