FGF4 Independent Derivation of Trophoblast Stem Cells from the Common Vole

Abstract

The derivation of stable multipotent trophoblast stem (TS) cell lines from preimplantation, and early postimplantation mouse embryos has been reported previously. FGF4, and its receptor FGFR2, have been identified as embryonic signaling factors responsible for the maintenance of the undifferentiated state of multipotent TS cells. Here we report the derivation of stable TS-like cell lines from the vole M. rossiaemeridionalis, in the absence of FGF4 and heparin. Vole TS-like cells are similar to murine TS cells with respect to their morphology, transcription factor gene expression and differentiation in vitro into derivatives of the trophectoderm lineage, and with respect to their ability to invade and erode host tissues, forming haemorrhagic tumours after subcutaneous injection into nude mice. Moreover, vole TS-like cells carry an inactive paternal X chromosome, indicating that they have undergone imprinted X inactivation, which is characteristic of the trophoblast lineage. Our results indicate that an alternative signaling pathway may be responsible for the establishment and stable proliferation of vole TS-like cells.

Trophoblast development

“…Recently, FGF-dependent rat TS cells, showing highly similar expression profiles to mouse TS cells, have been derived from blastocyst outgrowths (Asanoma et al 2011). However, TS lines that are independent of FGF signalling have also been established in rats as well as in another rodent, the common vole (Grigor'eva et al 2009, Chuykin et al 2010. A trophoblast cell line derived from rhesus monkey blastocysts was maintained independently of FGF signalling (Vandevoort et al 2007).…”

Difference between random and imprinted X inactivation in common voles

“…Cells were derived as previously described (Nesterova et al 1994) and maintained in Ham's F12 medium or in a mixture of Ham's F12 and D-MEM (1:1, GIBCO/BRL) supplemented with 10% fetal bovine serum, 1 mM L-glutamine (GIBCO/BRL), 50 U/ml penicillin (GIBCO/BRL), and 50 µg/ml streptomycin (GIBCO/BRL) at 37°С in an atmosphere of 5% CO 2 . Vole extraembryonic endoderm (XEN) and TS cells used for X-linked gene methylation analysis were maintained as described (Grigor'eva et al 2009;Shevchenko et al 2008Shevchenko et al , 2009). …”

“…3). In vole TS cells, XCI is imprinted as in XEN cells (Grigor'eva et al 2009). TS cells are derived from trophoblast, a major contributor to placenta, so one can extrapolate from TS cells phenomena occurring in the trophoblast lineage of extraembryonic tissues.…”

Dynamic Changes in Gene Expression during Early Trophoblast Differentiation from Human Embryonic Stem Cells Treated with BMP4

“…Such TSC can be cultured in a continuously self-renewing state in vitro in the presence of FGF4 plus heparin, but also require NODAL signaling, which is usually derived from a supporting feeder layer of primary embryonic fibroblasts (Tanaka, Kunath et al 1998;Rossant 2001;Rossant 2007). However, it should be noted that dependency on FGF4 may not be a universal feature of TSC from all species (Vandevoort, Thirkill et al 2007;Grigor'eva, Shevchenko et al 2009). There have been many excellent reviews on embryonic and trophoblast stem cells (TSC), and the differences that distinguish them.…”