title: Derivation and differentiation of haploid human embryonic stem cells.
abstract: Diploidy is a fundamental genetic feature in mammals, in which haploid cells normally arise only as post-meiotic germ cells that serve to ensure a diploid genome upon fertilization. Gamete manipulation has yielded haploid embryonic stem (ES) cells from several mammalian species, but haploid human ES cells have yet to be reported. Here we generated and analysed a collection of human parthenogenetic ES cell lines originating from haploid oocytes, leading to the successful isolation and maintenance of human ES cell lines with a normal haploid karyotype. Haploid human ES cells exhibited typical pluripotent stem cell characteristics, such as self-renewal capacity and a pluripotency-specific molecular signature. Moreover, we demonstrated the utility of these cells as a platform for loss-of-function genetic screening. Although haploid human ES cells resembled their diploid counterparts, they also displayed distinct properties including differential regulation of X chromosome inactivation and of genes involved in oxidative phosphorylation, alongside reduction in absolute gene expression levels and cell size. Surprisingly, we found that a haploid human genome is compatible not only with the undifferentiated pluripotent state, but also with differentiated somatic fates representing all three embryonic germ layers both in vitro and in vivo, despite a persistent dosage imbalance between the autosomes and X chromosome. We expect that haploid human ES cells will provide novel means for studying human functional genomics and development.
authors.affiliation: The Azrieli Center for Stem Cells and Genetic Research, Department of Genetics, Silberman Institute of Life Sciences, The Hebrew University, Jerusalem 91904, Israel.
authors.forename: Ido
authors.initials: I
authors.lastname: Sagi
authors.affiliation: Department of Pediatrics, Columbia University, New York, New York 10032, USA.
authors.forename: Gloryn
authors.initials: G
authors.lastname: Chia
authors.affiliation: The Azrieli Center for Stem Cells and Genetic Research, Department of Genetics, Silberman Institute of Life Sciences, The Hebrew University, Jerusalem 91904, Israel.
authors.forename: Tamar
authors.initials: T
authors.lastname: Golan-Lev
authors.affiliation: The Azrieli Center for Stem Cells and Genetic Research, Department of Genetics, Silberman Institute of Life Sciences, The Hebrew University, Jerusalem 91904, Israel.
authors.forename: Mordecai
authors.initials: M
authors.lastname: Peretz
authors.affiliation: The Azrieli Center for Stem Cells and Genetic Research, Department of Genetics, Silberman Institute of Life Sciences, The Hebrew University, Jerusalem 91904, Israel.
authors.forename: Uri
authors.initials: U
authors.lastname: Weissbein
authors.affiliation: Department of Pediatrics, Columbia University, New York, New York 10032, USA.
authors.forename: Lina
authors.initials: L
authors.lastname: Sui
authors.affiliation: Center for Women's Reproductive Care, College of Physicians and Surgeons, Columbia University, New York, New York 10019, USA.
authors.forename: Mark V
authors.initials: MV
authors.lastname: Sauer
authors.affiliation: The Azrieli Center for Stem Cells and Genetic Research, Department of Genetics, Silberman Institute of Life Sciences, The Hebrew University, Jerusalem 91904, Israel.
authors.forename: Ofra
authors.initials: O
authors.lastname: Yanuka
authors.affiliation: Department of Pediatrics, Columbia University, New York, New York 10032, USA.|The New York Stem Cell Foundation Research Institute, New York, New York 10032, USA.
authors.forename: Dieter
authors.initials: D
authors.lastname: Egli
authors.affiliation: The Azrieli Center for Stem Cells and Genetic Research, Department of Genetics, Silberman Institute of Life Sciences, The Hebrew University, Jerusalem 91904, Israel.
authors.forename: Nissim
authors.initials: N
authors.lastname: Benvenisty
mesh_terms: D002454:Cell Differentiation; D000066673:Cell Self Renewal; D002469:Cell Separation; D048429:Cell Size; D041321:Chromosomes, Human, X / Q000235:genetics; D004171:Diploidy; D015536:Down-Regulation / Q000235:genetics; D017353:Gene Deletion; D056726:Genetic Association Studies / Q000379:methods*; D005855:Germ Layers / Q000166:cytology; D006238:Haploidy; D000066449:Human Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism*; D006801:Humans; D007621:Karyotyping; D009865:Oocytes / Q000378:metabolism; D010085:Oxidative Phosphorylation; D010312:Parthenogenesis; D039904:Pluripotent Stem Cells / Q000166:cytology / Q000378:metabolism; D049951:X Chromosome Inactivation / Q000235:genetics