Morphology and biochemistry of in-vitro produced bovine embryos: implications for their cryopreservation

Massip, A.; Mermillod, Pascal; Dinnyés, András

doi:10.1093/oxfordjournals.humrep.a135837

Cited by 149 publications

(82 citation statements)

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“…However, in vitro produced (IVP) embryos are more sensitive to chilling (31) and freezing than their in vivo countea'parts (16,20). The differences between in vitro and in vivo produced embryos have been investigated by several authors (13,36).…”

Section: ~Troductionmentioning

confidence: 99%

Delipidating in vitro-produced bovine zygotes: Effect on further development and consequences for freezability

Díez

Heyman

Bourhis³

et al. 2001

Theriogenology

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To study the effect of partial removal of intracytoplasmatic lipids from bovine zygotes on their in vitro and in vivo survival, presumptive zygotes were delipidated by micromanipulation and cocultured with Veto cells in B2+10% FCS. Blastocyst rates of delipidated (n=960), sham (centrifuged but not delipidated, n=830) and control embryos (n=950) were 42.1, 42.3 and 39.9% respectively (P > 0.05). Day 7 blastocysts derived from delipidated zygotes had a mean of 123.9 + 45.6 nuclei compared to 137.5 + 32.9 for control blastocysts (P > 0.05). The full-term development of delipidated blastocysts after single transfer to recipients was similar to that of control IVF blastocysts (41.2% vs 45.4% respectively). To assess the effect of delipidation on the embryo tolerance to freczing/thawing, delipidated (n=73), control (n=67) and shmn (n=50) Day 7 blastocysts were frozen in 1.36 M glycerol + 0.25 M sucrose in PBS. After thawing, embryos were cocultured for 72 h with Vero cells in B2+10% FCS. Survival rates at 24 h were not significantly different between groups. However, in the delipidated group, the survival rate after 48 h in culture was significantly higher than in the control group (56.2 vs 39.8, P < 0.02), resulting in a higher hatching rate after 3 days in culture (45.2 vs 22.4, P < 0.02). Pregnancy rates for delipidated and control frozen/thawed embryos were respectively 10.5 and 22.2% (P > 0.05). Electron microscopic observations showed much fewer lipid droplets (and smaller) in delipated blastocysts than in controls. Taken together, our data show that delipidation of one cell stage bovine embryos is compatible with their normal development to term and has a beneficial effect on their tolerance to freezing and thawing at the blastocyst stage. This procedure, however, alters the developmental potential of such blastocysts, suggesting that maternally inherited lipid stores interfere with metabolic recovery after thawing.

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Section: ~Troductionmentioning

confidence: 99%

Delipidating in vitro-produced bovine zygotes: Effect on further development and consequences for freezability

Díez

Heyman

Bourhis³

et al. 2001

Theriogenology

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“…Improvement of embryo survival after freezing can be achieved by changing the conditions of in vitro culture, and/or modifying ''standard'' cryopreservation procedures [14].…”

Section: Introductionmentioning

confidence: 99%

“…Slow freezing of in vitro produced embryos reduced post thaw survival rates compared with their in vivo counterparts, mostly due to their susceptibility to ice crystal formation. Cryopreservation of embryos by vitrification yields higher survival than conventional slow freezing [14,[29][30][31].…”

Section: Introductionmentioning

confidence: 99%

Serum free embryo culture medium improves in vitro survival of bovine blastocysts to vitrification

et al. 2008

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The aim of this study was to examine the effects of co-culture with Vero cells during the in vitro maturation (IVM) and three culture media, B2 + 5% fetal calf serum (FCS) on Vero cells, synthetic oviduct fluid (SOF) + 5% FCS, and SOF + 20 g L À1 bovine serum albumin (BSA), on the developmental competence of the embryos and their ability to survive vitrification/warming. We also tested the effect of morphological quality and the age of the embryo on its sensitivity to vitrification. The IVM system neither affects the embryo development up to Day 7 nor survival rates after vitrification. The culture of embryos in SOF + FCS and in Vero cells + B2 allowed obtaining more Day 6 and Day 7 blastocysts, and a higher % of Day 7 blastocysts vitrified than culture in SOF + BSA. Contrarily, on Day 8, more blastocysts were vitrified in SOF + BSA than in SOF + FCS. Blastocysts quality affected development after vitrification/warming, and Day 7 embryos showed higher survival rates than their Day 8 counterparts. Day 7 blastocysts produced in Vero cells or in SOF + BSA survived at higher rates than those produced in SOF + FCS at 24 and 48 h after warming. Embryo culture with BSA allows obtaining hatching rates after vitrification/warming higher than those obtained after coculture with Vero cells in B2 and FCS. Moreover, this system provides hatching rates from Day 8 blastocysts comparable to those obtained on Day 7 in Vero cells. Further studies, including embryo transfer to recipients, are needed to clarify factors affecting the freezability of in vitro produced bovine embryos. #

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“…This could be related to the embryo density in the culture medium since good results were reported culturing one embryo per pL of medium under mineral oil (Ferry et al, 1994 (Ferry et al, 1994;Massip et al, 1995).…”

mentioning

confidence: 99%

“…Recently, it was demonstrated that the production of bovine blastocysts was also possible in a simple defined medium like SOF, although the concentration of O Z (5%) is critical since a reduced embryo development at high oxygen concentration is observed (Carolan et al, 1995;Massip et al, 1995). The low oxygen tension is thought to act by reducing auto-oxidative damage to which the embryos are susceptible.…”

mentioning

confidence: 99%

Beneficial effects of Vero cells for developing IVF bovine eggs in two different coculture systems

Menck¹,

Guyader-Joly²,

Peynot³

et al. 1997

Reprod. Nutr. Dev.

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Morphology and biochemistry of in-vitro produced bovine embryos: implications for their cryopreservation

Cited by 149 publications

References 0 publications

Delipidating in vitro-produced bovine zygotes: Effect on further development and consequences for freezability

Delipidating in vitro-produced bovine zygotes: Effect on further development and consequences for freezability

Serum free embryo culture medium improves in vitro survival of bovine blastocysts to vitrification

Beneficial effects of Vero cells for developing IVF bovine eggs in two different coculture systems

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