caa a22 4500 445817763 CHVBK 20180317145234.0 cr unu---uuuuu 170323e20110501xx s 000 0 eng 10.1007/s10815-011-9540-x doi (NATIONALLICENCE)springer-10.1007/s10815-011-9540-x Use of parthenogenetic activation of human oocytes as an experimental model for evaluation of polar body based PGD assay performance [Elektronische Daten] [Alessio Paffoni, Valentina Paracchini, Stefania Ferrari, Claudia Scarduelli, Manuela Seia, Domenico Coviello, Guido Ragni] Purpose: 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, andvalidated 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. Springer Science+Business Media, LLC, 2011 CFTR nationallicence HBB nationallicence Monogenic disease nationallicence Genetic diagnosis nationallicence Paffoni Alessio Department of Obstetrics and Gynaecology, Infertility Unit, Fondazione IRCCS Ca' Granda Ospedale Maggiore Policlinico, via M. Fanti 6, 20122, Milan, Italy aut Paracchini Valentina Fondazione IRCCS Ca' Granda Ospedale Maggiore Policlinico, Medical Genetics Laboratory, Milan, Italy aut Ferrari Stefania Department of Obstetrics and Gynaecology, Infertility Unit, Fondazione IRCCS Ca' Granda Ospedale Maggiore Policlinico, via M. Fanti 6, 20122, Milan, Italy aut Scarduelli Claudia Department of Obstetrics and Gynaecology, Infertility Unit, Fondazione IRCCS Ca' Granda Ospedale Maggiore Policlinico, via M. Fanti 6, 20122, Milan, Italy aut Seia Manuela Fondazione IRCCS Ca' Granda Ospedale Maggiore Policlinico, Medical Genetics Laboratory, Milan, Italy aut Coviello Domenico Fondazione IRCCS Ca' Granda Ospedale Maggiore Policlinico, Medical Genetics Laboratory, Milan, Italy aut Ragni Guido Department of Obstetrics and Gynaecology, Infertility Unit, Fondazione IRCCS Ca' Granda Ospedale Maggiore Policlinico, via M. Fanti 6, 20122, Milan, Italy aut Journal of Assisted Reproduction and Genetics Springer US; http://www.springer-ny.com 28/5(2011-05-01), 461-470 1058-0468 28:5<461 2011 28 10815 https://doi.org/10.1007/s10815-011-9540-x text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s10815-011-9540-x text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Paffoni Alessio Department of Obstetrics and Gynaecology, Infertility Unit, Fondazione IRCCS Ca' Granda Ospedale Maggiore Policlinico, via M. Fanti 6, 20122, Milan, Italy aut NATIONALLICENCE 700 1- Paracchini Valentina Fondazione IRCCS Ca' Granda Ospedale Maggiore Policlinico, Medical Genetics Laboratory, Milan, Italy aut NATIONALLICENCE 700 1- Ferrari Stefania Department of Obstetrics and Gynaecology, Infertility Unit, Fondazione IRCCS Ca' Granda Ospedale Maggiore Policlinico, via M. Fanti 6, 20122, Milan, Italy aut NATIONALLICENCE 700 1- Scarduelli Claudia Department of Obstetrics and Gynaecology, Infertility Unit, Fondazione IRCCS Ca' Granda Ospedale Maggiore Policlinico, via M. Fanti 6, 20122, Milan, Italy aut NATIONALLICENCE 700 1- Seia Manuela Fondazione IRCCS Ca' Granda Ospedale Maggiore Policlinico, Medical Genetics Laboratory, Milan, Italy aut NATIONALLICENCE 700 1- Coviello Domenico Fondazione IRCCS Ca' Granda Ospedale Maggiore Policlinico, Medical Genetics Laboratory, Milan, Italy aut NATIONALLICENCE 700 1- Ragni Guido Department of Obstetrics and Gynaecology, Infertility Unit, Fondazione IRCCS Ca' Granda Ospedale Maggiore Policlinico, via M. Fanti 6, 20122, Milan, Italy aut NATIONALLICENCE 773 0- Journal of Assisted Reproduction and Genetics Springer US; http://www.springer-ny.com 28/5(2011-05-01), 461-470 1058-0468 28:5<461 2011 28 10815 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer