Cryopreservation of goat oocytes and in vivo derived 2- to 4-cell embryos using the cryoloop (CLV) and solid-surface vitrification (SSV) methods

Begin, I.; Bhatia, B.; Baldassarre, Hernán; Dinnyés, András; Keefer, Carol L.

doi:10.1016/s0093-691x(02)01257-8

Cited by 99 publications

(48 citation statements)

References 29 publications

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“…The cleavage and development of NT embryos were observed on Day 2 and Days 7-9 (Day 0 = day of fusion). At the end of the culture period, the NT embryos were fixed and stained with Hoechst 33342 by the method of Begin et al [2]. The numbers of nuclei were counted under ultraviolet light.…”

Section: Somatic Cell Nuclear Transfer and Activationmentioning

confidence: 99%

The Effect of Activation Protocols on the Development of Cloned Goat Embryos

Apimeteetumrong

Thuangsanthia²,

Leingcharoen³

et al. 2004

The Journal of Veterinary Medical Science

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ABSTRACT. This study was conducted to compare the developmental competence of somatic nuclear transfer (NT) embryos, after either ionomycin or ethanol activation, in locally bred goats. Donor cells were prepared from the ear skin fibroblasts of a female goat. Cells, at passage 3-8, starved by culturing in 0.5% FCS for 4-8 d, were used for NT. Immature oocytes were obtained from FSH-stimulated goats and matured for 22 hr before enucleation and NT. After fusion, the reconstructed embryos were activated with either ionomycin or ethanol followed by culturing in 6-dimethylaminopurine (6-DMAP) and cytochalasin B (CB), for 3 hr. In experiment I, the fuse d NT embryos (n=63, ionomycin and n=68, ethanol treatments, respectively) were cultured in B 2 with a Vero co-culture system and their developmental competence was evaluated through to Day 9. In experiment II, the NT embryos at the 2-4 cell stage on Day 2 derived from each treatment (ionomycin n=46, and ethanol n=37), were transferred into 10 synchronous recipients. There were no significant differences between the NT embryos derived from the ionomycin and ethanol groups, in fusion (86.3% versus 82.9%), cleavage (90.5% versus 82.4%) and for morula/blastocyst development rates (9.5% versus 5.9%). Sixty percent (3/5) of the recipients from ionomycin became pregnant by midterm (2.5 mts) while only 20% (1/5) from ethanol treatment was pregnant by Day 45. The results demonstrate that activation with either ionomycin or ethanol in combination with 6-DMAP-CB treatment does not affect the development of cloned goat embryos. KEY WORDS: activation, developmental competence, goat, nuclear transfer.J. Vet. Med. Sci. 66(12): 1529-1534, 2004 Since the cloned sheep named "Dolly", was produced by transferring nuclei from adult cells into enucleated oocytes [27], there have been many studies carried out to produce other cloned mammals such as mice, cattle, goats, pigs and rabbits [1,5,19,25,26]. The nuclear transfer (NT) technique has been efficiently used for producing transgenic, cloned farm animal offspring. These animals are capable of producing valuable proteins, which could have a marked impact on the pharmaceutical industry [22] [1,4,6,11,32]. There are a few reports on the development of goat oocytes after parthenogenetic activation by treatment with ionomycin and ethanol, both followed by exposure to 6-diethylaminopurine (6-DMAP) [17,18]. Nevertheless, comparison of the development of NT goat embryos with either ionomycin or ethanol, as an activating agent, has not been reported. The objective of this study was to compare the developmental competence in vitro and in vivo of NT goat embryos, after activation, by using either ionomycin or ethanol, both followed by incubation in 6-DMAP-cytochalasin B (CB). MATERIALS AND METHODSUnless otherwise indicated, all chemicals used in this study were obtained from Sigma-Aldrich Company (St. Louis, MO) and the media from Gibco Invitrogen Corporation (Grand Island, NY).Preparation of donor cells: Donor cells were prepared from an ear skin f...

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Section: Somatic Cell Nuclear Transfer and Activationmentioning

confidence: 99%

The Effect of Activation Protocols on the Development of Cloned Goat Embryos

Apimeteetumrong

Thuangsanthia²,

Leingcharoen³

et al. 2004

The Journal of Veterinary Medical Science

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“…Rall and Fahy [1] were the first to report vitrification, a glass-like solidification, by ultra-rapid cooling of mouse embryos, showing an alternative to traditional freezing methods to avoid chilling injury and ice crystal formation. Vitrification has been widely applied to cryopreserve oocytes in other mammalian species including cattle [2], pigs [3], sheep [4], goats [5], horses [6], buffalo [7], cats [8] and humans [9].…”

mentioning

confidence: 99%

A Comparison of Cryotop and Solid Surface Vitrification Methods for the Cryopreservation of In Vitro Matured Bovine Oocytes

Sripunya

Somfai

Inaba

et al. 2010

Journal of Reproduction and Development

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Abstract. The aim of the present study was to compare the efficacies of the cooling systems of the solid surface (SSV) and Cryotop vitrification methods for cryopreservation of bovine oocytes at the metaphase II stage. The effects of vitrification on oocyte viability, in vitro fertilization (IVF), pronucleus formation and subsequent in vitro development were assessed. In vitro matured (IVM) bovine oocytes were subjected to equilibration and vitrification solutions according to the SSV method, and then the oocytes were vitrified either by dropping onto a cold dry metal surface (SSV group) or by plunging into liquid nitrogen on Cryotop sheets (Cryotop group). Warming was conducted according to the SSV method. Some oocytes were subjected to the cryoprotectants and warming regimen without cooling (Solution control group). The live/dead status of oocytes was evaluated by fluorescein diacetate staining. Live oocytes were subjected to IVF, and the resultant embryos were cultured in vitro. The rates of live oocytes were similar among the Fresh control, Solution control, SSV and Cryotop groups. There was no difference in the rates of fertilization, pronuclear formation and monospermy among these groups. The cleavage rates in the SSV and Cryotop groups (41.6 and 53.2%, respectively) were significantly lower than those in the Fresh control and Solution control groups (65.9 and 61.3%, respectively). The blastocyst rates in SSV and Cryotop groups did not differ (10.3 and 12.8%, respectively); however, they were significantly lower than those in the Fresh control and Solution control groups (36.4 and 24.8%, respectively). The inner cell mass, trophectoderm and total cell numbers in blastocysts did not differ significantly among the Fresh control, Solution control, SSV and Cryotop groups. Our results indicate that IVM bovine oocytes could be cryopreserved successfully using the cooling systems of the Cryotop and Solid Surface Vitrification methods with similar efficacy. Key words: Bovine, Cryotop, Oocyte, Solid surface vitrification, Vitrification (J. Reprod. Dev. 56: [176][177][178][179][180][181] 2010) ryopreservation of bovine oocytes is of great importance in the preservation of female genetic resources. Cryopreserved oocytes can be used for in vitro fertilization (IVF), nuclear transfer and genetic manipulation. Rall and Fahy [1] were the first to report vitrification, a glass-like solidification, by ultra-rapid cooling of mouse embryos, showing an alternative to traditional freezing methods to avoid chilling injury and ice crystal formation. Vitrification has been widely applied to cryopreserve oocytes in other mammalian species including cattle [2] In general, oocytes are more susceptible to cooling damage than zygotes because metaphase spindle microtubule integrity is disrupted during cooling and the high concentration of cryoprotectants affects oocytes during equilibration [10]. Also, differences in the membrane structures seem to make oocytes more sensitive to chilling compared with zygotes [11]. Although the tox...

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“…20,21 The cooling rate and warming rate for CS determined in our laboratory are around 1200°C/min and 700°C/min, respectively (unpublished data). Nevertheless, the improvement of cooling/warming rates with all these technologies is limited and a toxic concentration of CPAs is still needed.…”

Section: Introductionmentioning

confidence: 88%

Vitreous Cryopreservation of Human Umbilical Vein Endothelial Cells with Low Concentration of Cryoprotective Agents for Vascular Tissue Engineering

Zheng

Zhao

Panhwar

et al. 2016

Tissue Engineering Part C: Methods

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Cryopreservation of human umbilical vein endothelial cells (HUVECs) is important to tissue engineering applications and the study of the role of endothelial cells in cardiovascular and cerebrovascular diseases. The traditional methods for cryopreservation by vitrification (cooling samples to a cryogenic temperature without apparent freezing) using high concentration of cryoprotective agents (CPAs) and slow freezing are suboptimal due to the severe toxicity of high concentration of CPAs and ice formation-induced cryoinjuries, respectively. In this study, we developed a method to cryopreserve HUVECs by vitrification with low concentration of CPAs. This is achieved by optimizing the CPAs and using highly thermally conductive quartz capillary (QC) to contain samples for vitrification. The latter minimizes the thermal mass to create ultra-fast cooling/warming rates. Our data demonstrate that HUVECs can be vitrified in the QC using 1.4 mol/L ethylene glycol and 1.1 mol/L dimethyl sulfoxide with more than 90% viability. Moreover, this method significantly improves the attachment efficiency of the cryopreserved HUVECs. The attached cells post-cryopreservation proliferate similarly to fresh cells. Therefore, this study may provide an effective vitrification technique to bank HUVECs for vascular tissue engineering and other applications.

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Cryopreservation of goat oocytes and in vivo derived 2- to 4-cell embryos using the cryoloop (CLV) and solid-surface vitrification (SSV) methods

Cited by 99 publications

References 29 publications

The Effect of Activation Protocols on the Development of Cloned Goat Embryos

The Effect of Activation Protocols on the Development of Cloned Goat Embryos

A Comparison of Cryotop and Solid Surface Vitrification Methods for the Cryopreservation of In Vitro Matured Bovine Oocytes

Vitreous Cryopreservation of Human Umbilical Vein Endothelial Cells with Low Concentration of Cryoprotective Agents for Vascular Tissue Engineering

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