Sex of IVM, IVF and IVC Blastocysts Produced from Individual Donor Cows.

Tominaga, Keiichiro; Yoneda, Kan; Utsumi, Kyozo

doi:10.1262/jrd.42.35

Cited by 3 publications

(2 citation statements)

References 21 publications

(27 reference statements)

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“…Great variation in the development rate o f em b ry o s to t h e b la s t o c y s t s t a g e a m o n g individual cows and heifers has been reported [39][40][41]. Therefore, we have examined the effect of oocyte donors on sex ratio of embryos derived from IVM-IVF-IVC, but no relationship between male predominance and the developmental rate of embryos was found [42]. Meanwhile, King et al [43] reported that there was significant difference in the sex ratio of morulae bisected and cultured either in vitro or in vivo for 24-30 h, probably due to that the environmental condition around bisected embryos resulted in a specific loss of female embryos.…”

Section: Factors Affecting Sex Determination Of Ivp Embryosmentioning

confidence: 96%

Cryopreservation and Sexing of In Vivo- and In Vitro-Produced Bovine Embryos for Their Practical Use

Tominaga

2004

Journal of Reproduction and Development

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Abstract. My research awarded includes contributions to cryopreservation and sexing of bovine embryos produced in vitro and in vivo, as follows; (1) In vivo-derived morulae and blastocysts were cryopreserved in the presence of 10% glycerol, and the embryos were transferred into recipients after two-step dilution of glycerol in straw, with a practically acceptable pregnancy rate. (2) The survival rate of 16-cell stage embryos frozen in the medium with ethylene glycol was higher than that with DMSO or 1,2-propanediol. Addition of linoleic acid-albumin to culture medium enhanced the survival rate of post-thaw bovine 16-cell stage in vitro-produced (IVP) embryos. (3) Polarization of cytoplasmic lipid droplets by centrifugation of 2-cell stage embryos was found effective to increase freezing tolerance in 16-cell stage embryos developed from the centrifuged embryos, because blastomeres of 16-cell stage embryos were mostly lipid-free. (4) The usefulness of gel-loading tip (GLTip) as a container for ultra-rapid vitrification was demonstrated in IVP embryos from 2-cell to blastocyst stages, with a higher in vitro survival than the conventional two-step freezing. (5) PCR analysis for sexing of in vivo-derived Day-7 embryos indicated that male embryos developed faster and graded higher than female embryos. But such correlation between genetic sex and embryonic development was not found in IVP embryos obtained from individual cows. (6) Addition of 0.1-1.0% deproteinized hemodialysate product from calf blood to culture medium increased the producing efficiency of demi-embryos with good quality. Female embryos rather than male embryos required a longer time to repair after bisection. (7) In vivo-derived bovine embryos after biopsy for sexing by PCR analysis and subsequent vitrification using GL-Tips are available to practical use in the field. (8) Introduction of primer extension preamplification-PCR and purification of DNA product before standard sexing PCR of biopsy samples from Day 3-4 in vitro-derived embryos allowed accurate sex determination, and Day-7 blastocysts developed from Day 3-4 embryos were cryopreserved by GLTip vitrification without a loss of their viability. Thus the field application of bovine embryo transfer is in part supported by improvements of technologies in embryo cryopreservation and sex predetermination.

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Section: Factors Affecting Sex Determination Of Ivp Embryosmentioning

confidence: 96%

Cryopreservation and Sexing of In Vivo- and In Vitro-Produced Bovine Embryos for Their Practical Use

Tominaga

2004

Journal of Reproduction and Development

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“…We also know that the developmental speed of bovine preimplantation embryos changes during periods of early cleavage stages, such as pronuclear formation in the first CD and genomic shift from the maternal to embryonic genome in the 3-4th CD [10,11,27,28], and during periods of morphological change, such as compact formation and hatching from zona pellucidae [10,27,29]. Characteristic changes in cell division, such as chromosomal abnormalities [29][30][31] and a difference in the developmental process because of the sex of embryos [32,33], were also found after blastocyst formation. Using VIVO embryos collected from superovulated donors, we previously observed developmentally retarded embryos and embryos with degenerated blastomeres, as described in other similar reports [20][21][22].…”

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confidence: 99%

Transition of Cell Numbers in Bovine Preimplantation Embryos: In Vivo Collected and In Vitro Produced Embryos

Ushijima

Akiyama²,

Tajima

2008

Journal of Reproduction and Development

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Abstract. The total cell numbers (TCNs) of bovine embryos collected from superovulated donors (VIVO embryos) were counted 0-9 d after ovulation to quantify the developmental process. Using numerical analysis of embryo development, we also compared the developmental process of VIVO embryos, in vitro-fertilized (IVF) embryos and nuclear transfer (NT) embryos obtained from enucleated oocytes and blastomere nuclei. The TCNs of embryos were measured using the air-dry method. Cleavage divisions (CD) of the embryos were obtained using logarithmic transformation of the TCN. The TCN of the VIVO embryos increased significantly (P<0.001) with time. The relationship between the CD of the VIVO embryos at 0-9 d after ovulation and age in days was described by a linear equation with a high correlation (y=1.03x+0. 16, r=0.99), showing that CD occurs about once each day for all blastomeres. However, compared to the VIVO embryos, the TCN of the IVF embryos did not increase from 3-4 d nor after 7 d; the TCN of the NT embryos did not increase after 7 d (P>0.05). The results suggest a delay in development at these developmental stages. The slopes of regression lines of the IVF and NT embryos were significantly (P<0.001) smaller, indicating that quantification of the developmental process of VIVO embryos according to TCN and CD would be useful as criteria for numerical evaluation of the developmental process of bovine in vitro produced embryos. Key words: Bovine, Cleavage division, Preimplantation embryo, Total cell numbers (J. Reprod. Dev. 54: [239][240][241][242][243] 2008) VP embryos such as in vitro maturation/in vitro fertilization/in vitro culture (IVM/IVF/IVC) embryos [1, 2] and nuclear transfer (NT) embryos [3,4] have been shown to lower developmental ability [5,6], lower tolerance to low temperatures and freezing [7,8] and lower pregnancy rates [3,9] compared to in vivo-collected (VIVO) embryos. Therefore, improvements of IVP systems and methods of evaluating produced IVP embryos in the livestock industry to produce calves might be reexamined based on the development of VIVO embryos.Because the preimplantation embryo grows by self-replicating its blastomeres [10,11], total cell numbers (TCN) of embryos and cleavage divisions (CD) of embryos, as calculated from TCN, are commonly used to evaluate the developmental ability of IVC embryos [5,8,12]. A regression relationship between CD and embryo age in days also allows expression of the embryonic developmental process as segmental speed of blastomeres. Therefore, this numerical evaluation is useful for comparing the developmental ability and production methods of embryos [13][14][15]. However, bovine VIVO embryos used in TCN measurements are limited to those at a certain period in the late stage of culture [15], with no one yet having obtained measurements for all developmental stages of IVP embryos continuously during the 0-9 d period. Consequently, we consider that establishing an optimal timeline based on bovine VIVO embryos using the TCN and CD of embryos would be useful ...

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In utero and in vitro sex ratio of bovine embryos and calves originating from the left and right ovaries

Hylan¹

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Sex of IVM, IVF and IVC Blastocysts Produced from Individual Donor Cows.

Cited by 3 publications

References 21 publications

Cryopreservation and Sexing of <i>In Vivo</i>- and <i>In Vitro</i>-Produced Bovine Embryos for Their Practical Use

Cryopreservation and Sexing of <i>In Vivo</i>- and <i>In Vitro</i>-Produced Bovine Embryos for Their Practical Use

Transition of Cell Numbers in Bovine Preimplantation Embryos: In Vivo Collected and In Vitro Produced Embryos

In utero and in vitro sex ratio of bovine embryos and calves originating from the left and right ovaries

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