NEW APPROACHES TO THE GENERATION OF GENETICALLY VALUABLE RHESUS MONKEY

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This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Initial attempts to use pigtail macaques as surrogate recipients for rhesus macaque embryos showed that the efficiency with which pregnancies could be generated was too low to use this method routinely. To ensure that this was not due to inherent problems with performing embryo transfer in this species, we developed an in vitro fertilization system to generate pigtail embryos and then used those in embryo transfers into pigtail females. This resulted in the birth of two pigtail macaque infants. The focus of this project has now shifted towards using preimplantation genetic diagnosis (PGD) to genotype embryos prior to their transfer into recipients. During the past year we performed six oocyte collections from which 182 oocytes were obtained. A total of 120 of the resulting embryos were biopsied and in each case a single cell was subjected to PCR analysis to determine its sex. A few embryos from each batch were kept in culture to ensure their development had not been compromised. We currently are performing embryo transfers using sexed embryos. Initial attempts to transfer embryos back into the same female from which the oocytes had been obtained proved unsuccessful. The reasons for this are not clear but may possibly stem from a depletion of follicular cells during the aspiration process which are necessary to maintain pregnancy.