Reprod. Nutr. Dev. 44 (2004) 219-231
DOI: 10.1051/rnd:2004030

Optimisation of in vitro culture conditions in B6CBF1 mouse embryos

Bi-ke Zhu^a, Simon K. Walker^b and Simon Maddocks<sup>c

a</sup>  Faculty of Veterinary Science, University of Sydney, Camden, NSW 2570, Australia
^b  South Australian Research & Development Institute, Turretfield Research Centre, Rosedale, SA 5350, Australia
^c  South Australian Research & Development Institute, Roseworthy Campus, Roseworthy, SA 5371, Australia

(Received 10 September 2003; accepted 23 February 2004)

Abstract - This study was undertaken to investigate the effects of three media, volume and type of oil and frequency of observation on the in vitro development of mouse zygotes. B6CBF1 female mice (4 to 6 wk old) were superovulated using PMSG/hCG and mated with a proven fertile male of the same strain. Putative zygotes with polar bodies were collected from the oviducts of mated mice, 25-28 h after hCG injection, and were cultured in vitro. Embryo development was evaluated at either 96 h and 120 h or every 24 h for 120 h. The results obtained showed that the CZB medium was better than the KSOM and HCO₃HTF media, and the use of 1 mL of paraffin oil was better than the use of 0.5 mL of paraffin oil. The effect of paraffin oil and mineral oil on embryo development was examined and the results indicated that the use of paraffin oil was better than the use of mineral oil. Repeated observations did not influence the proportion of embryos developing to blastocysts.

Key words: in vitro development / media / type of oil / frequency of observation / mouse embryos

Corresponding author: Bi-ke Zhu bzhu7534@usyd.edu.au

© INRA, EDP Sciences 2004