Analyses of Developmental Ability of (BALB/c*C57BL/6) F1 and ICR Mouse Embryos Fertilized In Vitro and Their Chromosome at the First Cleavage Division.

Yoshizawa, Midori; Takada, Moriatsu; Nakamoto, S.; Muramatsu, Takashi; Okamoto, Akira

doi:10.1262/jrd.39.115

Cited by 18 publications

(17 citation statements)

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“…The chromosomal abnormalities observed in the present study were mainly polyploidy and mixoploidy. During in vitro fertilization in mice [12,13], cattle [14] and swine [15], the high incidences of polyploidy and mixoploidy are the result of polyspermy. It is likely that the polyploidy and mixoploidy noted in the present study were also the result of polyspermy.…”

Section: Discussionmentioning

confidence: 99%

“…Embryos produced by IVP have a high incidence of chromosomal abnormalities, including mixoploidy and polyploidy; these have been linked to a high frequency of polyspermic fertilization [12][13][14]. Cytogenetic analysis of porcine blastocysts produced in vitro has revealed that about 40% of the blastocysts are abnormal [15]; this is higher than the 7.3% reported for in vivo-derived embryos [16].…”

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confidence: 93%

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The Blastocyst Production Rate and Incidence of Chromosomal Abnormalities by Developmental Stage in In Vitro Produced Porcine Embryos

Ullo

Yoshizawa

Komoriya

et al. 2008

Journal of Reproduction and Development

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Abstract. The present study was conducted to determine the relationship between embryonic development speed at different stages (the cleaved stage at 52 h and the blastocyst stage at 6 days post insemination) and incidences of chromosome abnormalities in in vitro produced porcine embryos. Porcine oocytes were collected from 3-6-mm ovarian follicles obtained at a slaughterhouse and matured in modified NCSU-37 medium for 44-46 h. Following in vitro fertilization with a final concentration of 1 × 10 5 sperm/ml for 3 h, all oocytes were cultured in vitro for 52 h. Day-2 (52 h after insemination) embryos were classified according to their cleaved stages into 2-cell, 3-to 4-cell, 5-to 8-cell, and >8-cell stages; these were cultured separately for additional 4 days (Day 6). The resultant Day-6 blastocysts were classified according to the morphological diameter into 3 grades: Grade A, expanded blastocysts; Grade B, expanding blastocysts; and Grade C, early blastocysts. They were then analyzed chromosomally. The 3-to 4-cell and 5-to 8-cell embryos had significantly high blastocyst development rates (46.1 and 36.9%, respectively), and these blastocysts contained significantly more cells (40.2 and 42.4 cells, respectively) than those derived from 2-cell embryos and >8-cell embryos (28.6 and 26.5 cells, respectively). The incidence of chromosomal abnormalities was significantly higher in the blastocysts derived from 2-cell and >8-cell stage embryos than in the blastocysts derived from the other stage embryos. Furthermore, the grade A blastocysts had the lowest incidence of chromosomal abnormalities (35.3%) and contained the most cells (48.7 cells). Porcine in vitro production (IVP) yielded a high blastocyst rate and an excellent embryo quality when 3-to 4-cell and 5-to 8-cell stage embryos were selected on Day 2 after insemination. The same criteria yielded a higher quality of expanded blastocysts based on the stage of embryo development and morphology. Key words: Blastocyst, Chromosomal abnormality, Embryo, In vitro production (IVP), Pig (J. Reprod. Dev. 54: [22][23][24][25][26][27][28][29] 2008) n mammals, the technology of in vitro production (IVP) of embryos, which involves in vitro maturation and fertilization of oocytes and in vitro culture of the resultant embryos, has become important for increasing the number of offspring from selected high-quality animals and for reducing the generation intervals for breed improvement in livestock [1,2]. Moreover, IVP technology has been used in the production of cloned and transgenic animals, which can be used in biomedical research [3][4][5]. In swine, although embryos have been successfully produced using the IVP system, the development rates to the blastocyst stage and to full term after embryo transfer are still low [6][7][8]. The viability of porcine embryos produced by IVP is low compared with that of in vivo-derived porcine embryos. For example, porcine embryos produced by IVP are of poor quality in terms of the number of cells compared with those produced in vivo [9].A...

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Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 93%

The Blastocyst Production Rate and Incidence of Chromosomal Abnormalities by Developmental Stage in In Vitro Produced Porcine Embryos

Ullo

Yoshizawa

Komoriya

et al. 2008

Journal of Reproduction and Development

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“…IVF using RF spermatozoa was conducted according to our previously described methods [17,18]. Electrically fused oocytes and control group oocytes were introduced into insemination media and cultured for 6 h in a 5% CO 2 incubator.…”

Section: In Vitro Fertilization (Ivf)mentioning

confidence: 99%

Cytogenetic Analysis and Developmental Assessment of Mouse Embryos Derived from In Vitro Fertilization of Oocytes Reconstructed by Meiosis-II Chromosome Transplantation

Mitsui

Yoshizawa

2007

Journal of Reproduction and Development

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Abstract. An electrofusion methodology for transferring meiosis-II chromosomes (M-II-t) has not been completely established. The present study compared the use of two temperatures (fusion at 37 C for Group A and 25 C for Group B) during an electrofusion procedure for mouse oocyte M-II-t and investigated the cytogenetic normality and developmental competence of embryos derived from in vitro fertilization using oocytes reconstructed by M-II-t. The M-II-t oocytes were fertilized in vitro and cultured to the blastocyst stage; the resultant embryos were analyzed cytogenetically. Subsequently, chromosomal normality of the resultant embryos at the prometaphase stage of first cleavage division and the integrity of the meiosis-II spindles of the reconstructed oocytes were analyzed. The success rate of electrofusion in Group B was 92.1%, which was significantly different from that in Group A (49.2%) (P<0.05). The fertilization rates (A, 80.7%; B, 77.2%) and development rates (A, 70.9%; B, 65.5%) in the M-II-t groups were significantly lower than those in the control group (95.0 and 92.2%, respectively) (P<0.05). The incidence of chromosomal abnormalities in the Group A embryos (20.5%) at the blastocyst stage was significantly higher than that in the control group embryos (8.5%) (P<0.05), but the incidence of chromosomal abnormalities in Group B (12.5%) was not significantly different compared with the other groups. A temperature of 25 C during the electrofusion procedure for M-IIt resulted in a good fusion rate, good development rate, and efficient production of chromosomally normal blastocysts. Furthermore, the incidence of chromosomal abnormalities in the first cleavage embryos at the prometaphase stage in Group B (9.6%) did not differ significantly from that in the control group (6.6%). The spindle morphology of the M-II-t oocytes in Group B was normal.

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“…While the cytogenetic study of mouse spermatozoa has been extensively performed [9][10][11][12][13], the study of livestock spermatozoa is less developed. A low fertilizing capacity after in vitro maturation and the lipid contents in livestock oocytes are frequently hindrances for chromosomal analysis; as a result, research in this field has been delayed.…”

Section: Introductionmentioning

confidence: 99%

“…Mouse oocytes have not been utilized for chromosomal analysis of livestock spermatozoa, although there have been abundant studies in mice [9][10][11][12][13]. Therefore, the present study was performed to investigate the effect of oocyte volume in mouse oocyte recipients of livestock spermatozoa on the efficiency of chromosomal analysis.…”

Section: Introductionmentioning

confidence: 99%

A novel method for detection of chromosomal integrity in cryopreserved livestock spermatozoa using artificially fused mouse oocytes

Watanabe

Suzuki

Tateno

et al. 2010

J Assist Reprod Genet

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Purpose The aim of the present study was to investigate the effect of mouse oocyte volume on the efficiency of chromosomal analysis in livestock spermatozoa. Methods Oocytes were injected with bull, ram, boar and dog sperm heads, and then fused with enucleated mouse oocytes. Results The increment of oocyte volume increased the rates of morphologically normal oocytes after sperm injection, which induced much higher rates of overall chromosome detection in bull, ram and dog spermatozoa. The recipient oocyte volume did not affect the chromosomal integrity. Furthermore, in bull, the chromosomal integrity detected by fused mouse oocytes was similar to that derived from a homologous system. On the other hand, chromosomal plates of boar spermatozoa could not be detected despite the use of fused oocytes. Conclusion These data indicate that fused mouse oocytes improved the efficiency of chromosome detection in bull, ram and dog spermatozoa.

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Analyses of Developmental Ability of (BALB/c*C57BL/6) F1 and ICR Mouse Embryos Fertilized In Vitro and Their Chromosome at the First Cleavage Division.

Cited by 18 publications

References 28 publications

The Blastocyst Production Rate and Incidence of Chromosomal Abnormalities by Developmental Stage in In Vitro Produced Porcine Embryos

The Blastocyst Production Rate and Incidence of Chromosomal Abnormalities by Developmental Stage in In Vitro Produced Porcine Embryos

Cytogenetic Analysis and Developmental Assessment of Mouse Embryos Derived from In Vitro Fertilization of Oocytes Reconstructed by Meiosis-II Chromosome Transplantation

A novel method for detection of chromosomal integrity in cryopreserved livestock spermatozoa using artificially fused mouse oocytes

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