Embryonic stem cells (ESCs) represent a promising tool for cell therapy, regenerative medicine and tissue repair. At the same time they constitute an invaluable model for basic investigations in developmental biology, nuclear reprogramming and differentiation process. ESCs are very unique due to their unlimited self-renewal ability and high plasticity that allow them to differentiate into all embryonic tissues. However, these properties have been so far only demonstrated in the mouse and, to a lesser extent, in man. Assessment of ESC capabilities in species different from the mouse is an ongoing topic of interest and is crucial in view of their potential use as experimental models in pre-clinical applications. The mouse model is not adequate when long-term effects of cell replacement need to be evaluated. The pig has been considered for a long time among the best models for pre-clinical development of therapeutic approaches and represents an innovative model due to its morphological and functional affinity with man; therefore, pig ESCs are attracting renewed interest. However, a number of open questions need to be addressed since no validated protocols for the derivation and maintenance of pig ESCs have yet been established. In the present paper data from the literature will be presented together with experimental evidence recently obtained in our laboratory. We will discuss aspects related to the timing of isolation, the initiation of primary cultures, the use of different culture conditions and cytokines. The identification of pluripotency-related molecular markers in the pig will also be examined. Finally, the ability to respond to specifically formulated medium with spontaneous as well as induced differentiation will be assessed.
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