Easy, quick and cheap technique to cryopreserve Welsh B pony blastocyst

Guignot, Florence; Blard, Thierry; Barrière, Philippe; Gasgogne, Thierry; Gaude, Yvan; Yvon, Jean-Marie; Mermillod, Pascal; Reigner, Fabrice

doi:10.1016/j.jevs.2016.04.022

Cited by 7 publications

(9 citation statements)

References 0 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Two previous reports described the manual puncture of equine embryos prior to their subsequent vitrification. The first used a glass micropipette of an unspecified size to puncture the embryos and although none of them were transferred to recipient mares, the authors reported a 96% (24/25) survival rate following vitrification, warming and maintenance in culture for 24 hours 15 . The second study manually collapsed 15 equine embryos with a mean diameter of 663µm using an 25G hypodermic needle.…”

Section: Discussionmentioning

confidence: 99%

“…The first used a glass micropipette of an unspecified size to puncture the embryos and although none of them were transferred to recipient mares, the authors reported a 96% (24/25) survival rate following vitrification, warming and maintenance in culture for 24 hours. 15 The second study manually collapsed 15 equine embryos with a mean diameter of 663µm using an 25G hypodermic needle. Vitrification and subsequent warming gave a reported 46% (7/15) pregnancy rate following transfer to recipient mares.…”

Section: Ta B L E 2 Pregnancy Rates For Transferred Embryos Grouped On Bases Of Size and Vitrification Methodologymentioning

confidence: 99%

“…10 Three studies have reported on embryo survival following manual puncture of equine embryos. Guignot et al 15 punctured 28 expanded blastocysts measuring 166-777 µm in diameter and their subsequent collapse was achieved by placing them in medium containing increasing sucrose concentrations prior to vitrification in open-pulled straws. Twenty five of the embryos were warmed to give a reported high survival rate of 96% (24/25) after 24 hours in culture.…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Successful vitrification of manually punctured equine embryos

Wilsher

Rigali

Kovacsy

et al. 2021

Equine Veterinary Journal

View full text Add to dashboard Cite

Background: Successful vitrification of equine expanded blastocysts requires collapse of the blastocoele cavity using a micromanipulator-mounted biopsy pipette on an inverted microscope. Such equipment is expensive and requires user skill.Objectives: To develop a manual method of blastocoele collapse prior to vitrification using commercial products. Study design: In vivo experiment.Methods: Seventy-nine Day 7 or 8 embryos were measured and graded. Twenty were vitrified following micromanipulator-assisted puncture and aspiration before being used to validate commercial human vitrification and warming kits containing, respectively, 2-step concentrations of DMSO and ethylene glycol (7.5%-15% v:v) and decreasing concentrations of sucrose. After warming, embryos were transferred to recipient mares. Once validated, the commercial kits were used to vitrify and warm a further 39 embryos which were punctured manually using a microneedle, 2 (5%) were damaged during puncture and excluded; 20 more embryos were vitrified without puncture. Embryos were grouped as follows: non-punctured ≤ 300µm (n = 10) and >300 to ≤560 µm (n = 10), punctured small (>300 to ≤560 µm; n = 17) and large (>560 µm; n = 10) and exposed to the equilibration solution (ES) in the kit for 6min.An additional group of punctured large embryos was exposed to ES for 8min (n = 10).For the initial warming step, embryos were exposed for 1min to the thawing solution at 42°C, before being moved to a dilution solution at room temperature.Results: Vitrified, manually punctured embryos ≤560 µm exposed to ES for 6min resulted in a pregnancy rate of 82% (14/17). Unpunctured embryos ≤300 µm gave an 80% (8/10) pregnancy rate. Larger unpunctured embryos, punctured embryos >560 µm and embryos exposed to ES for 8min gave significantly reduced pregnancy rates.Main limitations: Limited group sizes. Conclusion:High pregnancy rates can be achieved by manually puncturing ≤560 µm equine embryos prior to their vitrification and subsequent warming in commercial media.

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Ta B L E 2 Pregnancy Rates For Transferred Embryos Grouped On Bases Of Size and Vitrification Methodologymentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Successful vitrification of manually punctured equine embryos

Wilsher

Rigali

Kovacsy

et al. 2021

Equine Veterinary Journal

View full text Add to dashboard Cite

show abstract

“…In mares, embryo size is radically influenced by the PO day selected to implement its recovery (Betteridge et al, 1982;Hochi et al, 1995;Eldridge-Panuska et al, 2005;Choi et al, 2010;McCue et al, 2010;Díaz et al, 2016;Guignot et al, 2016;Wilsher et al, 2019). For the most part, if the embryo is collected either during the sixth or sixth and a half PO day, it is typically ≤ 300 µmØ (Iuliano et al, 1985;Freeman et al, 1991;Eldridge-Panuska et al, 2005;Hudson et al, 2006;Choi et al, 2010;Guignot et al, 2015;Pérez-Marín et al, 2017;Pérez-Marín et al, 2018;Silva et al, 2018).…”

Section: The Relationship Between Day Of Recovery and Embryo Sizementioning

confidence: 99%

Colección, vitrificación y transferencia post-calentamiento de embriones equinos producidos in vivo: una revisión de literatura

2022

Abanico Vet

View full text Add to dashboard Cite

Small equine embryos, equal or smaller than three hundred micrometers in diameter (≤ 300 µmØ) are collected before the seventh post ovulation (PO) day. Big embryos, larger than three hundred micrometers (>300 µmØ) are recovered during or after the seventh PO day. No experiments test the type of holding and washing solutions and to what extent they influence the embryo post vitrification survival. Further knowledge is needed about the mixture of cryoprotectants; either penetrating or non penetrating and the effect they exert over the embryos post vitrification survival and pregnancy rate (PVSPR). The type of embryo carrier, the size of the embryo and the volume of the vitrification solution vary between small and large embryos. A high temperature transfer index (TTI) optimizes the embryo PVSPR. In equids, the type of holding and transfer solutions used during the post vitrification step is an area scarcely explored. This study endeavors to provide information to help in the adaptation of vitrification protocols depending on the embryo size. An additional objective is to ease the access to data about the solutions employed before and during the vitrification; as well as the solutions used during the embryo warming and transfer steps.

show abstract

“…In equine embryos ˃ 300 µmØ, it is important to puncture their EC (38) and extract their BL to promote a reduction in size before the vitrification process (39) . The relevance of EC punction and size reduction in embryos ˃ 300 µmØ has been corroborated in mares; since the application of these procedures resulted in an increased PVPR (23,24,31,38,40,41) . Data also obtained in horses; reported before the year 2010, showed that PVPRs from embryos ˃ 300 µmØ were lower than 40 % (15,29) .…”

Section: Embryo Capsule Punction and Blastocoelic Liquid Extraction Bmentioning

confidence: 99%

Tasa de preñez post vitrificación en los embriones de équidos producidos in vivo. Revisión

Urías-Castro

Boeta

2020

Rev. Mex. Cienc. Pecu.

View full text Add to dashboard Cite

Vitrification is a cryo-preservation method often used in embryos obtained from mares or jennies. It consists in the dramatic reduction of temperature to levels close to -196 °C, that allows the cryopreserving solution containing the embryo to pass from liquid to vitreous state. Several improvements to vitrification protocols have made possible to cryo-preserve embryos with different sizes; since during the first decade after the year 2000, only small embryos were successfully vitrified. Embryos collected at the sixth day post ovulation (PO) are usually smaller or equal to 300 micrometers in diameter (≤ 300 µmØ) and can be routinely vitrified following simple protocols; they have a higher post vitrification pregnancy rate (PVPR) when compared to large embryos which have more than 300 micrometers in diameter (˃ 300 µmØ). The high PVPR of embryos ≤ 300 µmØ is due to an embryo capsule (EC) that is not fully developed yet and has a high permeability to cryo-preserving solutions. At present time, embryos collected either the seventh or eighth day PO are ˃ 300 µmØ and are characterized to have a low post vitrification survival; in order to increase their PVPR their EC might be punctured to make it permeable to cryopreserving solutions. Additionally, there are at least two factors that can be manipulated to increase the PVPR of embryos ˃ 300 µmØ; one is to reduce their size by aspiring their blastocoelic liquid (BL), and the other is to induce a high temperature transfer index (TTI) to rapidly reach -196 °C.

show abstract

Easy, quick and cheap technique to cryopreserve Welsh B pony blastocyst

Cited by 7 publications

References 0 publications

Successful vitrification of manually punctured equine embryos

Successful vitrification of manually punctured equine embryos

Colección, vitrificación y transferencia post-calentamiento de embriones equinos producidos in vivo: una revisión de literatura

Tasa de preñez post vitrificación en los embriones de équidos producidos in vivo. Revisión

Contact Info

Product

Resources

About