In Vitro Fertilization and Development of Bovine Oocytes Matured in Serum-Free Medium1

Saeki, Kazuhiro; Hoshi, Masaki; Leibfried-Rutledge, M.L.; First, N. L.

doi:10.1095/biolreprod44.2.256

Cited by 106 publications

(45 citation statements)

References 22 publications

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“…Meanwhile, both progesterone and estradiol production reached the highest values when LH was included at 100 ng/ml in the maturation medium, therefore further increase in the level of inclusion of LH resulted in no corresponding increase in progesterone and estradiol production. The results obtained in this study is in line with Zuelke andBrackett, 1990 andSaeki et al, 1991. In conclusion, there is necessity for supplementation of hormones into culture medium for successful bovine in vitro maturation and it is suggested further that studies using abattoir ovaries may not be truly representative of the potential of the bovine ovary because mostly aged and underfed animals are slaughtered.…”

Section: Discussionsupporting

confidence: 92%

In vitro maturation of bovine granulosa cells for steroid hormone production

et al. 2011

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SUMMARYThis study aimed to evaluate the effects of preincubated granulosa cells on in vitro maturation and steroid hormones production in cows. Granulosa cells from the ovaries were cultured using the modified M-199 with Earle's salts. 1% bovine serum albumin was added and 5% fetal calf serum. Also 0.8% of antibiotics was also included. Cultures were incubated at 37°C in a 95% air and 100% relative humidity. There were significant differences among values of progesterone and estradiol concentration obtained for levels of inclusion of FSH and LH. It can be concluded that there is necessity for supplementation of hormones in culture medium for bovine IVM. RESUMENSe evaluó el efecto de células de granulosa preincubadas sobre la maduración in vitro y producción de hormonas esteroides en vacas. Las células de granulosa de los ovarios fueron cultivadas empleando el M-199 modificado con sales de Earle. Se añadió 1% de albúmina de suero bovino y 5% de suero fetal de ternero. También se incluyó un 0,8% de antibióticos. Los cultivos fueron incubados a 37°C con 95% de aire y 100% de humedad relativa. Se registraron diferencias significativas entre las concentraciones de progesterona y estradiol obtenidas con los niveles de inclsuión de FSH y LH. Puede concluirse que es necesario suplementar con hormonas el medio de cultivo para IVM en bovinos.

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Section: Discussionsupporting

confidence: 92%

In vitro maturation of bovine granulosa cells for steroid hormone production

et al. 2011

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“…In the current study, the percentages of oocytes that extruded the first PB and that reached the metaphase II stage, and the percentage of TUNEL-positive cumulus cells were not different between COCs that were matured in serum-free medium or medium supplemented with 10% serum. Other studies have reported that blastocysts are successfully produced from oocytes matured in serum-free medium [42][43][44]. However, blastocysts derived from oocytes matured under serum-free conditions display an increased incidence of apoptosis, which is reversed by the addition of leptin to the serum-free maturation medium [5].…”

Section: Discussionmentioning

confidence: 99%

Leptin Promotes Meiotic Progression and Developmental Capacity of Bovine Oocytes Via Cumulus Cell-Independent and -Dependent Mechanisms1

Paula-Lopes

Boelhauve

Habermann

et al. 2007

Biology of Reproduction

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Leptin has been shown to exert positive effects during the maturation of bovine oocytes, influencing blastocyst development, apoptosis, and the transcript levels of developmentally important genes. The present study was conducted to characterize further the mechanisms of leptin action on oocytes and the role of cumulus cells (CCs) in this context. In the first series of experiments, cumulus-oocyte complexes (COCs) were matured in serum-free medium that contained 0, 1 or 10 ng/ml leptin or in medium that was supplemented with 10% (v/v) estrus cow serum (ECS). Leptin concentrations of 1 and 10 ng/ml stimulated the meiotic progression of oocytes. Moreover, TUNEL staining demonstrated that these leptin doses reduced the proportion of apoptotic CCs. In the second series of experiments, COCs or denuded oocytes (DOs) were matured in the presence of 0 or 10 ng/ml leptin. The percentages of COCs and DOs with extruded polar bodies were increased by leptin. In contrast, positive effects of leptin on fertilization rates and blastocyst development were only observed after treatment of COCs but not of DOs. Leptin treatment of COCs consistently enhanced blastocyst development even after parthenogenetic activation of oocytes or after the removal of CCs before fertilization. The proportion of polyspermic oocytes was not affected by leptin treatment or oocyte denudation. In the third series of experiments, COCs were matured in the presence of 0, 1 or 10 ng/ml leptin. The transcript levels of specific genes were determined by reverse transcriptase-quantitative PCR (RT-qPCR) analysis of cumulus cells and single oocytes. Leptin treatment increased the levels of FAS, FASLG, and STAT3 transcripts in oocytes, but did not affect the LEPR, BAX, and BIRC4 mRNA concentrations. In cumulus cells, leptin treatment increased the mRNA levels for LEPR, STAT3, BAX, BIRC4, and FAS, but did not alter FASLG mRNA abundance. In conclusion, leptin differentially regulates gene expression in oocytes and cumulus cells. Moreover, leptin enhances both oocyte maturation and developmental capacity via cumulus cell-independent and -dependent mechanisms.

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“…While the reasons for such discrepancies are not always clear they are probably partly related to differences in the presence/absence of serum and also due to differences in post-fertilization culture systems. The ability of proteinfree defined culture medium to support hormonally enhanced bovine IVM has been demonstrated (Larson and Parks, 1990;Zuelke and Brackett, 1990;Coskun et al, 1991;Saeki et al, 1991;Ectors et al, 1992).…”

mentioning

confidence: 99%

Development of bovine embryos in vitro following oocyte maturation under defined conditions

Lonergan¹,

Carolan²,

Mermillod³

1994

Reprod. Nutr. Dev.

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Summary -A total of 4 615 immature bovine oocytes were used in a series of experiments aimed at the systematic evaluation of the role of the different components of our in vitro maturation (IVM) medium in imparting developmental competence to the oocytes. The results clearly demonstrate that both tissue culture medium 199 (M199) and synthetic oviduct fluid (SOF) are capable of supporting the IVM of bovine oocytes at high rates in the absence of macromolecular supplements, as evidenced by subsequent development to the blastocyst stage (20 and 25%, respectively). However, both were significantly lower than the control (containing 10% fetal calf serum, 5 pg/ml pLH (porcine luteinizing hormone), 1 wg/ml pFSH (porcine follicle-stimulating hormone), and 1 !ig/ml-17p-estradiol, E2) in terms of blastocyst yield. Inclusion of bovine serum albumin (3 mg/ml) was not beneficial and in fact significantly depressed development when added to SOF. It was shown that the advantage of the control conditions over unsupplemented M199 and SOF was entirely attributable to the presence of serum and that in the absence of serum the inclusion of the above hormone cocktail significantly depressed postcleavage development. When used individually, neither LH (50, 500, 5 000 ng/ml) nor FSH (10, 100, 1 000 ng/ml) improved development over M199 alone. In conclusion, we now have a simple maturation system, using a fully defined medium (M199) in which the search for factors improving the cytoplasmic competence acquisition of maturing cattle oocytes will be possible.

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In Vitro Fertilization and Development of Bovine Oocytes Matured in Serum-Free Medium1

Cited by 106 publications

References 22 publications

In vitro maturation of bovine granulosa cells for steroid hormone production

In vitro maturation of bovine granulosa cells for steroid hormone production

Leptin Promotes Meiotic Progression and Developmental Capacity of Bovine Oocytes Via Cumulus Cell-Independent and -Dependent Mechanisms1

Development of bovine embryos in vitro following oocyte maturation under defined conditions

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