In vitro culture of bovine IVM-IVF embryos: Cooperative interaction among embryos and the role of growth factors

Keefer, Carol L.; Stice, S.L.; Paprocki, Ann Marie; Golueke, Paul J.

doi:10.1016/0093-691x(94)90491-z

Cited by 68 publications

(42 citation statements)

References 19 publications

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“…Moreover, a significant effect of combination EGF 1 IGF-I on both embryo development and apoptotic rate was found, but without increasing the total cell number per blastocyst. Our results are consistent with several studies that did not show significant effects of EGF on total cells per blastocyst (Keefer et al, 1994; or on ICM and trophoblast cells . In contrast, other studies found a beneficial effect of EGF, increasing the number of cells per blastocyst cultured singly (Paria and Dey, 1990) or in groups (Lee and Fukui, 1995;Mtango et al, 2003).…”

Section: Discussionsupporting

confidence: 93%

“…This controversial result was also observed by ; in their third experiment, the EGF group (5 ng/ml) significantly increased the rate of bovine blastocysts from oocytes, but this fact did not happen in their first experiment (20 embryos/50 ml). In this sense, some authors observed no beneficial effects of EGF in the embryo CM on the blastocyst rate either in individual (mouse: O'Neill, 1997, 0.2 to 2000 ng/ ml; bovine: Keefer et al, 1994, 10 ng/ml) or in group culture (bovine: Flood et al, 1993, 10 ng/ml, Shamsuddin, 1994, 10 to 50 ng/ml, Lee and Fukui, 1995, 10 ng/ml). However, other authors observed differences both in single (mouse: Paria and Dey, 1990, 10 ng/ml; bovine: Lim et al, 2007, 10 ng/ml) and group culture (bovine: Lonergan et al, 1996, 100 ng/ml; Mtango et al, 2003, 10 ng/ml;, 5 ng/ml).…”

Section: Discussionmentioning

confidence: 98%

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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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Section: Discussionsupporting

confidence: 93%

Section: Discussionmentioning

confidence: 98%

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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“…However, for commercial purposes such as use in OPU, where it is usually necessary just to keep the immature oocytes from one donor together, a small number of oocytes/embryos can be cultured (approximately three to six oocytes recovered per nonstimulated cow; Rizos et al, 2005;Chaubal et al, 2006). It has been widely reported that in vitro embryo development in bovine and other mammalian species tends to be suppressed in cultures with a single or low number of embryos (in mouse: Paria and Dey, 1990;Canseco et al, 1992;Lane and Gardner, 1992;Kato and Tsunoda, 1994;in bovine: Palma et al, 1992;Ferry et al, 1994;Keefer et al, 1994;Blondin and Sirard, 1995;Carolan et al, 1996;Donnay et al, 1997;O'Doherty et al, 1997;Ward et al, 2000;Pereira et al, 2005;Fujita et al, 2006). It seems that the culture systems developed to culture embryos in groups could be unsuitable and/or incomplete for individual embryo culture, and there must be some kind of limiting factor or condition in single embryo in vitro culture systems which remain still without knowing (Nagao et al, 2008).…”

Section: Introductionmentioning

confidence: 99%

Effect of number of oocytes and embryos on in vitro oocyte maturation, fertilization and embryo development in bovine

Salvador

Cebrián-Serrano

Salamone

et al. 1970

Span. j. agric. res.

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The aim of this study was to identify the in vitro development stage at which the culture of a single or low number (n = 5 or 10) of oocytes/embryos could impair development in comparison with culture in group (n = 50). In the Experiment 1, it was confirmed that single in vitro embryo production yielded lower cleavage and blastocyst rates than in group (49.4 vs. 83.0%; 0% vs. 37.8%, respectively; p < 0.05). In Experiment 2 and 3, it was observed no effect on embryo development of culturing single or low number of oocytes during maturation and fertilization, respectively. In Experiment 4, it was observed a detrimental effect on blastocyst rate when cultured single or low number of embryos during post-fertilization in vitro culture (2.9; 10.2-10.8; 33.2% in single, low number of embryos (5-10), and controlgrouped, respectively; p < 0.05). In Experiment 5, it was observed that the last part of the culture period (day 3 onwards) seemed to be more affected by the low number of embryos placed in culture. In conclusion, post-fertilization culture, especially on days 3 to 7 after fertilization, seems to be the most important stage for embryo development on single and/or low number (5-10) of embryos culture.Additional key words: single/low embryo culture. ResumenEfecto del número de ovocitos y embriones en la maduración, fertilización y desarrollo embrionario in vitro en vacunoEl objetivo de este trabajo fue identificar la etapa del proceso de producción de embriones in vitro en la cual, el cultivo individualizado o con bajo número de oocitos/embriones (n = 5 ó 10) penaliza al desarrollo embrionario respecto del cultivo en grupo (n = 50). En el Experimento 1, se confirmó que la producción individualizada de embriones in vitro resulta en una menor tasa de división embrionaria y de blastocistos que las obtenidas con el cultivo en grupo (49,4 vs. 83,0%; 0% vs. 37,8%, respectivamente; p < 0,05). En los Experimentos 2 y 3, no se observó efecto sobre el desarrollo embrionario del cultivo individualizado o de bajo número de embriones en las etapas de maduración y fertilización, respectivamente. En el Experimento 4, se observó un efecto negativo sobre la tasa de blastocistos durante el cultivo in vitro post-inseminación cuando se cultivaron los embriones individualmente o con bajo número [2,9; 10,2-10,8; 33,2% en el cultivo individual, con bajo número de embriones (5-10), y grupo control (50), respectivamente; p < 0,05]. En el Experimento 5, se observó que en la última parte del cultivo (día 3 post-inseminación en adelante) es donde más parece que se manifiesta el efecto negativo del cultivo con número reducido de embriones. Como conclusión, el cultivo post-inseminación, especialmente entre los días 3 y 7 de cultivo, parece ser la etapa más determinante del proceso de producción de embriones in vitro para el desarrollo embrionario en el cultivo individualizado y/o en número reducido de embriones (5-10).Palabras clave adicionales: cultivo individualizado/bajo número de embriones.

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“…dermal growth factor [6] or by use of modified culture medium as B2-Menezo medium [7]. However, about 70% of bovine embryos derived from in-vitro maturation, in-vitro fertilization and in-vitro culture will arrest their development before blastocyst stage.…”

mentioning

confidence: 99%

Time-Dependent RNA Synthesis in Early Bovine Embryos Derived from In-Vitro Fertilization.

Iwasaki

Wilmut

Campbell

1997

Journal of Reproduction and Development

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Abstract. RNA synthesis in bovine embryos produced in vitro was investigated to clarify changes in transcriptional activity. Overall 1003 follicular oocytes were matured and fertilized in vitro and subsequently cultured with a cumulus cell monolayer in TCM 199 with 5% fetal bovine serum (FBS). Embryos were transferred to TCM199 supplemented with 100 µCi/ml of [5-3 H]uridine at 26, 35, 43, 50, 68 and 116 h post insemination (hpi) and cultured for 8 h at 39 C. Treated embryos were washed in PBS and the zona pellucida weakened with 5% proteinase K followed by fixation in acetic-alcohol. Preparations were air-dried, dipped in K5 emulsion, and exposed at 4 C. After 4.5 days exposure, the preparations were developed in D19 and the density of 3 H-uridine incorporation was analyzed by image analysis (NIH-157 analysis system). 3 H-Uridine incorporation into nuclei was observed in 43% of 2-cell embryos at 26 hpi although the level was low. The change of 3 Huridine incorporation level was not consistent until 4-cell stage and thereafter the level increased in a time-dependent manner but not dependent on cell number. Incorporation of 3 H-uridine into nucleoli was observed from 50 hpi. Twenty-two embryos at 5-cell to morula were incubated with labelled and unlabelled uridine as negative control showed no incorporation of 3 H-uridine. These results show that the transcription of early bovine embryos fertilized in vitro is already active in 2-cell embryos at 26 hpi, the level increases in a time-dependent manner after 5-cell stage and is not dependent on the cell number.

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In vitro culture of bovine IVM-IVF embryos: Cooperative interaction among embryos and the role of growth factors

Cited by 68 publications

References 19 publications

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

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

Effect of number of oocytes and embryos on in vitro oocyte maturation, fertilization and embryo development in bovine

Time-Dependent RNA Synthesis in Early Bovine Embryos Derived from In-Vitro Fertilization.

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