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Generation of trophoblast stem cells from rabbit embryonic stem cells with BMP4.

Tan T, Tang X, Zhang J, Niu Y, Chen H, Li B, Wei Q, Ji W - PLoS ONE (2011)

Bottom Line: Trophoblast stem (TS) cells are ideal models to investigate trophectoderm differentiation and placental development.Rabbit ES cells generated in our laboratory were induced to differentiate in the presence of BMP4 and TS-like cell colonies were isolated and expanded.These cells expressed the molecular markers of mouse TS cells, were able to invade, give rise to derivatives of TS cells, and chimerize placental tissues when injected into blastocysts.

View Article: PubMed Central - PubMed

Affiliation: Kunming Primate Research Center, Kunming Institute of Zoology, Chinese Academy of Sciences, Kunming, Yunnan, China.

ABSTRACT
Trophoblast stem (TS) cells are ideal models to investigate trophectoderm differentiation and placental development. Herein, we describe the derivation of rabbit trophoblast stem cells from embryonic stem (ES) cells. Rabbit ES cells generated in our laboratory were induced to differentiate in the presence of BMP4 and TS-like cell colonies were isolated and expanded. These cells expressed the molecular markers of mouse TS cells, were able to invade, give rise to derivatives of TS cells, and chimerize placental tissues when injected into blastocysts. The rabbit TS-like cells maintained self-renewal in culture medium with serum but without growth factors or feeder cells, whilst their proliferation and identity were compromised by inhibitors of FGFs and TGF-β receptors. Taken together, our study demonstrated the derivation of rabbit TS cells and suggested the essential roles of FGF and TGF-β signalings in maintenance of rabbit TS cell self-renewal.

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Derivation of epithelial-like cells from rabbit ES cells.(A) Epithelial-like cell clone. (B) Fibroblast-like cell clone. (C) Multinucleated cells formed in monolayer culture. Arrow pointed to the cell nucleus; magnification 200×. (D) Giant cells spontiniously differentiated from epithelial-like cell culture. Arrow pointed to cell nucleus. All magnifications were 100×, unless otherwise stated. The scale bar represents 100 µm.
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pone-0017124-g001: Derivation of epithelial-like cells from rabbit ES cells.(A) Epithelial-like cell clone. (B) Fibroblast-like cell clone. (C) Multinucleated cells formed in monolayer culture. Arrow pointed to the cell nucleus; magnification 200×. (D) Giant cells spontiniously differentiated from epithelial-like cell culture. Arrow pointed to cell nucleus. All magnifications were 100×, unless otherwise stated. The scale bar represents 100 µm.

Mentions: Rabbit ES cells treated with BMP4 were induced to differentiate into epithelial-like cells in both EB and monolayer culture systems. In EB differentiation, cells displayed heterogeneity at the beginning of BMP4 treatment (day 0), with cuboidal epithelial-like cells surrounded by fibroblast-like cells at the edge. The epithelial-like cells proliferated faster than the fibroblast-like cells, leading to domination of the epithelial-like population at day 10–15 of differentiation. A few multinucleated cells were formed at this stage. There was no significant difference among the four groups of BMP4 treatment in term of differentiation rates (1, 5, 10 and 20 ng/ml). These epithelial-like (Figure 1A) and fibroblast-like cells (Figure 1B) were expanded via limited dilution and individual cell clones were established from single cells. The epithelial-like cells were capable of self-renewal and have been passaged up to 60 times. The doubling time was 16.084±0.379 hours.


Generation of trophoblast stem cells from rabbit embryonic stem cells with BMP4.

Tan T, Tang X, Zhang J, Niu Y, Chen H, Li B, Wei Q, Ji W - PLoS ONE (2011)

Derivation of epithelial-like cells from rabbit ES cells.(A) Epithelial-like cell clone. (B) Fibroblast-like cell clone. (C) Multinucleated cells formed in monolayer culture. Arrow pointed to the cell nucleus; magnification 200×. (D) Giant cells spontiniously differentiated from epithelial-like cell culture. Arrow pointed to cell nucleus. All magnifications were 100×, unless otherwise stated. The scale bar represents 100 µm.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3040765&req=5

pone-0017124-g001: Derivation of epithelial-like cells from rabbit ES cells.(A) Epithelial-like cell clone. (B) Fibroblast-like cell clone. (C) Multinucleated cells formed in monolayer culture. Arrow pointed to the cell nucleus; magnification 200×. (D) Giant cells spontiniously differentiated from epithelial-like cell culture. Arrow pointed to cell nucleus. All magnifications were 100×, unless otherwise stated. The scale bar represents 100 µm.
Mentions: Rabbit ES cells treated with BMP4 were induced to differentiate into epithelial-like cells in both EB and monolayer culture systems. In EB differentiation, cells displayed heterogeneity at the beginning of BMP4 treatment (day 0), with cuboidal epithelial-like cells surrounded by fibroblast-like cells at the edge. The epithelial-like cells proliferated faster than the fibroblast-like cells, leading to domination of the epithelial-like population at day 10–15 of differentiation. A few multinucleated cells were formed at this stage. There was no significant difference among the four groups of BMP4 treatment in term of differentiation rates (1, 5, 10 and 20 ng/ml). These epithelial-like (Figure 1A) and fibroblast-like cells (Figure 1B) were expanded via limited dilution and individual cell clones were established from single cells. The epithelial-like cells were capable of self-renewal and have been passaged up to 60 times. The doubling time was 16.084±0.379 hours.

Bottom Line: Trophoblast stem (TS) cells are ideal models to investigate trophectoderm differentiation and placental development.Rabbit ES cells generated in our laboratory were induced to differentiate in the presence of BMP4 and TS-like cell colonies were isolated and expanded.These cells expressed the molecular markers of mouse TS cells, were able to invade, give rise to derivatives of TS cells, and chimerize placental tissues when injected into blastocysts.

View Article: PubMed Central - PubMed

Affiliation: Kunming Primate Research Center, Kunming Institute of Zoology, Chinese Academy of Sciences, Kunming, Yunnan, China.

ABSTRACT
Trophoblast stem (TS) cells are ideal models to investigate trophectoderm differentiation and placental development. Herein, we describe the derivation of rabbit trophoblast stem cells from embryonic stem (ES) cells. Rabbit ES cells generated in our laboratory were induced to differentiate in the presence of BMP4 and TS-like cell colonies were isolated and expanded. These cells expressed the molecular markers of mouse TS cells, were able to invade, give rise to derivatives of TS cells, and chimerize placental tissues when injected into blastocysts. The rabbit TS-like cells maintained self-renewal in culture medium with serum but without growth factors or feeder cells, whilst their proliferation and identity were compromised by inhibitors of FGFs and TGF-β receptors. Taken together, our study demonstrated the derivation of rabbit TS cells and suggested the essential roles of FGF and TGF-β signalings in maintenance of rabbit TS cell self-renewal.

Show MeSH
Related in: MedlinePlus