title: 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.
authors.affiliation: Center for Regenerative Medicine, National Research Institute for Child Health and Development, Tokyo, Japan.|Department of Obstetrics and Gynecology, Tokyo Medical University, Tokyo, Japan.
authors.forename: Junya
authors.initials: J
authors.lastname: Kojima
authors.affiliation: Center for Regenerative Medicine, National Research Institute for Child Health and Development, Tokyo, Japan.
authors.forename: Atsushi
authors.initials: A
authors.lastname: Fukuda
authors.affiliation: Center for Regenerative Medicine, National Research Institute for Child Health and Development, Tokyo, Japan.
authors.forename: Hayato
authors.initials: H
authors.lastname: Taira
authors.affiliation: Center for Regenerative Medicine, National Research Institute for Child Health and Development, Tokyo, Japan.
authors.forename: Tomoyuki
authors.initials: T
authors.lastname: Kawasaki
authors.affiliation: Department of Obstetrics and Gynecology, Tokyo Medical University, Tokyo, Japan.
authors.forename: Hiroe
authors.initials: H
authors.lastname: Ito
authors.affiliation: Department of Obstetrics and Gynecology, Tokyo Medical University, Tokyo, Japan.
authors.forename: Naoaki
authors.initials: N
authors.lastname: Kuji
authors.affiliation: Department of Obstetrics and Gynecology, Tokyo Medical University, Tokyo, Japan.
authors.forename: Keiichi
authors.initials: K
authors.lastname: Isaka
authors.affiliation: Center for Regenerative Medicine, National Research Institute for Child Health and Development, Tokyo, Japan.
authors.forename: Akihiro
authors.initials: A
authors.lastname: Umezawa
authors.affiliation: Center for Regenerative Medicine, National Research Institute for Child Health and Development, Tokyo, Japan.|Department of Stem Cell Research, Fukushima Medical University, Fukushima City, Fukushima, Japan.
authors.forename: Hidenori
authors.initials: H
authors.lastname: Akutsu
mesh_terms: D000818:Animals; D055415:Bone Morphogenetic Protein 4 / Q000378:metabolism; D002454:Cell Differentiation / Q000502:physiology*; D002470:Cell Survival / Q000502:physiology; D002478:Cells, Cultured; D005260:Female; D020869:Gene Expression Profiling; D005819:Genetic Markers / Q000235:genetics; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000378:metabolism*; D053552:Keratin-7 / Q000378:metabolism; D051379:Mice; D008856:Microscopy, Fluorescence; D014327:Trophoblasts / Q000378:metabolism*
chemical_list: D055415:Bone Morphogenetic Protein 4; D005819:Genetic Markers; C507497:KRT7 protein, human; D053552:Keratin-7