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Embryonic Stem Cells |
1 Department of Medicine and Physiology, Cardiovascular Research Laboratory, University of California Los Angeles, Los Angeles, California
2 Department of Molecular, Cellular and Developmental Biology, University of California Los Angeles, Los Angeles, California
3 Department of Surgery, Regenerative Bioengineering and Repair Laboratory, University of California Los Angeles, Los Angeles, California
* To whom correspondence should be addressed. E-mail: rmaclellan{at}mednet.ucla.edu.
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Abstract |
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The earliest segregation of lineages in the developing embryo is the commitment of cells to the inner cell mass (ICM) or the trophoectoderm (TE) in pre-implantation blastocysts. The exogenous signals that control commitment to a particular cell lineage are poorly understood; however, it has been suggested that extracellular 'niche' and extracellular matrix (ECM) in particular, plays an important role in determining the developmental fate of stem cells. Collagen IV (ColIV) has been reported to direct ES cell differentiation to mesodermal lineages in both mouse and human ES cells. To define the effects of ColIV on ES cell differentiation and to identify the resulting heterogeneous cell types, we performed microarray analyses and determined global gene expression. We observed that ColIV induced the expression of mesodermal genes specific to hematopoietic, endothelial and smooth muscle cells, and surprisingly, also a panel of trophoectoderm-restricted markers. This effect was specific to collagen IV, as no trophoblast differentiation was seen on collagen I, laminin or fibronectin. Stimulation with basic fibroblast growth factor (bFGF) or fibroblast growth factor 4 (FGF4) increased the number of trophoectodermal cells. These cells were isolated under clonal conditions and successfully differentiated into a variety of trophoblast derivatives. Interestingly, differentiation of ES cells to trophoblastic lineages was only seen in ES cell lines maintained on embryonic feeder layers and was Cdx2-dependent, consistent with Cdx2's postulated role in trophoectoderm commitment. Our data suggest that given the appropriate extracellular stimuli, mouse embryonic stem cells can differentiate into trophoectoderm.
Key Words. collagen IV, embryonic stem cells, extracellular matrix, trophoblast, placenta
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