Conventional Freezing of In Vitro-Produced and Biopsied Bovine Blastocysts in the Presence of a Low Concentration of Glycerol and Sucrose

Tominaga, Keiichiro; Iwaki, Fumiyuki; Hochi, Shinichi

doi:10.1262/jrd.18083

Cited by 11 publications

(11 citation statements)

References 21 publications

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“…The experiments with IVP blastocysts showed a similar post-thaw survival rate for biopsied (92.3%) and intact (96.2%) embryos after slow cooling in 0.7M glycerol plus 0.1M sucrose, respectively (Tominaga et al, 2007). In our study, 95.6% of embryos survived the biopsy, in vitro culture during the sex analysis (up to 3.5 hours) and freezing procedures.…”

Section: Discussionsupporting

confidence: 77%

Conception rate after sex determination and cryopreservation of D7 bovine embryos:

Lopatářová¹,

Čech²,

Krontorád³

et al. 2010

Vet. Med. - Czech

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ABSTRACT:The aim of this study was to evaluate conception rate after sex determination and subsequent freezing of Day 7 (D 7 ) bovine embryos and to compare it with transfer of fresh female embryos. High quality embryos obtained from superovulated donors were biopsied and the embryonic cells (5-10) were analysed by PCR using specific primers for the Y-chromosome determinant. Fresh embryos were blade biopsied (n = 172) and subsequently transferred (ET) ipsilaterally to synchronized recipients. Selected embryos (n = 112) were biopsied by cell aspiration and cryopreserved for later transfer. Sex determination was successfully completed in 91.3% of fresh group embryos (44% female) and in 87.5% of freeze-thawed group embryos (45.9% female). The achieved pregnancy rate was 56.5% after fresh transfer of sexed (female) embryos whereas intact embryos (control 1) implanted in 61.9% recipients. 95.6% (43/45) of embryos survived biopsy and cryopreservation. After transfer of freeze-thawed sexed (female) embryos, pregnancy was established in 48.8% of recipients. A similar conception rate was found in intact embryos (50.7%, control 2). The results clearly demonstrate that the microsurgical technique used and subsequent PCR sex analysis allow the rapid commercial exchange of genetic resources on the basis of fresh or frozen sex-desired embryos in embryo transfer programs.

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Section: Discussionsupporting

confidence: 77%

Conception rate after sex determination and cryopreservation of D7 bovine embryos:

Lopatářová¹,

Čech²,

Krontorád³

et al. 2010

Vet. Med. - Czech

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“…Serum additives or bovine serum albumin (BSA)-based preparations are widely used as supplements for cryopreservation medium, despite the undefined nature of serum composition2021. These additives were reported to protect cell membranes during the freezing process22.…”

Section: Discussionmentioning

confidence: 99%

A simple medium enables bovine embryos to be held for seven days at 4°C

Ideta

Aoyagi

Tsuchiya

et al. 2013

Sci Rep

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Cryopreservation methods using liquid nitrogen (LN2) for gametes and embryos are prevalent in mammalian artificial reproduction. However, the pregnancy rate from frozen embryos has not improved over the past two decades because freeze–thawing causes significant damage. The strict regulation of transportation of LN2 containers by airlines also limits exchange between breeders. In this article, we introduce a medium that enabled bovine embryos to be held for up to 7 days at 4°C. A pregnancy rate of 75% (24/32) was obtained for embryos held for 7 days in this medium and transferred to primed recipients. Its constituents were medium 199, foetal bovine serum, and HEPES for buffering. This technique will enable LN2-free storage and air transportation of embryos provided transplantation to recipients can be completed within 7 days.

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“…In order to increase the developmental ability of in vitro produced and cryopreserved embryos, selection based on the kinetics of embryo development or the modification of conditions during maturation, cultivation and cryopreservation are recommended (Massip et al, 1995;Rizos et al, 2001Rizos et al, , 2003Cho et al, 2002;Imai et al, 2002;Mtango et al, 2003;Nedambale et al, 2004;Tominaga et al, 2007). New strategies based on metabolic manipulation with embryos before their cryopreservation have been described by Seidel (2006).…”

Section: Discussionmentioning

confidence: 99%

In vitro production of embryos from high performance cows and the development of frozen-thawed embryos after transfer: a field study

Machatkova¹,

Hulinska²,

Rečková³

et al. 2008

Vet. Med. - Czech

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AbstrAct:In this field study, embryos were derived from genetically highly valuable cows excluded from breeding due to reproductive disorders. Cows, 5 to 10 years old, of Czech Siemmental, Holstein Dairy and Beef Cattle breeds were used as oocyte donors. Oocytes were obtained either in the growth phase of the first follicular wave from cows with synchronized oestrus or in any other phases of follicular development from cows without oestrus synchronization. The embryos were prepared by a standard protocol described previously. The mean number of usable oocytes, transferable and freezable embryos per donor, and the mean percentage of usable, transferable and freezable embryos were assessed. The results were analyzed by Student's-t and Chi-squared tests. The embryos were frozen according to a slow freezing protocol. After thawing, they were transferred to recipients on Day 7 after oestrus. Irrespective of the breed, the mean numbers of usable oocytes and transferable and freezable embryos collected per donor were significantly higher (P < 0.01) for the synchronized than for the nonsynchronized donors (20.4 vs 11.7, 4.3 vs 1.0 and 3.2 vs 0.8, respectively). Similarly, the mean percentages of usable oocytes, transferable and freezable embryos were significantly higher (P < 0.01) for the synchronized than for the nonsynchronized donors (28.5% vs 20.5%, 20.9% vs 9.0% and 15.8% vs 6.5%, respectively). On comparison of the synchronized and nonsynchronized donors of each breed, the difference in the mean percentage of usable oocytes was significant (P < 0.01) in cows of all three breeds, the difference in the mean percentage of transferable embryos was significant in Czech Siemmental and Holstein Dairy cows (P < 0.01) and the difference in the mean percentage of freezable embryos was significant only in Holstein Dairy cows (P < 0.01). After the transfer of 41 frozen-thawed embryos and 43 fresh embryos, 20 heifers and 24 heifers became pregnant, respectively. In conclusion: (a) higher number of oocytes from infertile, genetically valuable cows was recovered in the growth phase compared with the other phases of follicular development; (b) greater development of these oocytes resulted in more embryos for transfer and cryopreservation; (c) the transfer of frozen-thawed and fresh embryos resulted in pregnancy rates of 48.8% and 55.8% , respectively.

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Conventional Freezing of In Vitro-Produced and Biopsied Bovine Blastocysts in the Presence of a Low Concentration of Glycerol and Sucrose

Cited by 11 publications

References 21 publications

Conception rate after sex determination and cryopreservation of D7 bovine embryos:

Conception rate after sex determination and cryopreservation of D7 bovine embryos:

A simple medium enables bovine embryos to be held for seven days at 4°C

In vitro production of embryos from high performance cows and the development of frozen-thawed embryos after transfer: a field study

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