Derivation of new pluripotent stem cells from human extended pluripotent stem cells with formative features and trophectoderm potential.

Previous studies have demonstrated the existence of intermediate stem cells, which have been successfully obtained from human naive pluripotent stem cells (PSCs) and peri-implantation embryos. However, it is not known whether human extended pluripotent stem cells (hEPSCs) can be directly induced into intermediate stem cells. Moreover, the ability of extra-embryonic lineage differentiation in intermediate stem cells has not been verified. In this issue, we transformed hEPSCs into a kind of novel intermediate pluripotent stem cell resembling embryonic days 8-9 (E8-E9) epiblasts and proved its feature of formative epiblasts. We engineered hEPSCs from primed hPSCs under N2B27-LCDM (N2B27 plus Lif, CHIR, DiH and MiH) conditions. Then, we added Activin A, FGF and XAV939 to modulate signalling pathways related to early humans' embryogenesis. We performed RNA-seq and CUT&Tag analysis to compare with AF9-hPSCs from different pluripotency stages of hPSCs. Trophectoderm (TE), primordial germ cells-like cells (PGCLC) and endoderm, mesoderm, and neural ectoderm induction were conducted by specific small molecules and proteins. AF9-hPSCs transcription resembled that of E8-E9 peri-implantation epiblasts. Signalling pathway responsiveness and histone methylation further revealed their formative pluripotency. Additionally, AF9-hPSCs responded directly to primordial germ cells (PGCs) specification and three germ layer differentiation signals in vitro. Moreover, AF9-hPSCs could differentiate into the TE lineage. Therefore, AF9-hPSCs represented an E8-E9 formative pluripotency state between naïve and primed pluripotency, opening new avenues for studying human pluripotency development during embryogenesis.

[1]  S. Liang,et al.  Schisanhenol improves early porcine embryo development by regulating the phosphorylation level of MAPK. , 2021, Theriogenology.

[2]  Xiaochen Bo,et al.  clusterProfiler 4.0: A universal enrichment tool for interpreting omics data , 2021, Innovation.

[3]  Alexander W. Bruce,et al.  DDX21 is a p38-MAPK-sensitive nucleolar protein necessary for mouse preimplantation embryo development and cell-fate specification , 2021, Open Biology.

[4]  K. Woltjen,et al.  Capturing human trophoblast development with naive pluripotent stem cells in vitro. , 2021, Cell stem cell.

[5]  J. Nichols,et al.  Human naive epiblast cells possess unrestricted lineage potential , 2021, Cell stem cell.

[6]  Wei Xie,et al.  Formative pluripotent stem cells show features of epiblast cells poised for gastrulation , 2021, Cell Research.

[7]  Mei-lin Xu,et al.  High‐throughput screening in postimplantation haploid epiblast stem cells reveals Hs3st3b1 as a modulator for reprogramming , 2021, Stem cells translational medicine.

[8]  Yi Zhang,et al.  M2 macrophage‐derived G‐CSF promotes trophoblasts EMT, invasion and migration via activating PI3K/Akt/Erk1/2 pathway to mediate normal pregnancy , 2021, Journal of cellular and molecular medicine.

[9]  Stéphanie Kilens,et al.  Induction of Human Trophoblast Stem Cells from Somatic Cells and Pluripotent Stem Cells. , 2020, Cell reports.

[10]  Haixi Sun,et al.  Derivation of Intermediate Pluripotent Stem Cells Amenable to Primordial Germ Cell Specification. , 2020, Cell stem cell.

[11]  J. Nichols,et al.  Capture of Mouse and Human Stem Cells with Features of Formative Pluripotency , 2020, bioRxiv.

[12]  René A. M. Dirks,et al.  In vitro capture and characterization of embryonic rosette-stage pluripotency between naive and primed states , 2020, Nature Cell Biology.

[13]  Hao Zhang,et al.  In vitro testicular organogenesis from human fetal gonads produces fertilization-competent spermatids , 2020, Cell Research.

[14]  Ting Wang,et al.  Derivation of trophoblast stem cells from naïve human pluripotent stem cells , 2020, eLife.

[15]  Yun Zheng,et al.  A developmental landscape of 3D-cultured human pre-gastrulation embryos , 2019, Nature.

[16]  Steven L Salzberg,et al.  Graph-based genome alignment and genotyping with HISAT2 and HISAT-genotype , 2019, Nature Biotechnology.

[17]  F. Tang,et al.  Reconstituting the transcriptome and DNA methylome landscapes of human implantation , 2019, Nature.

[18]  Lia S. Campos,et al.  Establishment of porcine and human expanded potential stem cells , 2019, Nature Cell Biology.

[19]  Satu Kuure,et al.  MAPK/ERK Signaling in Regulation of Renal Differentiation , 2019, International journal of molecular sciences.

[20]  Jia Gu,et al.  fastp: an ultra-fast all-in-one FASTQ preprocessor , 2018, bioRxiv.

[21]  M. Suyama,et al.  Derivation of Human Trophoblast Stem Cells. , 2018, Cell stem cell.

[22]  Aleksandra A. Kolodziejczyk,et al.  Establishment of mouse expanded potential stem cells , 2017, Nature.

[23]  Paul Bertone,et al.  Epigenetic resetting of human pluripotency , 2017, Development.

[24]  D. Charnock-Jones,et al.  RNA-seq reveals conservation of function among the yolk sacs of human, mouse, and chicken , 2017, Proceedings of the National Academy of Sciences.

[25]  Xiang Li,et al.  Derivation of Pluripotent Stem Cells with In Vivo Embryonic and Extraembryonic Potency , 2017, Cell.

[26]  Olivier Pourquié,et al.  Generation of human muscle fibers and satellite-like cells from human pluripotent stem cells in vitro , 2016, Nature Protocols.

[27]  Pang Wei Koh,et al.  Mapping the Pairwise Choices Leading from Pluripotency to Human Bone, Heart, and Other Mesoderm Cell Types , 2016, Cell.

[28]  J. Nichols,et al.  Naive Pluripotent Stem Cells Derived Directly from Isolated Cells of the Human Inner Cell Mass , 2016, Stem cell reports.

[29]  Qinghua Shi,et al.  Complete Meiosis from Embryonic Stem Cell-Derived Germ Cells In Vitro. , 2016, Cell stem cell.

[30]  G. Daley,et al.  Hallmarks of pluripotency , 2015, Nature.

[31]  S. Yamanaka,et al.  Robust In Vitro Induction of Human Germ Cell Fate from Pluripotent Stem Cells. , 2015, Cell stem cell.

[32]  M. Azim Surani,et al.  SOX17 Is a Critical Specifier of Human Primordial Germ Cell Fate , 2015, Cell.

[33]  W. Huber,et al.  Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2 , 2014, Genome Biology.

[34]  R. Young,et al.  Systematic Identification of Culture Conditions for Induction and Maintenance of Naive Human Pluripotency , 2014, Cell stem cell.

[35]  J. Nichols,et al.  The ability of inner-cell-mass cells to self-renew as embryonic stem cells is acquired following epiblast specification , 2014, Nature Cell Biology.

[36]  Fidel Ramírez,et al.  deepTools: a flexible platform for exploring deep-sequencing data , 2014, Nucleic Acids Res..

[37]  Angelique M. Nelson,et al.  Derivation of naïve human embryonic stem cells , 2014, Proceedings of the National Academy of Sciences.

[38]  I. Weissman,et al.  Efficient endoderm induction from human pluripotent stem cells by logically directing signals controlling lineage bifurcations. , 2014, Cell stem cell.

[39]  I. Amit,et al.  Derivation of novel human ground state naive pluripotent stem cells , 2013, Nature.

[40]  Hélène Touzet,et al.  SortMeRNA: fast and accurate filtering of ribosomal RNAs in metatranscriptomic data , 2012, Bioinform..

[41]  Russell J. Taylor,et al.  Wnt/β-catenin signaling promotes differentiation, not self-renewal, of human embryonic stem cells and is repressed by Oct4 , 2012, Proceedings of the National Academy of Sciences.

[42]  Steven L Salzberg,et al.  Fast gapped-read alignment with Bowtie 2 , 2012, Nature Methods.

[43]  Mark D. Robinson,et al.  edgeR: a Bioconductor package for differential expression analysis of digital gene expression data , 2009, Bioinform..

[44]  J. Nichols,et al.  Naive and primed pluripotent states. , 2009, Cell stem cell.

[45]  M. Tomishima,et al.  Highly efficient neural conversion of human ES and iPS cells by dual inhibition of SMAD signaling , 2009, Nature Biotechnology.

[46]  M. Trotter,et al.  Derivation of pluripotent epiblast stem cells from mammalian embryos , 2007, Nature.

[47]  R. McKay,et al.  New cell lines from mouse epiblast share defining features with human embryonic stem cells , 2007, Nature.