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Effects of different donor cells on the development of nuclear-transferred porcine embryos
Ist Teil von
Yíchuán, 2007-02, Vol.29 (2), p.211
Ort / Verlag
China
Erscheinungsjahr
2007
Link zum Volltext
Quelle
MEDLINE
Beschreibungen/Notizen
This study was undertaken to systematically examine the effects of different donor cells and numbers of passages on the development of nuclear-transferred porcine embryos, so as to establish a preliminary procedure for porcine cloning. Porcine oocytes obtained at slaughter were matured in vitro for 40-44 h and then enucleated in manipulation medium containing 5 micro g/mL cytochalasin B. Fibroblast cells (FC), oviduct epithelial cells (OEC), granulosa cells (GC) and cumulus cells (CC) after 3-9 passages in 10% FBS-supplemented culture medium were either treated by serum starvation (0.5% FBS for 2-9 days), 0.1 micro g/mL aphidiconlin (APD) for 1 day and 0.5% FBS for 2-9 days or left untreated in complete medium for 2-9 days. They were transferred into enucleated oocytes by microinjection or electric fusion (100 V/mm, 30 ms and 1 pulse). Reconstituted embryos were activated with a combination of calcium ionophore A23187 or electric pulse and 6-DMAP, and cultured for 6 days, to evaluate their cleavage and embryo