Federal University of Rio Grande do Sul
Renewed interest in parthenogenetic activation of mammalian oocytes has come about for scientific, medical, and economic reasons. Germinal vesicle (GV), MI oocytes and failed to fertilize (FF) oocytes are usually discarded in human IVF cycles. However, their developmental potential may be rescued by Artificial Oocyte Activation (AOA) to generate human embryonic stem cell (hESC) lines from blastocysts obtained after AOA. In cattle industry AOA techniques have been developed to create ESC lines and to improve cloning efficiency. Various methods have been investigated for parthenogenetically activating oocytes and it seems that there is not a single method that activates oocytes from every species studied. Different groups have developed protocols optimized for their experimental objectives. The goal of this collection is to (1) provide an optimized AOA methodology for human immature and FF oocytes, to create parthenogenetic HESCs, (2) share meticulous details for manual isolation of the inner cell mass derivation in different substrates (3) provide information on AOA performed using a single or a combination of activating agents at different bovine oocyte maturation stages, as an important tool to achieve high efficiency on nuclear transfer experiments for cloning strategies. (4) share details on polimerase chain reaction for TNF-α gene expression and comet techniques to assess parthenote blastocyst viability and cell death for each activating treatment.