title: Use of parthenogenetic activation of human oocytes as an experimental model for evaluation of polar body based PGD assay performance.
abstract: OBJECTIVE
  To develop an experimental model to assess the feasibility of polar body preimplantation genetic diagnosis without requiring oocyte fertilization.
  
  
  METHODS
  First polar body was removed from donated oocytes and second polar body was biopsied after parthenogenetic activation. Molecular analysis on both polar bodies involved a fluorescent multiplex polymerase chain reaction of short-tandem repeat markers, closely linked to genes of interest. Main outcome measures were: allele segregation through polar bodies and haploid nucleus, recombination rates between alleles and frequency of Allele Drop Out.
  
  
  RESULTS
  Twenty-six out of 39 oocytes extruded a second polar body after activation. Ninety-two percent of the first polar bodies and 20 out of 26 (77%) second polar bodies were successfully amplified. Eighty percent of first polar bodies were heterozygous for CFTR and 55% for HBB. Analysis of second polar bodies predicted the genotype of the oocytes in case of heterozygous first polar body, and validated results in homozygous cases. Frequency of allele drop out was 4%.
  
  
  CONCLUSIONS
  Our model confirms that polar body preimplantation genetic diagnosis for single gene disorders can be evaluated using parthenogenetic oocytes and offers an option to set up procedures without requiring oocyte fertilization.
authors.affiliation: Department of Obstetrics and Gynaecology, Infertility Unit, Fondazione IRCCS Ca' Granda Ospedale Maggiore Policlinico, via M. Fanti 6, 20122 Milan, Italy. alessio.paffoni@alice.it
authors.forename: Alessio
authors.initials: A
authors.lastname: Paffoni
authors.forename: Valentina
authors.initials: V
authors.lastname: Paracchini
authors.forename: Stefania
authors.initials: S
authors.lastname: Ferrari
authors.forename: Claudia
authors.initials: C
authors.lastname: Scarduelli
authors.forename: Manuela
authors.initials: M
authors.lastname: Seia
authors.forename: Domenico A
authors.initials: DA
authors.lastname: Coviello
authors.forename: Guido
authors.initials: G
authors.lastname: Ragni
mesh_terms: D000483:Alleles; D005240:Feasibility Studies; D005260:Female; D005838:Genotype; D006801:Humans; D060885:Multiplex Polymerase Chain Reaction; D009865:Oocytes / Q000502:physiology*; D010312:Parthenogenesis; D059705:Polar Bodies / Q000502:physiology*; D019836:Preimplantation Diagnosis / Q000379:methods