An Attempt of Cryopreservation of Mouse Embryos at the ACTREC Laboratory Animal Facility in India

Thorat, Rahul; Ingle, Arvind

doi:10.1538/expanim.61.139

Cited by 7 publications

(6 citation statements)

References 34 publications

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“…There are some reports of significant differences between strains in the survival rate of cryopreserved embryos using the frozen method [31,34]. In Experiment 2, the survival rates for the * There were no significant differences among any groups (P > 0.01).…”

Section: Discussionmentioning

confidence: 95%

“…Although several methods are available for cryopreservation of early mouse embryos, including freezing [35] and vitrification [28], the survival rate of early embryos after cryopreservation tends to be higher for the vitrification technique with little difference among strains [18] compared to freezing [29][30][31]34]. Embryo development of multiple strains after cryopreservation also tends to be better with vitrification than with freezing [7].…”

Section: Introductionmentioning

confidence: 99%

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Strain preservation of experimental animals: Vitrification of two-cell stage embryos for multiple mouse strains

2015

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Strain preservation of experimental animals is crucial for experimental reproducibility. Maintaining complete animal strains, however, is costly and there is a risk for genetic mutations as well as complete loss due to disasters or illness. Therefore, the development of effective vitrification techniques for cryopreservation of multiple experimental animal strains is important. We examined whether a vitrification method using cryoprotectant solutions, P10 and PEPeS, is suitable for preservation of multiple inbred and outbred mouse strains. First, we investigated whether our vitrification method using cryoprotectant solutions was suitable for two-cell stage mouse embryos. In vitro development of embryos exposed to the cryoprotectant solutions was similar to that of fresh controls. Further, the survival rate of the vitrified embryos was extremely high (98.1%). Next, we collected and vitrified two-cell stage embryos of 14 mouse strains. The average number of embryos obtained from one female was 7.3-33.3. The survival rate of vitrified embryos ranged from 92.8% to 99.1%, with no significant differences among mouse strains. In vivo development did not differ significantly between fresh controls and vitrified embryos of each strain. For strain preservation using cryopreserved embryos, two offspring for inbred lines and one offspring for outbred lines must be produced from two-cell stage embryos collected from one female. The expected number of surviving fetuses obtained from embryos collected from one female of either the inbred or outbred strains ranged from 2.9 to 19.5. The findings of the present study indicated that this vitrification method is suitable for strain preservation of multiple mouse strains.

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Section: Discussionmentioning

confidence: 95%

Section: Introductionmentioning

confidence: 99%

Strain preservation of experimental animals: Vitrification of two-cell stage embryos for multiple mouse strains

2015

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“…In Experiment 1, we vitrified BDF1 strain 2‐cell stage mouse embryos, which are known to have a high cryotolerance . The results clearly showed that a cryopreservation test using embryos with a high cryotolerance cannot manifest potential difference in performance of the methods.…”

Section: Discussionmentioning

confidence: 98%

Hollow fiber vitrification allows cryopreservation of embryos with compromised cryotolerance

Uchikura

Matsunari

Maehara

et al. 2019

Reprod Medicine & Biology

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Purpose This study aims to demonstrate vitrification methods that provide reliable cryopreservation for embryos with compromised cryotolerance. Methods Two‐cell stage mouse embryos and in vitro produced porcine embryos were vitrified using the hollow fiber vitrification (HFV) and Cryotop (CT) methods. The performance of these two methods was compared by the viability of the vitrified‐rewarmed embryos. Results Regardless of the method used, 100% of the mouse 2‐cell embryos developed successfully after vitrification‐rewarming into the blastocyst stage, whereas vitrification tests using porcine morulae with the HFV method produced significantly better results. The developmental rates of vitrified porcine morula into the blastocyst stage, as well as blastocyst cell number, were 90.3% and 112.3 ± 6.9 in the HFV group compared with 63.4% and 89.5 ± 8.1 in the CT group (P < .05). Vitrification tests using 4‐ to 8‐cell porcine embryos resulted in development into the blastocyst stage (45.5%) in the HFV group alone, demonstrating its better efficacy. The HFV method did not impair embryo viability, even after spontaneous rewarming at room temperature for vitrified embryos, which is generally considered a contraindication. Conclusion Vitrification test using embryos with compromised cryotolerance allows for more precise determining of effective cryopreservation methods and devices.

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“…ACTREC laboratory animal facility have mouse embryo cryo repository for in-house usage. W e have already cryopreserved some of our valuable strains (Thorat and Ingle, 2012). For embryo collection, we need to euthanize more numbers of donor females and require significant time and efforts.…”

Section: Safeguard the Valuable Strainsmentioning

confidence: 99%

Tackling the Covid-19 Pandemic in Animal Facilities: ACTREC Perspectives

Thorat

Ingle

2020

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The Novel Coronavirus outbreak gave learning lesson to the entire world especially about how to fight with the invisible enemy. The scientific community is engaged in exploring the pathogenesis of this Novel disease and inventing the antivirals. At the same time, laboratory animal facility key personnel are busy in carrying out facility operations without compromising the laboratory animal welfare and on-going animal research. In general, every laboratory animal facility establishes the disaster management plan considering the possibility of fire, earthquake, cyclones or flood. Advanced Centre for Treatment, Research and Education in Cancer (ACTREC) Animal Facility also has well documented disaster plan in place. However, incorporation of preparedness plan against disaster due to pandemic was never thought of. However, with our extensive experience, great team efforts and timely decisions, we could continue facility operations without compromising animal welfare, on-going animal research and health of animal care personnel. It is the need of hour to make a preparedness plan in advance to respond to any kind of such pandemics in the future. We wish to share our experience gained during SARS-CoV-2 pandemics. This data is concerned about us but may be helpful to other animal facilities in India and/ or across the globe.

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An Attempt of Cryopreservation of Mouse Embryos at the ACTREC Laboratory Animal Facility in India

Cited by 7 publications

References 34 publications

Strain preservation of experimental animals: Vitrification of two-cell stage embryos for multiple mouse strains

Strain preservation of experimental animals: Vitrification of two-cell stage embryos for multiple mouse strains

Hollow fiber vitrification allows cryopreservation of embryos with compromised cryotolerance

Tackling the Covid-19 Pandemic in Animal Facilities: ACTREC Perspectives

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