BMP4 induction of trophoblast from mouse embryonic stem cells in defined culture conditions on laminin

Abstract

Because mouse embryonic stem cells (mESCs) do not contribute to the formation of extraembryonic placenta when they are injected into blastocysts, it is believed that mESCs do not differentiate into trophoblast whereas human embryonic stem cells (hESCs) can express trophoblast markers when exposed to bone morphogenetic protein 4 (BMP4) in vitro. To test whether mESCs have the potential to differentiate into trophoblast, we assessed the effect of BMP4 on mESCs in a defined monolayer culture condition. The expression of trophoblast-specific transcription factors such as Cdx2, Dlx3, Esx1, Gata3, Hand1, Mash2, and Plx1 was specifically upregulated in the BMP4-treated differentiated cells, and these cells expressed trophoblast markers. These results suggest that BMP4 treatment in defined culture conditions enabled mESCs to differentiate into trophoblast. This differentiation was inhibited by serum or leukemia inhibitory factor, which are generally used for mESC culture. In addition, we studied the mechanism underlying BMP4-directed mESC differentiation into trophoblast. Our results showed that BMP4 activates the Smad pathway in mESCs inducing Cdx2 expression, which plays a crucial role in trophoblast differentiation, through the binding of Smad protein to the Cdx2 genomic enhancer sequence. Our findings imply that there is a common molecular mechanism underlying hESC and mESC differentiation into trophoblast.

Nodal Signaling Regulates the Bone Morphogenic Protein Pluripotency Pathway in Mouse Embryonic Stem Cells

“…More work needs to be done to determine if this derepression of trophectoderm genes is BMP-dependent. Mouse ES cells do not readily contribute to the trophectoderm lineages when introduced to a blastocyst; however, some differentiation conditions have been described to allow such differentiation in vitro (54,55). Thus, further work is needed to ascertain whether autocrine Nodal signaling inhibits the propensity of ES cells to differentiate to a trophectoderm pathway, and if this function is achieved by attenuating BMP activity.…”

Both BMP4 and serum have significant roles in differentiation of embryonic stem cells to primitive and definitive endoderm

“…In several studies, the effect of BMP4 on ES cell differentiation to endodermal lineages has been investigated in combination with the other factors. Moreover, some investigators have claimed that BMP4 alone can induce differentiation of human and mouse ES cells to trophoblastic lineages (Amita et al 2013;Hayashi et al 2010;Xu et al 2002). In the present study, we investigated the role of BMP4 signaling in differentiation of mouse ES cells to primitive and definitive endoderm.…”

Self-Renewal, Pluripotency and Tumorigenesis in Pluripotent Stem Cells Revisited

“…Interestingly, other extracellular matrix proteins elicit different responses from ESCs. For example, collagen IA promotes the self-renewal of mouse ESCs (Furue et al, 2005), and fibronectin and laminin help decrease their differentiation potential (Hayashi et al, 2007;Hayashi et al, 2010). Collagen IV is an inducer of mesoderm lineages for both mouse and human ESCs (SchenkeLayland et al, 2007).…”