Cell number and incidence of chromosomal anomalies in bovine blastocysts fertilized in vitro followed by culture in vitro or in vivo in rabbit oviducts

Iwasaki, Setsuo; Nakahara, Tsutomu

doi:10.1016/0093-691x(90)90544-4

Cited by 43 publications

(21 citation statements)

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“…The incidence of chromosomal aberrations in the present study (15.6%), was lower than those in previous reports [5,7,8], since the sperm concentration used in the present study was lower than those in the other reports. Kajihara et al [5] have demonstrated chromosome abnormality in bovine blastocysts derived from in-vitro fertilization to be 22%, composed of triploids and hyperdiploids.…”

Section: Discussioncontrasting

confidence: 56%

“…In investigations of chromosomes, with various mitotic inhibitors, many researchers have used short treatment times for bovine embryos fertilized in vitro. King et al [15] have described a method for preparing chromosomes from bovine blastocysts using 200 µg/ ml colchicine for 2 to 9.5 h. They have shown the mitotic indexes to be 14.7-37.5% in morphologically good blastocysts treated for 8.5-9.5 h, and 4.2-14.3% in embryos treated for 2-4 h. Kajihara et al [5] have reported frequencies of bovine embryos having metaphase plates of approximately 80-83%, and the sexing rate of 63.4% in embryos treated with 0.04 µg/ml colcemid for 1.5-3 h. Tokumaru et al [16] have also reported mitotic indexes of about 5-7% in bovine blastocysts treated with 0.04 µg/ml colcemid for 2 h. Avery et al [17] have shown a low mitotic index of 3.5%, in bovine in-vitro fertilized blastocysts treated with 0.01 µg/ml colcemid for 2 h. Iwasaki et al have reported mitotic indexes of approximately 5 to 12% in bovine blastocysts treated for 4 h with a combination of vinblastine sulfate and podophyllotoxin [7], and a mitoic index of approximately 26% in morulae or blastocysts treated for 6 h with 0.4 µg/ ml vinblastine sulfate [8]. If the treatment time is longer, the mitotic index is higher.…”

Section: Discussionmentioning

confidence: 99%

“…However, we believe that the efficient conditions for chromosome preparation in mouse embryos are not applicable to that in bovine embryos, because the conditions of cultivation in vitro of bovine embryos differs from those of mouse embryos. In the present study, we determined the effective concentration and treatment time of vinblastine sulfate to obtain the greatest number of metaphase plates, and the highest mitotic indexes in chromosome any cytogenetic studies for chromosomal anomalies and sex ratios in bovine embryos M fertilized in vivo [1][2][3] or in vitro [4][5][6][7][8] have been reported to date because of the importance of chromosomal anomalies as an adverse factor after transfer to recipients [9]. For the cytogenetic studies of bovine embryos, it is absolutely necessary to accumulate substantial numbers of analyzable metaphase plates.…”

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confidence: 99%

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Required Concentration and Time of Vinblastine Treatment for Chromosome Preparation in Bovine Blastocysts Derived from In Vitro Fertilization.

Yoshizawa

Matsukawa

Matsumoto

et al. 1998

Journal of Reproduction and Development

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Abstract. The required concentration and time of vinblastine sulfate (a mitotic inhibitor) treatment were determined for successful cytogenetic diagnoses, such as normality and sex in bovine blastocysts, derived from in vitro fertilization. Chromosome preparations were made from blastocysts treated for 17 h, in media containing vinblastine sulfate, with various concentrations of 0, 30, 100, 200 and 300 ng/ml. Vinblastine sulfate was effective at a concentration of 100-300 ng/ml, with respect to the average numbers of metaphase plates and the mitotic indexes (% of mitoses) in the embryos. To determine the sufficient treatment time, embryos were placed in a medium containing 100 ng/ml vinblastine sulfate for 10, 17 and 24 h. The required time of vinblastine sulfate treatment was 10 h. The average length of the no.1 chromosome shortened with the prolongation of treatment time but not with significant difference. Most of the embryos (approximately 98%) treated with vinblastine sulfate had metaphase plates, and approximately 75% of them were numerically analyzable. A high incidence of polyploidy and a sex ratio showing male advantage were found in the present study.

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Section: Discussioncontrasting

confidence: 56%

Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

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Required Concentration and Time of Vinblastine Treatment for Chromosome Preparation in Bovine Blastocysts Derived from In Vitro Fertilization.

Yoshizawa

Matsukawa

Matsumoto

et al. 1998

Journal of Reproduction and Development

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“…Early pig blastocysts transferred to the mouse oviduct showed a 2-fold increase in cell division over comparable blastocysts grown in vitro (Ebert & Papaioannou, 1989). A similar tendency was observed in the bovine embryos derived from in-vitro fertilization and transferred to a rabbit oviduct (Iwasaki & Nakahara, 1990). Handyside & Hunter (1984) established the double dye technique for differential inner cell mass (ICM) and trophectoderm cell counts of mouse blastocysts.…”

Section: Introductionmentioning

confidence: 84%

Morphology and proportion of inner cell mass of bovine blastocysts fertilized in vitro and in vivo

Iwasaki¹,

Yoshiba²,

Ushijima³

et al. 1990

Reproduction

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198

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Summary. The morphology and proportion of inner cell mass (ICM) of bovine blastocysts cultured in vitro or in vivo in rabbit oviducts after in-vitro fertilization of in-vitro matured follicular oocytes were compared with those of blastocysts fertilized in vivo by a differential fluorochrome staining technique. The delineation of each ICM cell was improved by the transfer of embryos derived from in-vitro fertilization to a rabbit oviduct although the cell\p=n-\cellcontacts of ICM cells were not as tight as those from in-vivo fertilization. The proportions (15\m=.\8 and 14\m=.\9%) of ICM in blastocysts cultured in vitro at early and expanded stages were significantly lower than those cultured in rabbit oviducts after in-vitro fertilization and fertilized in vivo. These results show that the transfer of bovine embryos derived from in-vitro fertilization to the rabbit oviduct increased the proliferation of ICM cells to the level of embryos fertilized in vivo although the cell\p=n-\cellcontact of ICM cell is not improved by the process.

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“…When embryos were cultured in modified CR1aa, the numbers of cells was not affected either with nor without cumulus. As the number of cells was considered to be one of the qualitative indicators of embryo development [5,12,20], cultures in Groups 1 and 2 are more appropriate than that in Group 3 for IVM-IVF embryos. The difference may be due to containing 5.56 mM glucose in TCM199 and the difference between CR1aa based media and TCM199 in the development rate and speed depending on the composition of the medium.…”

Section: Discussionmentioning

confidence: 99%

Different Factors Affect Developmental Competence and Cryotolerance in in vitro Produced Bovine Embryo.

Imai

Matoba

Dochi

et al. 2002

The Journal of Veterinary Medical Science

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ABSTRACT. The present study was conducted to examine the effects of culture systems and culture media on developmental competence and freezability of bovine embryos obtained by in vitro culture of in vitro matured and fertilized (IVM-IVF) oocytes. No significant difference was observed in the proportions of oocytes developed to blastocysts, the speed at which the oocytes reached the blastocyst stage and the number of cells, when the IVM-IVF oocytes were cultured in CR1aa with or without cumulus cells. Nevertheless, more of the IVM-IVF oocytes cultured either with or without cumulus cells in CR1aa were seen to reach the blastocyst stage much sooner than those cultured with cumulus cells in TCM199 (P<0.05). The proportion of embryos developed to the blastocyst stage by day 7 in CR1aa culture was significantly higher than embryos cultured in TCM199. Viability after frozen-thawed blastocysts were obtained in vitro, was seen in a significantly higher percentage of embryos cultured in TCM199 and developed to the hatched blastocysts than in those cultured in CR1aa (P<0.05). These results indicate that CR1aa was superior to TCM199 for the potential developmental of IVM-IVF oocytes to blastocysts during in vitro culture regardless of co-culture with or without cumulus cells. But the freezability of blastocysts developed in CR1aa was inferior to those developed in TCM199. KEY WORDS: culture medium, developmental competence, freezability, in vitro culture, in vitro produced bovine embryo.J. Vet. Med. Sci. 64(10): 887-891, 2002 In vitro culture of bovine embryos derived from IVM-IVF have succeeded in producing calves [10,21]. An adequate in vitro culture system for bovine zygotes is required for large scale embryo production by IVM-IVF and genetic improvement by means of ovum pick-up and IVM-IVF. This culture system is used for the development and efficient utilization of embryonic techniques in cattle, for example for embryonic or somatic cell nuclear transfer and in transgenic animals. Bovine IVM-IVF zygotes were first cultured with cumulus cells [8,10], oviduct epithelial cells [9,21], granulosa cells [13], amnion cells [2] and buffalo rat liver cells [36] in TCM199 supplemented with bovine serum, but there are many differences between in vitro produced bovine embryos and those produced in vivo. It was reported that in vitro produced embryos have darker cytoplasm and lower density [24,25]. CR1aa medium was developed for in vitro culture of bovine IVM-IVF zygotes [15,29,30], but the optimum culture system for IVM-IVF oocytes and freezability of blastocysts cultured in CR1aa still remain to be clarified. The chilling sensitivity should be considered as a factor affecting the quality of in vitro produced bovine blastocysts.The aim of the present study was to examine the effects of culture systems and culture media on the development competence and the quality of bovine embryos obtained by in vitro culture of IVM-IVF oocytes. MATERIALS AND METHODS Collection of oocytes and in vitro maturation:Bovine ovaries were obtained ...

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Cell number and incidence of chromosomal anomalies in bovine blastocysts fertilized in vitro followed by culture in vitro or in vivo in rabbit oviducts

Cited by 43 publications

References 12 publications

Required Concentration and Time of Vinblastine Treatment for Chromosome Preparation in Bovine Blastocysts Derived from In Vitro Fertilization.

Required Concentration and Time of Vinblastine Treatment for Chromosome Preparation in Bovine Blastocysts Derived from In Vitro Fertilization.

Morphology and proportion of inner cell mass of bovine blastocysts fertilized in vitro and in vivo

Different Factors Affect Developmental Competence and Cryotolerance in in vitro Produced Bovine Embryo.

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