Survival of Porcine Delipated Oocytes and Embryos after Cryopreservation by Freezing or Vitrification.

Nagashima, Hiroshi; Cameron, R. D. A.; Kuwayama, Masashige; Young, Mary; Beebe, L.F.S.; Blackshaw, A. W.; Nottle, Mark B.

doi:10.1262/jrd.45.167

Cited by 51 publications

(37 citation statements)

References 41 publications

(55 reference statements)

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“…Partial removal of lipid from porcine embryos (Nagashima et al 1995) and oocytes (Nagashima et al 1999) also reduces sensitivity to chilling and improves development after vitrification. There are abundant cytoplasmic lipids in porcine oocytes, and the amount of triglyceride, the main component of the lipids, is threefold higher than that in cattle and sheep oocytes (McEvoy et al 2000).…”

Section: Discussionmentioning

confidence: 99%

“…The maturation of vitrified immature porcine oocytes can be improved (from 3.7% to 15.0%) by partial removal of cytoplasmic lipids (Park et al 2005). The effect of delipidization on oocyte vitrification has been reported (Nagashima et al 1999), and it was found that about 56% of the oocytes vitrified after delipidization survived, and 6% of oocytes could develop to morula stage after fertilization by subzonal sperm injection.…”

Section: Discussionmentioning

confidence: 99%

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Improved development by Taxol pretreatment after vitrification of in vitro matured porcine oocytes

Shi¹,

Zhu

Zhang³

et al. 2006

Reproduction

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This study was designed to examine the effect of Taxol pretreatment on vitrification of porcine oocytes matured in vitro by an open pulled straw (OPS) method. In the first experiment, the effect of Taxol pretreatment and fluorescein diacetate (FDA) staining on parthenogenetic development of oocytes was evaluated. In the second experiment, viability, microtubule organization and embryo development of oocytes were assessed after oocytes were exposed to vitrification/warming solutions or after vitrification with or without Taxol pretreatment. The results showed that Taxol pretreatment and/or FDA staining did not negatively influence the oocyte's developmental competence after parthenogenetic activation. After being exposed to vitrification/warming solutions, the survival rate (83.3%) of the oocytes was significantly (P < 0.05) reduced as compared with that in the control (100%). Vitrification/warming procedures further reduced the survival rates of oocytes regardless of oocytes being treated with (62.1%) or without (53.8%) Taxol. The proportions of oocytes with normal spindle configuration were significantly reduced after the oocytes were exposed to vitrification/warming solutions (38.5%) or after vitrification with (10.3%) or without (4.1%) Taxol pretreatment as compared with that in control (76.8%). The rates of two-cell-stage (5.6-53.2%) embryos at 48 h and blastocysts (0 -3.8%) at 144 h after activation were significantly reduced after exposure to vitrification/warming solutions or after vitrification as compared with control (90.9% and 26.6% respectively). However, the proportion of vitrified oocytes developed to two-cell stage was significantly higher when oocytes were pretreated with (24.3%) than without (5.6%) Taxol. These results indicate that pretreatment of oocytes with Taxol before vitrification helps to reduce the damage induced by vitrification and is a potential way to improve the development of vitrified porcine oocytes. Reproduction (2006) 131 795-804

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Improved development by Taxol pretreatment after vitrification of in vitro matured porcine oocytes

Shi¹,

Zhu

Zhang³

et al. 2006

Reproduction

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“…Our results on bovine embryos coincide with the ones reported in the pig by Nagashima et al (27), who found significantly improved in vitro survival rates after freezing delipidated embryos. The same group reported successful vitrification of pig embryos after delipidation (24,28). With bovine IVP embryos, Pangestu et al (30) did not obtain survival of delipidated blastocysts after slow freezing.…”

Section: Discussionmentioning

confidence: 99%

Delipidating in vitro-produced bovine zygotes: Effect on further development and consequences for freezability

Díez

Heyman

Bourhis³

et al. 2001

Theriogenology

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To study the effect of partial removal of intracytoplasmatic lipids from bovine zygotes on their in vitro and in vivo survival, presumptive zygotes were delipidated by micromanipulation and cocultured with Veto cells in B2+10% FCS. Blastocyst rates of delipidated (n=960), sham (centrifuged but not delipidated, n=830) and control embryos (n=950) were 42.1, 42.3 and 39.9% respectively (P > 0.05). Day 7 blastocysts derived from delipidated zygotes had a mean of 123.9 + 45.6 nuclei compared to 137.5 + 32.9 for control blastocysts (P > 0.05). The full-term development of delipidated blastocysts after single transfer to recipients was similar to that of control IVF blastocysts (41.2% vs 45.4% respectively). To assess the effect of delipidation on the embryo tolerance to freczing/thawing, delipidated (n=73), control (n=67) and shmn (n=50) Day 7 blastocysts were frozen in 1.36 M glycerol + 0.25 M sucrose in PBS. After thawing, embryos were cocultured for 72 h with Vero cells in B2+10% FCS. Survival rates at 24 h were not significantly different between groups. However, in the delipidated group, the survival rate after 48 h in culture was significantly higher than in the control group (56.2 vs 39.8, P < 0.02), resulting in a higher hatching rate after 3 days in culture (45.2 vs 22.4, P < 0.02). Pregnancy rates for delipidated and control frozen/thawed embryos were respectively 10.5 and 22.2% (P > 0.05). Electron microscopic observations showed much fewer lipid droplets (and smaller) in delipated blastocysts than in controls. Taken together, our data show that delipidation of one cell stage bovine embryos is compatible with their normal development to term and has a beneficial effect on their tolerance to freezing and thawing at the blastocyst stage. This procedure, however, alters the developmental potential of such blastocysts, suggesting that maternally inherited lipid stores interfere with metabolic recovery after thawing.

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“…Nagashima and colleagues showed that the freezing tolerance of porcine oocytes [11] and early-stage embryos [12,13] can be dramatically increased by removing their cytoplasmic lipid droplets using a process called delipation.…”

mentioning

confidence: 99%

Developmental Ability of Porcine In Vitro Matured Oocytes at the Meiosis II Stage After Vitrification

Ogawa

Ueno

Nakayama

et al. 2010

Journal of Reproduction and Development

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Abstract. The aim of the present study was to investigate whether a combination of cytoplasmic lipid removal (delipation) and treatment by a microtubule stabilizer, paclitaxel, would lead to efficient cryopreservation of porcine in vitro matured (IVM) oocytes at the meiosis II (MII) stage. Vitrification and subsequent re-warming and culture of 109 untreated oocytes produced only 9 blastocysts (8.3%). On the other hand, the post-vitrification blastocyst rate was significantly improved (21/113, 18.6%, P<0.05) when oocytes were treated with 1 μM paclitaxel. Oocyte delipation also significantly increased the post-vitrification blastocyst rate compared with the untreated group (15/37, 40.5%, P<0.05). The delipation-and-paclitaxel group exhibited a significantly higher blastocyst rate (34/75, 45.3%, P<0.05) than the paclitaxel group, although it was not significantly higher than that for the delipation group. In transfer experiment, a total of 109 (18.6%) parthenogenetic blastocysts were obtained from 586 oocytes vitrified with the delipation-andpaclitaxel treatment. Transfer of 72 blastocysts to two recipients resulted in 14 (19.4%) somite stage fetuses. In conclusion, we demonstrated for the first time that by removing cytoplasmic lipid droplets from oocytes and performing a microtubule stabilization procedure, vitrified porcine IVM MII-stage oocytes could efficiently develop to the blastocyst stage while retaining the ability to develop into fetuses. Key words: Cryopreservation, Delipation, In vitro matured oocytes, Pig, Vitrification (J. Reprod. Dev. 56: [356][357][358][359][360][361] 2010) ryopreservation techniques for mammalian oocytes and embryos have been used for three main purposes: to preserve the genes of elite livestock animals and increase the efficiency of animal breeding, preserve valuable genetically modified animals and endangered species and use germ cells effectively in assisted reproductive technology. Thus far, embryo cryopreservation as a practical technique has been used in a variety of species, including experimental and livestock animals [1,2]. On the other hand, cryopreservation of oocytes remains impractical compared with embryos, and successful production of progeny from cryopreserved oocytes has been reported in limited species, including rabbits [3] [8] and rats [9]. As with porcine embryos, unfertilized porcine oocytes are highly sensitive to low temperature [10]. Consequently, cryopreserved porcine oocytes have yet to be used to successfully produce viable piglets.One of the reasons for the low-temperature sensitivity of porcine oocytes and embryos is their high intracellular lipid content. Nagashima and colleagues showed that the freezing tolerance of porcine oocytes [11] and early-stage embryos [12,13] can be dramatically increased by removing their cytoplasmic lipid droplets using a process called delipation.It is known that the meiotic spindle of the oocyte is extremely cryosensitive, and the impaired development of oocytes at meiosis II after cryopreservation has been attribut...

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Survival of Porcine Delipated Oocytes and Embryos after Cryopreservation by Freezing or Vitrification.

Cited by 51 publications

References 41 publications

Improved development by Taxol pretreatment after vitrification of in vitro matured porcine oocytes

Improved development by Taxol pretreatment after vitrification of in vitro matured porcine oocytes

Delipidating in vitro-produced bovine zygotes: Effect on further development and consequences for freezability

Developmental Ability of Porcine In Vitro Matured Oocytes at the Meiosis II Stage After Vitrification

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