Self-organisation of the human embryo in the absence of maternal tissues

Remodelling of the human embryo at implantation is indispensable for successful pregnancy. Yet it has remained mysterious because of the experimental hurdles that beset the study of this developmental phase. Here, we establish an in vitro system to culture human embryos through implantation stages in the absence of maternal tissues and reveal the key events of early human morphogenesis. These include segregation of the pluripotent embryonic and extra-embryonic lineages, and morphogenetic rearrangements leading to generation of a bilaminar disc, formation of a pro-amniotic cavity within the embryonic lineage, appearance of the prospective yolk sac, and trophoblast differentiation. Using human embryos and human pluripotent stem cells, we show that the reorganization of the embryonic lineage is mediated by cellular polarization leading to cavity formation. Together, our results indicate that the critical remodelling events at this stage of human development are embryo-autonomous, highlighting the remarkable and unanticipated self-organizing properties of human embryos.

[1]  R. Lovell-Badge,et al.  What if stem cells turn into embryos in a dish? , 2015, Nature Methods.

[2]  R. Edwards,et al.  Early Stages of Fertilization in vitro of Human Oocytes Matured in vitro , 1969, Nature.

[3]  W. Luckett,et al.  The development of primordial and definitive amniotic cavities in early Rhesus monkey and human embryos. , 1975, The American journal of anatomy.

[4]  P. Bourgine,et al.  A workflow to process 3D+time microscopy images of developing organisms and reconstruct their cell lineage , 2016, Nature Communications.

[5]  K. Mostov,et al.  From cells to organs: building polarized tissue , 2008, Nature Reviews Molecular Cell Biology.

[6]  J. Brosens,et al.  Molecular aspects of implantation failure. , 2012, Biochimica et biophysica acta.

[7]  Roger A. Pedersen,et al.  Human preimplantation embryo development , 2022 .

[8]  Kevin Eggan,et al.  Analysis of human embryos from zygote to blastocyst reveals distinct gene expression patterns relative to the mouse. , 2013, Developmental biology.

[9]  J. Rock,et al.  A description of 34 human ova within the first 17 days of development. , 1956, The American journal of anatomy.

[10]  R. Roberts,et al.  Low O2 tensions and the prevention of differentiation of hES cells. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[11]  N. Macklon,et al.  In-vitro model systems for the study of human embryo-endometrium interactions. , 2013, Reproductive biomedicine online.

[12]  M. Zernicka-Goetz,et al.  Culture of human embryos through implantation stages in vitro , 2016 .

[13]  C. Jones,et al.  Studies of mesenchymal cells from 1st trimester human placenta: expression of cytokeratin outside the trophoblast lineage. , 1999, Placenta.

[14]  M. Zernicka-Goetz,et al.  Developmental plasticity, cell fate specification and morphogenesis in the early mouse embryo , 2014, Philosophical Transactions of the Royal Society B: Biological Sciences.

[15]  Madeline A. Lancaster,et al.  Cerebral organoids model human brain development and microcephaly , 2013, Nature.

[16]  James Peter Hill.  Croonian Lecture: The Developmental History of the Primates , 1932 .

[17]  DEVELOPMENTAL HISTORY OF THE PRIMATES , 1930, British medical journal.

[18]  J. Palis,et al.  Yolk-sac hematopoiesis: the first blood cells of mouse and man. , 2001, Experimental hematology.

[19]  Apoorva Mandavilli,et al.  Of mice and men , 2006, Nature Medicine.

[20]  Magdalena Zernicka-Goetz,et al.  Self-Organizing Properties of Mouse Pluripotent Cells Initiate Morphogenesis upon Implantation , 2014, Cell.

[21]  J. Deprest,et al.  On the origin of amniotic stem cells: of mice and men. , 2010, The International journal of developmental biology.

[22]  A. Hendrickx,et al.  Differentiation of the embryonic disc, amnion, and yolk sac in the rhesus monkey. , 1986, The American journal of anatomy.

[23]  Jin-ho Lim,et al.  Effect of artificial shrinkage on clinical outcome in fresh blastocyst transfer cycles , 2011, Clinical and experimental reproductive medicine.

[24]  Y. Shao,et al.  Lumen Formation Is an Intrinsic Property of Isolated Human Pluripotent Stem Cells , 2015, Stem cell reports.

[25]  Vanessa J. Jones,et al.  Culture of whole mouse embryos at early postimplantation to organogenesis stages: developmental staging and methods. , 2010, Methods in enzymology.

[26]  Genee Y. Lee,et al.  Three-dimensional culture models of normal and malignant breast epithelial cells , 2007, Nature Methods.

[27]  Brian Keith,et al.  HIF-2alpha regulates Oct-4: effects of hypoxia on stem cell function, embryonic development, and tumor growth. , 2006, Genes & development.

[28]  D. Gardner.  The impact of physiological oxygen during culture, and vitrification for cryopreservation, on the outcome of extended culture in human IVF. , 2016, Reproductive biomedicine online.

[29]  A. Trounson,et al.  Human embryonic stem cells: prospects for development , 2004, Development.

[30]  R. Weinstein,et al.  Prospects for Development , 1999 .

[31]  M. Zernicka-Goetz,et al.  In vitro culture of mouse blastocysts beyond the implantation stages , 2014, Nature Protocols.

[32]  J. Goeman,et al.  Tracking the progression of the human inner cell mass during embryonic stem cell derivation , 2012, Nature Biotechnology.

[33]  K. Niakan,et al.  Human pre-implantation embryo development , 2012, Development.

[34]  Samantha A. Morris,et al.  Dynamics of anterior–posterior axis formation in the developing mouse embryo , 2012, Nature Communications.

[35]  S. M. Chuva de Sousa Lopes,et al.  Cell Lineage Specific Distribution of H3K27 Trimethylation Accumulation in an In Vitro Model for Human Implantation , 2012, PloS one.

[36]  Z. Werb,et al.  A molecular switch for the orientation of epithelial cell polarization. , 2014, Developmental cell.

[37]  R. Edwards,et al.  Fertilization and Cleavage in vitro of Preovulator Human Oocytes , 1970, Nature.

[38]  J. Nichols,et al.  Human hypoblast formation is not dependent on FGF signalling , 2012, Developmental biology.