Efficient production of trophoblast lineage cells from human induced pluripotent stem cells

Abstract

Human induced pluripotent stem cells (hiPSCs) are potentially useful in both clinical applications and basic biological research. hiPSCs can differentiate into extra-embryonic cells in the presence of BMP4. However, the differentiation potential of hiPSCs can be affected by culture conditions or genetic variation. In this study, we investigated the effect of various BMP4 concentrations on the expression states of trophoblast markers and the optimal conditions for trophoblast induction. A high-fidelity gene expression assay using hiPSC lines showed that the expression levels of various trophoblast marker genes, such as KRT7, GCM1, CGB, and HLA-G, were upregulated by BMP4 in a dose-dependent manner in all types of hiPSCs used in this study. Treatment with high doses of BMP4 for prolonged periods increased the ratio of cells with trophoblast markers irrespective of the presence of bFGF. We found that the expression states of major pluripotency- and differentiation-related protein-coding genes in BMP4-treated cells depended on culture conditions rather than donor cell types. However, miRNA expression states were affected by donor cell types rather than BMP4 dose. Furthermore, the effect of the presence of bFGF on differentiation potential of KRT7-positive cells differed among iPSC types. Mechanistically, chromatin states around KRT7 promoter regions were comparable among the iPSC types used in this study, indicating that hiPSC chromatin state at these regions is not a parameter for cytotrophoblast differentiation potential. In conclusion, the optimal conditions for trophoblast differentiation from hiPSCs differ according to parental cell line.

Autonomous trisomic rescue of Down syndrome cells

“…Immortality of induced pluripotent stem cells (iPSCs) makes it possible to obtain a large number of cells from a small specimen, and pluripotency enables differentiation into various cell types [21–24]. Therefore, they are widely used to clarify disease etiology and test therapeutic drugs [25–28].…”

Toward a Reconceptualization of Stem Cells from Cellular Plasticity

“…In recent years, this strategy has been successfully applied in the transformation of fibroblasts into hepatocytes, nerve cells and cardiomyocytes. Third, somatic cells can also dedifferentiate into induced pluripotent stem cells that can further differentiate into any target cells even including trophoblast lineage cells (44–46).…”

The quest for pluripotency: a comparative analysis across mammalian species

“…Under in vitro conditions, pluripotent stem cells (PSCs) can beget all the cell types of embryonic and extra-embryonic lineages (reviewed in Beddington & Robertson 1989, Xu et al 2002, Pera et al 2004, Das et al 2007, Niakan et al 2010, Sudheer et al 2012, Leitch & Smith 2013, Morgani et al 2013, Roberts et al 2014, Kojima et al 2017. The potency of PSCs in giving rise to cells of the extra-embryonic lineage was first reported in mice, this was based on the presence of colonisation pattern of ESCs in trophectoderm (TE) of chimeras (Beddington & Robertson 1989).…”

“…The potency of PSCs in giving rise to cells of the extra-embryonic lineage was first reported in mice, this was based on the presence of colonisation pattern of ESCs in trophectoderm (TE) of chimeras (Beddington & Robertson 1989). Later on, the competency of hPSCs to differentiate into extraembryonic cells, such as trophoblast and primitive endoderm when exposed to BMP4 was described (Xu et al 2002, Pera et al 2004, Das et al 2007, Sudheer et al 2012, Horii et al 2016, Kojima et al 2017. In spite of several publications from different groups on the differentiation ability of human PSCs to extra-embryonic lineages, these protocols and concept are still debated among scientists in the field (Roberts et al 2014).…”