Consequences of <i>In Vitro</i> Culture Conditions on Embryo Development and Quality

Rizos, Dimitrios; Clemente, María; Bermejo-Álvarez, Pablo; Fuente, J. de la; Lonergan, Patrick; Gutiérrez‐Adán, Alfonso

doi:10.1111/j.1439-0531.2008.01230.x

Cited by 183 publications

(140 citation statements)

References 96 publications

(116 reference statements)

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“…This study showed that KLF4 localisation in both the in vitro and in vivo developed rabbit preimplantation embryos was diffused. As a transcription factor, KLF4 is expected to have a nuclear localisation as reported in the Macaque morulae and blastocysts [3]. The diffused localisation observed in the present study raises more questions related to delay in transport even though it possesses a nuclear localisation signal [55].…”

Section: Discussionmentioning

confidence: 55%

“…Insight into these processes could serve to improve the efficiency of assisted reproduction technologies, development of transgenics, and to understand nuclear reprogramming. However, the embryos developed in vivo and in vitro show differences in their developmental potential [1][2][3] resulting from differences in gene expression patterns [4][5][6]. As the genes of mature spermatozoon and oocyte are silent, and remain inactive after fertilisation, maternal transcripts form a major source for mRNA during the initial stages of embryo development; until the zygotic Genome Activation (ZGA) occurs [7,8].…”

Section: Introductionmentioning

confidence: 99%

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Candidate gene expression patterns in rabbit preimplantation embryos developed in vivo and in vitro

Gibence

Brahmasani

Mahesh

et al. 2014

J Assist Reprod Genet

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Purpose The levels and timing of expression of genes like BCLXL, HDAC1 and pluripotency marker genes namely, OCT4, SOX2, NANOG and KLF4 are known to influence preimplantation embryo development. Despite this information, precise understanding of their influence during preimplantation embryo development is lacking. The present study attempts to compare the expression of these genes in the in vivo and in vitro developed preimplantation embryos. Methods The in vivo and in vitro developed rabbit embryos collected at distinct developmental stages namely, pronuclear, 2 cell, 4 cell, 8 cell, 16 cell, Morula and blastocyst were compared at the transcriptional and translational levels using Real Time PCR and immunocytochemical studies respectively. Results The study establishes the altered levels of candidate genes at the transcriptional level and translational level with reference to the zygotic genome activation (ZGA) phase of embryo development in the in vivo and in vitro developed embryos. The expression of OCT4, KLF4, NANOG and SOX2 genes were higher in the in vitro developed embryos whereas and HDAC1 was lower. BCLXL expression had its peak at ZGA in in vivo developed embryos. Protein expression of all the candidate genes was observed in the embryos. BCLXL, KLF4 and NANOG exhibited diffused localisation whereas HDAC1, OCT4, and SOX2 exhibited nuclear localisation. Conclusions This study leads to conclude that BCLXL peak expression at the ZGA phase may be a requirement for embryo development. Further expression of all the candidate genes was influenced by ZGA phase of development at the transcript level, but not at the protein level.

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Section: Discussionmentioning

confidence: 55%

Section: Introductionmentioning

confidence: 99%

Candidate gene expression patterns in rabbit preimplantation embryos developed in vivo and in vitro

Gibence

Brahmasani

Mahesh

et al. 2014

J Assist Reprod Genet

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“…The resistance of the embryo to cryopreservation is one of the most useful parameters allowing an evaluation of the embryo quality [61]. A reliable embryo production system should be coupled with a cryopreservation protocol leading to improved survival rates after thawing/warming both in vitro and in vivo (i.e., pregnancies).…”

Section: Discussionmentioning

confidence: 99%

In vitro and in vivo quality of bovine embryos in vitro produced with sex-sorted sperm

et al. 2012

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In this work we analyzed the effects of three culture systems on developmental ability of bovine embryos in vitro produced with sexed sperm, the survival to vitrification (cryologic vitrification method) of such blastocysts, and their pregnancy rates after embryo transfer to recipients, both as fresh and after vitrification/warming. Finally, we measured the accuracy of the sorting protocol by a polymerase chain reaction-based method to validate the embryo sex at blastocyst stages. We confirmed an individual effect of the bull as well as development rates of embryos produced with sorted sperm lower than embryos with unsorted sperm, independent of the culture system used. The cryoresistance to vitrification of embryos produced with sexed sperm did not differ from that of conventionally produced embryos (re-expansion rates at 24 and 48 h: 74.6% vs. 75.5%, and 64.5% vs. 68.1% for embryos produced with conventional and sorted sperm, respectively; hatching rates at 48 h: 63.55% vs. 55.5% for embryos produced with conventional and sorted sperm, respectively). Finally, no significant differences were found in pregnancy rates after the embryo transfer of fresh and vitrified/warmed blastocysts (52.8% vs. 42.0%, respectively; P Ͼ 0.05). Male and female embryos produced with sorted sperm showed the same quality in terms of developmental ability, cryoresistance, and pregnancy rates after transfer. Our culture system, coupled with the vitrification in fiber plugs, provides good quality sex-known embryos which survive vitrification at similar rates than embryos produced with conventional unsorted sperm; also it produces good pregnancy rates after transfer of sexed embryos both fresh and after vitrification and warming.

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“…In the latter case, a small number of oocytes per female are usually recovered after OPU (approximately three to six oocytes recovered per nonstimulated cow and session; Rizos et al, 2005;Chaubal et al, 2006). IVEP efficiency has increased in recent years, although the quality of these embryos remains lower than in embryos produced in vivo (reviewed by Rizos et al, 2008). In particular, when embryos were cultured individually or in reduced groups, in addition to a lower efficiency of embryo production, the obtained embryos usually presented a lower number of cells per blastocyst (Paria and Dey, 1990;O'Doherty et al, 1997; Nagao et al, 2008: 5 v. 25 embryos).…”

Section: Introductionmentioning

confidence: 99%

Effect of supplementation of different growth factors in embryo culture medium with a small number of bovine embryos on in vitro embryo development and quality

Ahumada

Salvador

Cebrián-Serrano

et al. 2013

Animal

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When embryos are cultured individually or in small groups, blastocyst yield efficiency and quality are usually reduced. The aim of this work was to investigate the effect of supplementation of the embryo culture medium (CM) with several growth factors (GFs) on embryo development and apoptosis rate when a reduced number of embryos were in vitro cultured. Two experimental studies (ES) were carried out. In ES 1, five treatments were tested to study the effect of GF on embryo development: Control (,30 to 50 embryos cultured in 500 ml of CM); Control 5 (Five embryos cultured in 50 ml microdrops of CM), without addition of GF in either of the two control groups; epidermal GF (EGF); IGF-I; and transforming GF-a (TGF-a) (Five embryos were cultured in 50 ml microdrops of CM with 10 ng/ml EGF, 10 ng/ml IGF-I or 10 ng/ml TGF-a, respectively). In ES 2, following the results obtained in ES 1, four different treatments were tested to study their effect on embryo development and quality (number of cells per blastocyst and apoptotic rate): Control; Control 5; EGF, all three similar to ES 1; EGF 1 IGF-I group (five embryos cultured in 50 ml microdrops of CM with 10 ng/ml EGF and 10 ng/ml IGF-I). In both ESs, it was observed that a higher proportion of embryos cultured in larger groups achieved blastocyst stage than embryos cultured in reduced groups (22.6% v. 14.0%, 12.6% and 5.3% for Control v. Control 5, IGF-I, TGF-a groups in ES 1, and 24.9% v. 17.1% and 19.0% for Control v. Control 5 and EGF in ES 2, respectively; P , 0.05), with the exception of embryos cultured in medium supplemented with EGF (18.5%) or with EGF 1 IGF-I (23.5%), in ES 1 and ES 2, respectively. With regard to blastocyst quality, embryos cultured in reduced groups and supplemented with EGF, alone or combined with IGF-I, presented lower apoptosis rates than embryos cultured in reduced groups without GF supplementation (11.6% and 10.5% v. 21.9% for EGF, EGF 1 IGF-I and Control 5 groups, respectively; P , 0.05). The experimental group did not affect the total number of cells per blastocyst. In conclusion, this study showed that supplementation of the CM with EGF and IGF could partially avoid the deleterious effect of in vitro culture of small groups of bovine embryos, increasing blastocyst rates and decreasing apoptosis rates of these blastocysts.Keywords: bovine embryo, reduced groups, in vitro culture, growth factor, apoptosis ImplicationsIn bovine, in vitro embryo production (IVEP) is especially interesting when applying the ovum pick-up technique (OPU). However, a small number of oocytes per female are usually recovered after OPU, and when embryos are cultured individually or in a reduced group blastocyst yield efficiency and quality are generally reduced. The improvement of IVEP efficiency and quality with low numbers of embryos would be of great interest for applying this technique in the dairy industry. IntroductionIn vitro embryo production (IVEP) has numerous applications in various scientific areas such as animal production or biomedicine. IVEP-rela...

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Consequences of In Vitro Culture Conditions on Embryo Development and Quality

Cited by 183 publications

References 96 publications

Candidate gene expression patterns in rabbit preimplantation embryos developed in vivo and in vitro

Candidate gene expression patterns in rabbit preimplantation embryos developed in vivo and in vitro

In vitro and in vivo quality of bovine embryos in vitro produced with sex-sorted sperm

Effect of supplementation of different growth factors in embryo culture medium with a small number of bovine embryos on in vitro embryo development and quality

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