Comparison of two co-culture systems to assess the survival of in vitro produced bovine blastocysts after vitrification

Kaidi, Safia; Donnay, Isabelle; Langendonckt, Anne Van; Dessy, F.; Massip, A.

doi:10.1016/s0378-4320(98)00089-x

Cited by 41 publications

(31 citation statements)

References 27 publications

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“…Thus, embryos survived at higher rates when cultured in BRL than granulosa cells, and 20% O2 led to higher hatching rates than 5% O2 in co-culture. Finally, these authors conclude that the use of a co-culture system for warmed embryos ensures the best system to obtain better survival rates [13,49].…”

Section: Discussionmentioning

confidence: 91%

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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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Section: Discussionmentioning

confidence: 91%

“…The cell type used in co-culture and incubation conditions after vitrification and warming may influence blastocyst survival and quality after cryopreservation Kaidi et al [49]. Thus, embryos survived at higher rates when cultured in BRL than granulosa cells, and 20% O2 led to higher hatching rates than 5% O2 in co-culture.…”

Section: Discussionmentioning

confidence: 99%

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

et al. 2008

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“…However, there seen a significant drop in term of hatching and apoptosis rates of IVC + /PW − vs. IVC + − blastocysts compared to all the other culture conditions (p<0.05). Due to well-known correlation between the culture condition and the quality of the embryos following cryopreservation [6], it seems that addition of βME antioxidant during early stage of in vitro embryo development but not after freezing/warming has increased the chance of more embryos, even those of fair-quality, to develop to the blastocyst stage. However, following stressful process of freezing, some of these fair-quality embryos did not survive and fail to hatch and hence degenerate.…”

Section: Discussionmentioning

confidence: 99%

“…However, the survival of cryopreserved in vitro produced (IVP) embryos, as measured either by post-warming survival in culture or by established pregnancies after embryo transfer, has lagged behind that of in vivo-derived embryos [5][6][7][8].…”

mentioning

confidence: 99%

“…Although the background of this difference is not completely understood, it seems that the culture medium in which embryos are grown has dramatic effect(s) on developmental competence and cryo-withstand of the resultant embryos [6]. In this regard, a number of different embryo culture protocols have been proposed in order to maximize post-warming survival of vitrified embryos.…”

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confidence: 99%

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Antioxidant supplementation of culture medium during embryo development and/or after vitrification-warming; which is the most important?

Hosseini

Forouzanfar

Hajian

et al. 2009

J Assist Reprod Genet

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Purpose To determine the most optimal stage for antioxidant supplementation of culture medium to improve developmental competence, cryotolerance and DNAfragmentation of bovine embryos. Methods Presumptive zygotes were first cultured in presence or absence of β-mercaptoethanol (β-ME), for 8 days. Subsequently, half of the expanded blastocysts developed in both groups were vitrified, warmed within 30 min and post-warming embryos along with their corresponding non-vitrified embryos were cultured for two further days in presence or absence of (100 µM) βME. Results For vitrified and non-vitrified embryos, the best effect was found when βME was added from day 1 of in vitro culture in continuation with post-warming culture period. Day 1-8 supplementation significantly increased the rates of cleavage, day 7 and day 8 blastocyst production. For non-vitrified embryos, βME addition during day 1-8 and/or 9-10 of embryo culture improved both hatching rate and quality of hatched embryos. For vitrified embryos, however, the percentage of DNA-fragmentation (18.5%) was significantly higher (p≤0.05) than that of embryos developed in absence of βME but supplemented with βME during post-warming period (13.5%). Conclusions Exogenous antioxidant increases the chance of embryos, even those of fair-quality, to develop to blastocyst. However, antioxidant inclusion during in vitro embryo development is not sufficient to maintain the redox state of these embryos during the critical period of post-warming embryo culture, and therefore, there should be a surplus source of exogenous antioxidant during post-warming embryo culture.

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Characterization of embryos derived from calf oocytes: Kinetics of cleavage, cell allocation to inner cell mass, and trophectoderm and lipid metabolism

et al. 2000

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Embryos derived from calf oocytes were compared with adult cow oocyte‐derived embryos (1) by studying the kinetics of embryo development using time‐lapse cinematography (2) by evaluating the ratio between inner cell mass (ICM) and trophectoderm (TE) cells in blastocysts (3) by measuring the triglyceride content of the blastocysts. The rate of calf oocyte‐derived embryos reaching the blastocyst stage was reduced (26 vs. 46% for adult derived embryos). Calf oocyte‐derived embryos preferably arrested their development before the 9‐cell stage. Those that developed into blastocysts had cleaved earlier to reach the 2‐cell or 3‐cell stages than embryos that arrested before the 9‐cell stage. The 9‐cell stage tended to appear later in calf oocyte‐derived embryo that reached the blastocyst stage than in adult‐derived embryos. This difference became significant at the morula stage. Accordingly, the fourth cell cycle duration was longer for calf oocyte‐derived embryos. Day 8 blastocysts from both sources had similar total cell numbers (calf: 89 ± 20; cow: 100 ± 30) and cell distribution between TE and ICM. The triglyceride content of day 7 blastocysts was similar for both sources (64 ± 15 vs. 65 ± 6 ng/embryo, respectively). In conclusion, calf oocyte‐derived embryos are characterized by a higher rate of developmental arrest before the 9‐cell stage and by a longer lag phase preceding the major onset of embryonic genome expression. These changes might be related to insufficient “capacitation” of the calf oocyte during follicular growth. Despite these differences, modifications in the quality of the resulting blastocysts were not detected. Mol. Reprod. Dev. 57:346–352, 2000. © 2000 Wiley‐Liss, Inc.

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Comparison of two co-culture systems to assess the survival of in vitro produced bovine blastocysts after vitrification

Cited by 41 publications

References 27 publications

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

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

Antioxidant supplementation of culture medium during embryo development and/or after vitrification-warming; which is the most important?

Characterization of embryos derived from calf oocytes: Kinetics of cleavage, cell allocation to inner cell mass, and trophectoderm and lipid metabolism

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