Freezability of porcine blastocysts at different peri-hatching stages

Nagashima, Hiroshi; Yamakawa, Hirohito; Niemann, Heinrich

doi:10.1016/0093-691x(92)90045-s

Cited by 52 publications

(35 citation statements)

References 18 publications

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“…The addition of FCS had no effect on the survival of embryos frozen with glycerol or ethylene glycol under the conditions in the present study. Nagashima et al [5] found that the addition of 15% FCS to Whittingham's M-16 as preculture medium decreased post-thaw survival of porcine cultured peri-hatching blastocysts frozen with 1.5 M glycerol. The major differences between the pre-culture medium used in present study and that used by Nagashima et al were the concentration of FCS (5% vs 15%) and the amount of BSA (1.5% vs 0%).…”

Section: Discussionmentioning

confidence: 99%

“…Collected embryos were placed in modified phosphate buffered saline (PB1) [15] containing 10% (v/v) FCS (Commonwealth Serum Laboratories, VIC, Australia) at 37 C and their developmental stages were determined using a dissecting microscope. Hatched blastocysts at an appropriate embryonic diameter for freezing (less than 300 µm) [5] were selected and frozen within 3 h after collection (Experiment 1). Blastocysts that had not hatched were pre-cultured for 18 h to obtain in vitro pre-cultured peri-hatching blastocysts (Experiment 2).…”

Section: Animals and Embryosmentioning

confidence: 99%

“…Except the delipated early cleavage stage embryos, successful cryopreservation of porcine embryos is confined to blastocysts at peri-hatching stage: expanded blastocyst to small (less than 300 µm) hatched blastocyst [5,6]. This condition, together with low embryonic survival following transfer [7][8][9], severely limits the practical application of this technology.…”

mentioning

confidence: 99%

“…The freezability of porcine blastocysts peaks immediately after hatching and then declines rapidly [5]. This, together with the developmental diversity among litter-mate blastocysts [12,13], makes it difficult to obtain effectually suitable blastocysts in vivo for cryopreservation.…”

mentioning

confidence: 99%

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Freezing of in Vivo Derived and in Vitro Pre-Cultured Porcine Blastocysts: Differences of the Cryoprotective Effect between Glycerol and Ethylene Glycol.

Kashiwazaki¹,

Nagashima²,

Ashman³

et al. 1996

Journal of Reproduction and Development

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Abstract. The present study was conducted to compare glycerol and ethylene glycol as cryoprotectants for porcine in vivo derived and in vitro pre-cultured peri-hatching blastocysts. Embryos were surgically collected from superovulated gilts on Day 6. In vivo derived hatched blastocysts (in vivo HB) were frozen with 1.5 M glycerol (1.5 G) or 1.5 M ethylene glycol (1.5 EG). In vitro pre-cultured peri-hatching blastocysts (in vitro pHB), obtained by culturing Day 6 embryos for 18 h in Whitten's medium supplemented with 1.5% bovine serum albumin and with or without 5% fetal calf serum (FCS), were frozen with 1.5 G or 1.8 EG. All embryos were frozen by a slow cooling method using a programmable freezer. The survival of in vivo HB frozen with 1.5 G (82%) was higher (P<0.01) compared with embryos frozen with 1.5 EG (36%). More blastocysts cultured with FCS developed to the hatched blastocyst stage compared with those cultured without FCS (73% vs 33%, P<0.01) within 18 h. The survival of embryos pre-cultured with or without FCS however was not different following freezing with 1.5 G (23% vs 34%) or 1.8 EG (74% vs 81%). The survival rates for embryos frozen with 1.8 EG were significantly higher (P<0.01) than those for the 1.5 G groups. These results demonstrate that more than 80% of in vivo HB can be successfully frozen with 1.5 G and that similar survival rates for in vitro pHB can be achieved by using 1.8 EG. The study also indicates inherent differences in the post-thaw survival of in vivo derived and in vitro pre-cultured peri-hatching blastocysts frozen with 1.5 G.

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Section: Discussionmentioning

confidence: 99%

Section: Animals and Embryosmentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

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Freezing of in Vivo Derived and in Vitro Pre-Cultured Porcine Blastocysts: Differences of the Cryoprotective Effect between Glycerol and Ethylene Glycol.

Kashiwazaki¹,

Nagashima²,

Ashman³

et al. 1996

Journal of Reproduction and Development

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“…The reason why Ham's F-12 was used in the present study was that the medium originally contained linoleic acid (0.084 mg/l) in it. According to the report by Nagashima et al [17] indicating BSA's cryoprotective effect to porcine embryos, BSA was supplemented to the pre-incubation medium at high concentration (12 mg/ml) in the present study. These considerations for the basal experimental conditions were paid to raise the level of porcine embryo survival after freezing and thawing.…”

Section: Discussionmentioning

confidence: 99%

Effect of Dietary Administration with Linoleic Acid and .ALPHA.-Tocopherol on Freezing Tolerance in Porcine Embryos.

Kojima¹,

Iwasaki

Zeniya³

et al. 1996

Journal of Reproduction and Development

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Timing of the first cleavage post-insemination affects cryosurvival of in vitro-produced bovine blastocysts

Dinnyés¹,

et al. 1999

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The time of the first cleavage of bovine zygotes during in vitro culture can affect the rate of development and cell number of the blastocysts. The aim of this work was to study the effect of the timing of first cleavage on the cryosurvival of the resulting blastocysts. Following standard IVM and IVF, zygotes were cultured in modified synthetic oviduct fluid (SOF), with 10% fetal calf serum (FCS) added 48 hr post insemination, in a humidified atmosphere of 5% CO2, 5% O2 and 90% N2. Embryos which cleaved by 24, 27 30, 33, or 36 hr after insemination (IVF) were harvested and further cultured to the blastocyst stage (day 7 or day 8 post IVF). All developing blastocysts on days 7 and 8 were classified into three groups and were cryopreserved by vitrification. Group A consisted of blastocysts (<150 μm, small blastocysts); group B consisted of expanded or hatching blastocysts (>150 μm, large blastocysts); and group C consisted of morphologically poor quality blastocysts. The vitrification solution consisted of 6.5 M glycerol and 6% bovine serum albumin in PBS (VS3a). Thawed embryos were cultured further and survival was defined as the re‐expansion and maintenance of the blastocoel over 24, 48, and 72 hr, respectively. Overall survival and hatching at 72 hr post‐thawing was higher in blastocysts formed by day 7 than those formed by day 8 (60% vs. 40% survival; 63% vs. 45% hatching). Large blastocysts from day‐7 and day‐8 groups survived significantly better than small or poor quality blastocysts (76% vs. 63% and 31%; 72% vs. 30% and 26%, respectively; P < 0.05). Day‐7 blastocysts from the 27‐ and 30‐hr cleavage groups survived significantly better than those from the 36‐hr group (63% and 66% vs. 25%, P < 0.05). Day‐8 blastocysts from later cleaved (30 hr) zygotes had a higher survival than the 27‐hr cleavage groups (52% vs. 26%, P < 0.05). These results indicate that the day of blastocyst appearance, developmental stage, and timing of the first cleavage post‐insemination can influence the cryosurvival of bovine blastocysts following vitrification. Mol. Reprod. Dev. 53:318–324, 1999. © 1999 Wiley‐Liss, Inc.

show abstract

Freezability of porcine blastocysts at different peri-hatching stages

Cited by 52 publications

References 18 publications

Freezing of in Vivo Derived and in Vitro Pre-Cultured Porcine Blastocysts: Differences of the Cryoprotective Effect between Glycerol and Ethylene Glycol.

Freezing of in Vivo Derived and in Vitro Pre-Cultured Porcine Blastocysts: Differences of the Cryoprotective Effect between Glycerol and Ethylene Glycol.

Effect of Dietary Administration with Linoleic Acid and .ALPHA.-Tocopherol on Freezing Tolerance in Porcine Embryos.

Timing of the first cleavage post-insemination affects cryosurvival of in vitro-produced bovine blastocysts

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