Fast Cryopreservation of the Mammalian Embryos in Different Developmental Stages by 0.25 mL Straws Vitrification with One Equilibration Step

Ada Cean, Nicolae Păcală, Alexandra Boleman

Abstract


The aim of our study was to test the cryoprotective proprieties of 7 vitrification media, designed in our laboratory, using the 0.25 mL straws vitrification method, with one equilibration step. As biological material we used mouse females, age 2 months superovulated with 5UI PMSG (Pregnant Mare Serum Gonadotropine) and 5 UI hCG (human Corionic Gonadotropine). For freezing we used embryos in three developmental stages: 2 cells, morula and blastocyst. After recovery, the embryos were placed in equilibration media, after 5 minutes, the embryos were introduced in straws, in vitrification media and plugged directly into liquid nitrogen. After vitrification the straws were thawed in water bath at 37°C, the embryos were rehydrated for 5 minutes and then in vitro cultured. The percent of embryos that rehydrated, resumed development and hatched were registered. The best results were obtained with embryos in morula stage that had a hatching rate of 20.83% when MV1 was used for vitrification. None of the embryos in 2 cells and blastocyst stage hatched after thawing and in vitro culture, regardless of the vitrification media used. From the vitrification media tested, the worst results were obtained with MV4 and MV6, none of the embryos reached hatching stage, regardless of the development stage. The vitrification method in 0.25 mL straws, with one equilibration step can be used for cryopreservation of the morula stage embryos, but is ineffective for vitrification of the 2 cells and blastocyst stage embryo. Media VM4 and VM6 are not suited for vitrification in 0.25 mL straws, with one equilibration step, of mouse embryos.


Keywords


mouse embryos; vitrification media; hatching rate; cryoprotectors

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Scientific Papers Animal Science and Biotechnologies publish open access articles under the terms of the Creative Commons Attribution 4.0 Licence (read full legal code).


LUCRARI STIINTIFICE ZOOTEHNIE SI BIOTEHNOLOGII (SCIENTIFIC PAPERS ANIMAL SCIENCE AND BIOTECHNOLOGIES)

ISSN print 1841 - 9364
ISSN online 2344 - 4576
ISSN-L  1841 - 9364
(former ISSN 1221-5287, E-ISSN 1841-9364)

PUBLISHER: AGROPRINT Timisoara, Romania
PAPER ACCESS: Full text articles available for free
FREQUENCY: Semiannual
PUBLICATION LANGUAGE: English
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Banat´s University of Agricultural Sciences and Veterinary Medicine "King Michael I of Romania" from Timisoara
Bioengineering Faculty of Animal Resources
300645, Timişoara, Calea Aradului 119, Romania

E-mail: spasb [at] animalsci-tm.ro
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