Obtaiment of pudu (Pudu pudu) deer embryos by the somatic nuclear transfer technique

SUMMARY: The objective of this project is to produce pudu embryos, obtained by means of fibroblast transfer taken from the ear of one animal, to the oocytes of a domestic ruminant such as cattle. In further work, the pudu embryos will be transferred to the uterus of synchronized recipient females of another species. Biopsies 1mm in size were obtained from the outer rim of the ear of two male pudu deer belonging to the Buin Zoological park, Santiago, Chile. The cell lines have been established and kept according to protocols used in cattle. The oocytes are obtained through a puncture in the oocyte-cumulus complex (OCC) from cow ovaries recovered from the butcher. Each oocyte is enucleated and fused with an isolated fibroblast which is inserted beneath the pellucid zone. The fusion of cell membranes is achieved by means of electric shocks. In respect to the timetable, we have observed that a stage of two blastomers is reached on the second day, and morulae of 8 to 16 cells on the third