Development of in-vitro-derived bovine embryos cultured in 5% CO2 in air or in 5% O2, 5% CO2 and 90% N2

Lim, Jeeyoung; Reggio, B.C.; Godke, R.A.; Hansel, William

doi:10.1093/humrep/14.2.458

Cited by 56 publications

(30 citation statements)

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“…Embryo culture at 5% O2 has shown to produce more blastocysts per cycle with higher mean cell numbers compared to 20% O2, where significantly more blastocysts had abnormally lower cell number (<25) [30]. An efficient development to the blastocyst stage has been achieved for human [31] and bovine [32] embryos using a culture protocol of reduced O2 (5%). Subsequent studies [33,34] have confirmed that blastocysts consume two to four times more O2 than cleavage stage embryos.…”

Section: Discussionmentioning

confidence: 99%

In Vitro Development of Mouse Pronuclear Embryos to Blastocysts in Sequential Media With and Without Co-Culture of Autologous Cumulus Cells

Farouk

Vlad

2008

Journal of Reproduction and Development

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Abstract. The objective of this study was to use mouse embryos as a model system to investigate the effect of co-culture of cumulus cells in Sydney IVF sequential media (Cook) on embryo development, based on the hypothesis that feeder cells in co-culture with a sequential medium could work synergistically to further improve in vitro culture conditions for mammalian preimplantation embryos. The culture systems described here were evaluated by the ability to consistently produce high blastocyst formation rates and high cell number per blastocyst. The role of embryo-to-cell contact was assessed by using Transwell inserts with transparent microporous membranes. Pronuclear embryos of ICR mice were cultured to blastocysts in Cook sequential media, with and without mouse primary cultures of cumulus cells, and with or without inserts. Blastocyst formation rates and cell numbers of in vitro developing embryos in the different culture systems were compared to each other, and to in vivo derived blastocysts. Blastocyst formation rates for Cook medium only was 27.8% (without inserts) and 32.9% (with inserts), whereas Cook-Cumulus cells in identical culture systems was significantly higher at 45.8% (without inserts, P<0.05) and 55.6% (with inserts, P<0.05). When the embryos are suspended above the bottom of the well, for Cook medium significantly lower blastocyst formation rates were observed at 4.2% compared to Cook-Cumulus cells at 17.5% (P<0.05). Mean cell numbers of blastocysts obtained in all co-culture systems were significantly higher (P<0.05) compared to those developing in culture medium only. Although the putative mechanism is as yet unexplained, the improved blastocyst formation rates and cell numbers in co-culture when there is direct contact between the embryo and the cell monolayer suggest that the close proximity between the feeder cells and embryos is in part responsible for these effects. Key words: Blastocyst, Co-culture, Mouse, Pronuclear embryo, Sequential culture medium (J. Reprod. Dev. 54: [385][386][387][388][389][390] 2008) t has been suggested that the co-culture system exerts its beneficial effects by negative [1][2][3][4][5] or positive conditioning [4,[6][7][8][9][10][11] of the culture media, and by embryo-to-cell contact. Negative conditioning or detoxification is the ability of the feeder cells to remove deleterious components from the culture medium whereas positive conditioning is the ability of the feeder cells to modify it by releasing some components (embryotrophic factors) into the culture medium [12,13].There have been studies proposing that embryo-to-cell contact is necessary to allow cellular connections between the zona pellucida and feeder cells for cross-transfer of glycoproteins and other important metabolites [12]. In the cumulus oocyte complex, gap junctions have been shown to be present in the regions of contact between the cumulus cell projections and the oolemma, as well as between adjacent cumulus and granulosa cells [14,15]. Nevertheless, whether gap junctions are present ...

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Section: Discussionmentioning

confidence: 99%

In Vitro Development of Mouse Pronuclear Embryos to Blastocysts in Sequential Media With and Without Co-Culture of Autologous Cumulus Cells

Farouk

Vlad

2008

Journal of Reproduction and Development

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“…In vitro culture of preimplantation embryos under various O 2 concentrations has been carried out in several mammalian species, including mice [8,9], sheep [10], cattle [11][12][13], and humans [14], and reduction of the O2 concentration from 20% to 5% during in vitro culture has been shown to enhance the development to the blastocyst stage. On the other hand, it was reported that there was no significant difference in the in vitro developmental rates to the blastocyst stage of porcine in vivo fertilized embryos between 5% and 20% O 2 in NCSU23 medium [31,32].…”

Section: Discussionmentioning

confidence: 99%

“…The O2 concentration in the mammalian oviduct and uterus is about 5% [7], whereas in vitro cultured embryos are usually maintained under 5% CO2 and 95% air, which contains about 20% O 2 . Reduction of the O 2 concentration from 20% to 5% has been shown to enhance embryonic development in mice [8,9], sheep [10], cattle [11][12][13], and humans [14].…”

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confidence: 99%

Effects of Delipidation and Oxygen Concentration on In Vitro Development of Porcine Embryos

Yoneda

Suzuki

Mori

et al. 2004

Journal of Reproduction and Development

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Abstract. The effects of delipidation and the oxygen (O2) concentration in the atmosphere during culture on in vitro development and H2O2 content were investigated in porcine in vivo fertilized embryos and embryos after in vitro maturation and in vitro fertilization (IVM/IVF embryos). There was no significant difference in the developmental rates to the blastocyst stage between the intact and delipidated IVM/IVF embryos. However, the mean number of cells in blastocysts derived from delipidated IVM/IVF embryos (19.8 ± 0.8 cells) was significantly smaller than that from intact embryos (24.2 ± 1.2 cells). Although there were no significant differences in the developmental rates to the blastocyst stage of intact and delipidated IVM/IVF embryos between the cultures under 5% O2 and 20% O2, the developmental rate of intact IVM/IVF embryos cultured under 5% O2 (27.1%) was significantly higher than that of the delipidated embryos cultured under 20% O2 (19.3%). On the other hand, there was no difference in the developmental rate to the blastocyst stage between in vivo fertilized embryos cultured under 5% O2 and 20% O2. Hydrogen peroxide (H2O2), one of the reactive oxygen species (ROS), is thought to cause damage to embryos. The H2O2 content per embryo derived from oocytes cultured under 5% O2 (in vivo fertilized, 58.0 ± 2.5 pixels; IVM/IVF, 79.6 ± 3.2 pixels) was significantly lower than that (in vivo fertilized, 100.2 ± 3.8 pixels; IVM/IVF, 103.9 ± 3.2 pixels) under 20% O2. Furthermore, the level of H2O2 in delipidated IVM/IVF embryos (94.7 ± 3.9 pixels) was significantly lower than that in intact embryos (103.9 ± 3.2 pixels) cultured under 20% O2. The present results indicate that the delipidation of porcine IVM/IVF embryos and reduction of the O2 concentration decreased the H2O2 level rather than the in vitro developmental rate to the blastocyst stage. Key words: Porcine embryo, In vitro development, Oxygen concentration, Delipidation, Hydrogen peroxide (J. Reprod. Dev. 50: [287][288][289][290][291][292][293][294][295] 2004) he in vitro developmental rate to the blastocyst stage of porcine in vitro maturation (IVM)/in vitro fertilization (IVF) embryos is very low compared with that of in vivo fertilized embryos [1][2][3][4]. Many obstacles must be overcome to obtain a high frequency of in vitro development in porcine I V M / I V F e m b r y o s .F o r e x a m p l e , t h e developmental competence of porcine IVM/IVF e m b r y o s i s m a rk e d l y a f f e c t e d b y t h e s a l t composition [1,5], protein supplementation [6], and small quantities of additional substances [5] in the culture medium. In addition, the oxygen (O2) concentration has been shown to be a major factor causing a difference in the developmental rates of porcine IVM/IVF embryos between in vivo and in

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“…Moreover, anaerobic bacteria are able to colonize the uterine lumen (15,16). Embryos are exceedingly sensitive to oxidative damage (17)(18)(19)(20)(21)(22)(23)(24), and blastocysts produced under low O 2 have significantly more inner cell mass cells than those produced under higher O 2 tensions (25). Accordingly, embryologists generally culture embryos under a low rather than high oxygen atmosphere (17)(18)(19)(20)(21)(22).…”

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confidence: 99%

Low O ₂ tensions and the prevention of differentiation of hES cells

Ezashi

Das²,

Roberts³

2005

Proc. Natl. Acad. Sci. U.S.A.

756

600

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Early-stage mammalian embryos develop in a low O2 environment (hypoxia). hES cells, however, are generally cultured under an atmosphere of 21% O2 (normoxia), under which conditions they tend to differentiate spontaneously. Such conditions may not be the most suitable, therefore, for hES cell propagation. Here we have tested two hypotheses. The first hypothesis was that hES cells would grow as well under hypoxic as under normoxic conditions. The second hypothesis was that hypoxic culture would reduce the amount of spontaneous cell differentiation that occurs in hES colonies. Both hypotheses proved to be correct. Cells proliferated as well under 3% and 5% O2 as they did under 21% O2, and growth was only slightly reduced at 1% O2. The appearance of differentiated regions as assessed morphologically, biochemically (by the production of human chorionic gonadotropin and progesterone), and immunohistochemically (by the loss of stage-specific embryonic antigen-4 and Oct-4 and gain of stage-specific embryonic antigen-1 marker expression) was markedly reduced under hypoxic conditions. In addition, hES cell growth under hypoxia provided enhanced formation of embryoid bodies. Hypoxic culture would appear to be necessary to maintain full pluripotency of hES cells.H1 cell ͉ hypoxia ͉ pluripotent ͉ human chorionic gonadotropin ͉ Oct-4 transcription factor

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Development of in-vitro-derived bovine embryos cultured in 5% CO2 in air or in 5% O2, 5% CO2 and 90% N2

Cited by 56 publications

References 47 publications

In Vitro Development of Mouse Pronuclear Embryos to Blastocysts in Sequential Media With and Without Co-Culture of Autologous Cumulus Cells

In Vitro Development of Mouse Pronuclear Embryos to Blastocysts in Sequential Media With and Without Co-Culture of Autologous Cumulus Cells

Effects of Delipidation and Oxygen Concentration on In Vitro Development of Porcine Embryos

Low O ₂ tensions and the prevention of differentiation of hES cells

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Development of in-vitro-derived bovine embryos cultured in 5% CO2 in air or in 5% O2, 5% CO2 and 90% N2

Cited by 56 publications

References 47 publications

In Vitro Development of Mouse Pronuclear Embryos to Blastocysts in Sequential Media With and Without Co-Culture of Autologous Cumulus Cells

In Vitro Development of Mouse Pronuclear Embryos to Blastocysts in Sequential Media With and Without Co-Culture of Autologous Cumulus Cells

Effects of Delipidation and Oxygen Concentration on In Vitro Development of Porcine Embryos

Low O 2 tensions and the prevention of differentiation of hES cells

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Low O ₂ tensions and the prevention of differentiation of hES cells