Development of sheep preimplantation embryos in media supplemented with glucose and acetate

Thompson, Jeremy G.; Parton, G; Cruickshank, G.W.; Smith, J. F.; Wales, R. G.

doi:10.1016/0093-691x(89)90322-1

Cited by 18 publications

(8 citation statements)

References 23 publications

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“…These data are in agreement with those reported for bovine (Camous et al, 1984) and equine embryos (Ball et al, 1993) in co-culture with somatic cells and for ovine embryos in semi-defined medium (Thompson et al, 1989). Embryo viability increased progressively from 2-to 4-cell embryos to 9-to 12-cell embryos.…”

Section: Discussionsupporting

confidence: 92%

Effect of embryo developmental stage and culture conditions on number and quality of ovinein vitroproduced blastocysts

García-García

Domínguez

González-Bulnes

et al. 2006

Zygote

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This study evaluated the final output and quality of in vitro produced blastocysts derived from in vivo recovered sheep embryos cultured at various early developmental stages to blastocyst. A total of 270 embryos were recovered from the oviduct, at different days of the early luteal phase, and were classified into three different developmental stages: 2- to 4-cell (n = 93); 5- to 8-cell (n = 92) and 9- to 12-cell (n = 85). The effect of culture conditions was studied, at the same time, by randomly allocating the embryos to one of four groups: three groups of culture with fresh oviduct monolayers (2, 4 and 5 days old) and a fourth group with 2-day monolayers derived from frozen-thawed oviduct cells. Two control groups were established: first, embryos cultured in semi-defined medium (n = 29) and, second, blastocysts obtained in vivo and cryopreserved (n = 43). Influence on blastocyst yield of embryo developmental stage at the start of culture was statistically significant (p < 0.001). Two- to four-cell embryos showed a significantly lower developmental rate (67.7%) than the 5- to 8-cell (83.6%; p < 0.001) and 9- to 12-cell groups (90.5%; p < 0.0001) and lower quality in terms of blastocyst cryotolerance (56.0 vs. 83.7%; p < 0.005). There were no detected effects relating to the age or handling of the monolayer on the embryo developmental rate, but the day of blastocyst appearance was different between embryos cultured on monolayers derived from fresh or frozen-thawed cells (p < 0.0001); the main influence was on the group of 9- to 12-cell embryos (p < 0.0001). Current results confirm the temporal sensitivities of sheep embryos to in vitro culture, regardless of the culture conditions.

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Section: Discussionsupporting

confidence: 92%

Effect of embryo developmental stage and culture conditions on number and quality of ovinein vitroproduced blastocysts

García-García

Domínguez

González-Bulnes

et al. 2006

Zygote

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“…They were collected in 25 mM 4-(2-hydroxyethy1)-1-piperazineethanesulfonic acid(HEPES)-buffered Synthetic Oviduct Fluid medium (Thompson et al 1989), supplemented with 0.3 mg mL-I bovine serum albumin (BSA) (Sigma, St Louis, USA) (pH 7.4) between Day 1.5 and Day 6, to obtain stages of development ranging from 1-cell to blastocyst.…”

Section: Experiments 1: Glucose Utilization During Preimplantation Devmentioning

confidence: 99%

“…One study has reported that development of 1-cell embryos is dependent on the presence of glucose (Betterbed and Wright 1985) and another (Thompson et al 1989) that later-stage embryos (> 16 cells) have no absolute requirement for this substrate when cultured in vitro. It has also been reported that little glucose is incorporated into glycogen pools by sheep embryos from the 2-to 8-cell stage, but that much of the incorporated glucose is found as low molecular weight components, such as metabolic intermediates (Pike and Wales 1979).…”

Section: Introductionmentioning

confidence: 99%

Glucose utilization by sheep embryos derived in vivo and in vitro

Thompson

Simpson²,

Pugh³

et al. 1991

Reprod. Fertil. Dev.

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Embryos were collected from superovulated donors at various intervals from onset of oestrus, ranging from Day 1.5 to Day 6. In addition, blastocysts obtained from the culture of 1-cell embryos collected in vivo or of oocytes matured and fertilized in vitro were used to assess the effects of in vitro manipulation and culture on glucose utilization. Glycolytic activity was determined by the conversion of [5-3H]glucose to 3H2O, and oxidation of glucose was determined by the conversion of [U-14C]glucose to 14CO2. Glucose utilization increases significantly from the 8-cell stage and during compaction and blastulation. Glucose oxidation was at a relatively low level (5-12% of total utilization) compared with glycolysis. No difference was observed between the glycolytic activity of blastocysts derived from in vivo or in vitro sources. However, glucose oxidation was lower (P less than 0.05) in blastocysts derived from the culture of 1-cell embryos or from oocytes matured and fertilized in vitro. Exogenous tricarboxylic acid cycle substrates (i.e. pyruvate and lactate supplied in the medium) affected the level of glucose oxidation.

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“…The serum-free IVM system reported here demonstrated cleavage rates of 67.7% and blastocyst formation of 34.5% (Table 1). Previous reports have shown that sheep blastocyst production rates can range from 15 to 70% following the IVM of COCs with and without the addition of FCS or oestrus sheep serum (Thompson et al 1989, 1995, Gardner et al 1994, Watson et al 1994, Obrien et al 1996, Walker et al 1996, Wang et al 1998, Grazul-Bilska et al 2003. These serum-based culture systems are commonly supplemented with high pharmacological concentrations of 0.5-10 mg/ml LH and FSH.…”

Section: Discussionmentioning

confidence: 99%

Characterisation of the cellular and molecular responses of ovine oocytes and their supporting somatic cells to pre-ovulatory levels of LH and FSH during in vitro maturation

2012

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The response of Graafian follicles to pre-ovulatory surge levels of FSH and LH in vivo triggers the terminal differentiation of granulosa cells and oocyte maturation. In polyovular species, the LH-driven signalling uses the epidermal growth factor (EGF)-like ligands AREG, EREG and BTC to promote oocyte maturation and cumulus expansion. This experimental series used a physiologically relevant ovine in vitro maturation (IVM) system to evaluate the impact of exposure to pre-ovulatory levels (100 ng/ml) of LH and FSH on ovine cumulus cell expression of EGF-like ligands in vitro. The serum-free sheep IVM system supported high levels (91.4%) of gonadotrophin-induced maturation of cumulus-enclosed oocytes and embryo development to the blastocyst stage (34.5%). Results were equivalent to a serumbased IVM system (85.1% IVM, 25.8% blastocyst rate; PO0.05) but were significantly different (P!0.05) to serum-free medium without gonadotrophins (69.5% IVM; 8.0% blastocyst rate). Ovine BTC was cloned and sequenced. Gonadotrophin-induced AREG, EREG, BTC and EGFR expressions were quantified in cumulus and mural granulosa cells during IVM. A rapid induction of AREG expression was apparent in both cell types within 30 min of gonadotrophin exposure in vitro. LHCGR (LHR) was detected in mural cells and FSHR in both cumulus and mural granulosa cells. The data confirm the involvement of AREG and EGFR during gonadotrophin-induced cumulus expansion, oocyte maturation and the acquisition of developmental competence by sheep oocytes matured in vitro.

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Development of sheep preimplantation embryos in media supplemented with glucose and acetate

Cited by 18 publications

References 23 publications

Effect of embryo developmental stage and culture conditions on number and quality of ovinein vitroproduced blastocysts

Effect of embryo developmental stage and culture conditions on number and quality of ovinein vitroproduced blastocysts

Glucose utilization by sheep embryos derived in vivo and in vitro

Characterisation of the cellular and molecular responses of ovine oocytes and their supporting somatic cells to pre-ovulatory levels of LH and FSH during in vitro maturation

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