Derivation and characterization of pluripotent porcine cells

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Despite considerable world wide research efforts stable pluripotent embryonicstem cell (ESC) lines could not yet be derived in the domestic species. While someprogress has been achieved with the derivation of porcine embryonic germ cells (pEGC),derived cells were not able to form germ line chimeras. Here, we will use a novelapproach to produce germ line competent porcine ESC by preventing the differentiationof inner cell mass (ICM) and/or epiblast cells in culture by doxycycltne regulatedexpression of exogenous pluripotency genes. Additionally, we will use the recentprogress in the generation of induced pluripotent stem cells (iPSC) to improvereprogramming of pEGCs to complete pluripotency via induced expression of thetranscription factors 0CT4, S0X2, and NANOG, and investigate the effect of epigeneticreprogramming on the efficiency of chimera production. Putative pluripotent cells fromboth sources will be characterized by morphology, in vitro culture growth, expressionprofile of pluripotency genes, differentiation potential, and tumorigenic potential. Weanticipate that this study will result in the establishment of porcine pluripotent cell linesthat can be used for the production of germ line chimeras, and will thus constitute anovel tool for improving the efficiency of transgenic pig production with great impact onhuman and animal health.