Comparison of the effect of two different handling media on rabbit zygote developmental ability

Abstract

-Despite the attention paid to culture media, the relevance of the handling medium at egg recovery/transfer is frequently overlooked. In the present work, we compare the effect of two different handling media (PBS and HEPES-buffered Ham F10, both supplemented with 20% (v/v) FCS), upon in vitro and in vivo developmental ability of in vivo fertilised rabbit zygotes. Zygotes recovered in HEPES-buffered medium (permanence 1 h as maximum) and subsequently cultured in vitro developed more efficiently to the compacted morula (100%) and blastocyst stage (92%) than those recovered in PBS (83% and 76%, respectively, P < 0.05). Zygotes recovered in such media were then further bilaterally transferred to recipient does following a brief in vitro culture period (for 4 hours). At caesarean section (day 28 of pregnancy), significant differences were observed in both the percentage of pregnant uterine horns (PBS: 60% vs. HEPES-buffered Ham F10: 100%) and live birth rates (PBS: 14% vs. HEPES-buffered Ham F10: 34%). Thus when early rabbit zygotes must be handled, even for short incubation periods, the medium is not innocuous. culture media / buffer system / development / zygote / rabbit Résumé -Comparaison entre deux milieux de récupération et de manipulation des ovocytes de lapine sur leur capacité ultérieure de développement. Malgré l'attention prêtée aux milieux de culture, il est fréquent d'oublier l'importance du milieu de manipulation lors de la récupération ou du transfert des oeufs. Dans ce travail, nous comparons l'effet exercé par deux milieux de manipulation différents (PBS et HEPES-buffered Ham's F10, tous deux additionnés de 20 % SVF) sur la capacité de développement in vivo et in vitro des zygotes fécondés in vivo. Les zygotes récupérés dans le milieu tamponné par l'HEPES (1 h de permanence au maximum) et cultivés in vitro présentaient un meilleur développement jusqu'aux stades morula (100 %) et blastocyste (92 %) que les zygotes récupérés dans le PBS (83 % et 76 %, respectivement, P < 0,05). Dans la seconde expérience, l'effet du milieu Reprod. Nutr. Dev. 41 (2001) [181][182][183][184][185][186] 181

Effect of different culture systems on mRNA expression in developing rabbit embryos

“…Finally, mRNA expression of all in vitro developed groups was compared with two control groups consisting of 4-and 6-day in vivo developed embryos. 1993; Escribá et al, 2001;Sultana et al, 2009). However, these systems do not mimic the uterine environment and in vitro developed embryos differ from their in vivo counterparts.…”

Osmolarity and composition of cell culture media affect further development and survival in zebrafish embryos

“…Surprisingly, this toxic effect did not affect embryo cell survival during in vitro culture. Some negative effects of components of the culture media on embryos have been described in mammals, such as those due to phosphates (Escribá et al, 2001) or phenol red, although the latter is recommended precisely as a tracer for some substances in intraembryonic microinjection experiments in zebrafish (Gilmour et al, 2002). Although Sawant et al (2001) indicate that higher salinity impaired the nuclear division of the embryonic cells in zebrafish embryos, in our case L-15 medium, which is usually used in fish cell culture, can support embryonic development, even early larvae, although that medium finally caused motility loss and developmental inhibition.…”

Embryo Manipulation Techniques in the Rabbit

“…Normal embryos can be maintained under in vitro conditions or cryopreserved until transferred. Zygotes have been successfully developed into blastocyst in vitro [10,31,[104][105][106][107][108]. One example of culture media is TCM199 supplemented with 0.1% of BSA and culture is performed in 500 μl of medium layered under paraffin oil at 38.5°C, 5% CO 2 and saturated humidity [109].…”