Development of in vitro matured/in vitro fertilized bovine embryos into morulae and blastocysts in defined culture media

Bavister, Barry D.; Rose-Hellekant, Teresa A.; Pinyopummintr, Tanu

doi:10.1016/0093-691x(92)90251-l

Cited by 137 publications

(62 citation statements)

References 48 publications

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“…In the present work, the addition of serum promoted the earlier appearance of Ž blastocysts, which has been reported by other authors Bavister et al, 1992;Bavister, 1995;Carolan et al, 1995;Koji et al, 1997;Thompson . et al, 1998a;Tricoire et al, 1999 .…”

Section: Discussionsupporting

confidence: 74%

Effects of glucose and protein sources on bovine embryo development in vitro

Gómez

Díez

2000

Animal Reproduction Science

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Oviductal factors may be obtained by ultrafiltration of conditioned medium, added to a simple media and used in bovine embryo culture. In this study, we aimed to analyze the development of bovine embryos produced with oviductal factors compared to those cultured in the presence of BSA or serum, the effects of glucose in presence of these protein supplements, and the ability of oviductal factors to support embryo development during the entire culture period. In vitro produced bovine zygotes from slaughterhouse ovaries were cultured in modified-synthetic oviduct Ž . Ž . Ž . Ž . fluid mSOF alone or supplemented with 1 oviductal factors, 2 BSA and 3 FCS. Oviductal factors showed embryotrophic activity, although with blastocyst rates lower than those in BSA and Ž . FCS. Glucose 1.5 mM added at Day 2 of culture did not affect development in the presence of oviductal factors. The number of cells in expanded blastocysts was unaffected by the presence of glucose or any of the protein supplements used. Both BSA and FCS, respectively, improved blastocyst rates of Day 6 embryos produced with oviductal factors. The effect of oviductal factors was masked by the presence of BSA during the entire culture. FCS promoted an earlier appearance of blastocysts. It is concluded that the effect of glucose on in vitro embryo development depends upon the source of protein. Oviductal factors are not an appropriate supplement for embryos beyond Day 6 of culture in SOF, although blastocyst rates of such embryos may be increased by culturing them in the presence of FCS or BSA. q

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Section: Discussionsupporting

confidence: 74%

Effects of glucose and protein sources on bovine embryo development in vitro

Gómez

Díez

2000

Animal Reproduction Science

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“…This supports well the observations that addition of either pyruvate or lactate greatly facilitates development of early bovine embryos (Kim et al, 1993a, b;Matsuyama et al, 1993;Takahashi and First, 1993 (Thompson et al, 1993). This is further supported by the evidence that glutamine was oxidized by bovine embryos in much the same developmental pattern as pyruvate (Reiger et al, 1992 (Bavister et al, 1992). The rates of development after culture reported here are similar to those reported elsewhere using these culture techniques (Thompson et al, 1992).…”

Section: Introductionsupporting

confidence: 80%

Oxygen uptake and carbohydrate metabolism by in vitro derived bovine embryos

Thompson¹,

Partridge²,

Houghton³

et al. 1996

Reproduction

294

175

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The consumption of oxygen, uptake of pyruvate and glucose and production of lactate were determined for groups of bovine embryos produced in vitro from the one-cell to the blastocyst stage (day 0\p=n-\6of culture). Measurements were made in Hepes-buffered synthetic oviduct fluid medium supplemented with 1.0 mmol pyruvate l \ m=-\ 1, 10 mmol d,l-lactate l \m=-\1 and 1.5 mmol glucose l \m=-\1and also 3 mg BSA ml \m=-\1and, from day 5 of development, 10% (v/v) fetal calf serum. The amount of ATP production was determined from oxygen consumption and the proportion of glucose taken up that could be accounted for by lactate production. The data revealed that oxygen consumption was relatively constant from days 0\p=n-\4of culture (0.24\p=n-\0.27 nl per embryo h \ m=-\ 1) , but increased with the initiation of compaction (0.39 nl per embryo h\m=-\1) and continued to increase with the formation and expansion of the blastocoel (0.9 nl per embryo h \m=-\1). Both pyruvate and glucose uptake followed similar patterns. Furthermore, when plotted against oxygen consumption, both pyruvate and glucose uptake increased significantly (P < 0.001) in a linear relationship (R2 = 0.61 and 0.49, respectively). Lactate production also increased with development and accounted for 40% of glucose uptake at day 0 of culture (putative zygotes), increasing to 70% by day 2 (eight-cell stage) and 100% of glucose uptake from day 4 of culture onwards. ATP production followed a similar pattern to that of oxygen consumption (60\p=n-\85 pmol per embryo h \ m=-\ 1 from day 0 to day 4) increasing with compaction (124 pmol per embryo h \ m=-\ 1) and blastulation (221 pmol per embryo h\m=-\1). For precompaction stages, 93\p=n-\96%of ATP production was derived from oxidative phosphorylation, decreasing to 82% with compaction. ATP produced by oxidative phosphorylation could be accounted for by the uptake of pyruvate, suggesting that bovine embryos produced in vitro utilize little endogenous substrates when appropriate exogenous substrates are present in the culture medium. The data revealed that bovine embryos were dependent on oxidative phosphorylation for energy (ATP) production at all stages of pre-elongation development, with perhaps a shift in dependence towards glycolysis in conjunction with compaction. It follows that oxidizable substrates, such as pyruvate and certain amino acids, are preferred in embryo culture medium during development in vitro.

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“…The cinematographic chamber, gas and thermoregulating systems were as described by Grisart et al (1994) (Dagnelie, 1986 (Bavister et al, 1992;Lim et al, 1994;Pinyopummintr and Bavister, 1994;Rorie et al, 1994;Carolan et al, 1995 (Walker et al, 1992;Bavister, 1995;Carolan et al, 1995). This faster development might explain why blastocysts at day 8 produced in mSOF containing serum contained more cells than did those obtained under serum-free culture conditions.…”

Section: Time-lapse Cinematographic Equipmentmentioning

confidence: 99%

“…Most of these media contain serum or serum albumin as the source of protein. Several laboratories have demonstrated a favourable effect of serum on the devel¬ opment of bovine embryos in defined and BSA-containing media (Bavister et al, 1992;Lim et al, 1994;Rorie et al, 1994;Carolan et al, 1995;Lee and Fukui, 1995). Pinyopummintr and Bavister (1991) emphasized a biphasic effect of bovine calf serum (BCS): inhibiting the first cleavages and promoting further morula and blastocyst development.…”

Section: Introductionmentioning

confidence: 99%

“…Development of bovine embryos beyond the 8-16-cell stage can be achieved using a wide variety of in vitro culture procedures: co-culture with various cell types (for review: see Thibodeaux and Godke, 1992), media conditioned by somatic cells (Eyestone et al, 1991;Kobayashi et al, 1992; Vansteenbrugge et al, 1994) or defined media supplemented or not supplemented with proteins (Bavister et al, 1992; Takahashi and First, 1992; Eckert and Niemann, 1995; Keskintepe et al, 1995). Most of these media contain serum or serum albumin as the source of protein.…”

Section: Introductionmentioning

confidence: 99%

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