title: Establishment of Homozygote Mutant Human Embryonic Stem Cells by Parthenogenesis.
abstract: We report on the derivation of a diploid 46(XX) human embryonic stem cell (HESC) line that is homozygous for the common deletion associated with Spinal muscular atrophy type 1 (SMA) from a pathenogenetic embryo. By characterizing the methylation status of three different imprinted loci (MEST, SNRPN and H19), monitoring the expression of two parentally imprinted genes (SNRPN and H19) and carrying out genome-wide SNP analysis, we provide evidence that this cell line was established from the activation of a mutant oocyte by diploidization of the entire genome. Therefore, our SMA parthenogenetic HESC (pHESC) line provides a proof-of-principle for the establishment of diseased HESC lines without the need for gene manipulation. As mutant oocytes are easily obtained and readily available during preimplantation genetic diagnosis (PGD) cycles, this approach should provide a powerful tool for disease modelling and is especially advantageous since it can be used to induce large or complex mutations in HESCs, including gross DNA alterations and chromosomal rearrangements, which are otherwise hard to achieve.
authors.affiliation: Stem Cell Research Laboratory, Shaare Zedek Medical Center affiliated with the Hebrew University School of Medicine, Jerusalem, Israel.
authors.forename: Silvina
authors.initials: S
authors.lastname: Epsztejn-Litman
authors.affiliation: Stem Cell Research Laboratory, Shaare Zedek Medical Center affiliated with the Hebrew University School of Medicine, Jerusalem, Israel.
authors.forename: Yaara
authors.initials: Y
authors.lastname: Cohen-Hadad
authors.affiliation: Stem Cell Research Laboratory, Shaare Zedek Medical Center affiliated with the Hebrew University School of Medicine, Jerusalem, Israel.
authors.forename: Shira
authors.initials: S
authors.lastname: Aharoni
authors.affiliation: Zohar PGD Lab, Medical Genetics Institute, Shaare Zedek Medical Center affiliated with the Hebrew University School of Medicine, Jerusalem, Israel.
authors.forename: Gheona
authors.initials: G
authors.lastname: Altarescu
authors.affiliation: Zohar PGD Lab, Medical Genetics Institute, Shaare Zedek Medical Center affiliated with the Hebrew University School of Medicine, Jerusalem, Israel.
authors.forename: Paul
authors.initials: P
authors.lastname: Renbaum
authors.affiliation: Zohar PGD Lab, Medical Genetics Institute, Shaare Zedek Medical Center affiliated with the Hebrew University School of Medicine, Jerusalem, Israel.
authors.forename: Ephrat
authors.initials: E
authors.lastname: Levy-Lahad
authors.affiliation: IVF Unit, Shaare Zedek Medical Center affiliated with the Hebrew University School of Medicine, Jerusalem, Israel.
authors.forename: Oshrat
authors.initials: O
authors.lastname: Schonberger
authors.affiliation: IVF Unit, Shaare Zedek Medical Center affiliated with the Hebrew University School of Medicine, Jerusalem, Israel.
authors.forename: Talia
authors.initials: T
authors.lastname: Eldar-Geva
authors.affiliation: Zohar PGD Lab, Medical Genetics Institute, Shaare Zedek Medical Center affiliated with the Hebrew University School of Medicine, Jerusalem, Israel.
authors.forename: Sharon
authors.initials: S
authors.lastname: Zeligson
authors.affiliation: Stem Cell Research Laboratory, Shaare Zedek Medical Center affiliated with the Hebrew University School of Medicine, Jerusalem, Israel.
authors.forename: Rachel
authors.initials: R
authors.lastname: Eiges
mesh_terms: D018392:Genomic Imprinting; D006720:Homozygote; D000066449:Human Embryonic Stem Cells / Q000378:metabolism / Q000473:pathology; D006801:Humans; D009154:Mutation; D010312:Parthenogenesis; D020641:Polymorphism, Single Nucleotide; D011506:Proteins / Q000235:genetics / Q000378:metabolism; D062085:RNA, Long Noncoding / Q000235:genetics / Q000378:metabolism; D014897:Spinal Muscular Atrophies of Childhood / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D055517:snRNP Core Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: C094335:H19 long non-coding RNA; D011506:Proteins; D062085:RNA, Long Noncoding; C526417:SNRPN protein, human; C103637:mesoderm specific transcript protein; D055517:snRNP Core Proteins