Please use this identifier to cite or link to this item: http://hdl.handle.net/10174/16569

Title: Cryopreservation of in vitro–produced sheep embryos: Effects of different protocols of lipid reduction
Authors: Romão, Ricardo
Marques, CC
Baptista, MC
Barbas, JP
Horta, AEM
Carolino, N
Bettencourt, E
Pereira, RM
Editors: Elsevier
Keywords: Sheep embryo
Lipid reduction
Conjugated linoleic acid isomer
Cytochalasin
Ultracentrifugation
Cryopreservation
Issue Date: 1-Jul-2015
Publisher: Elsevier
Citation: Romão, R., Marques, C. C., Baptista, M. C., Barbas, J. P., Horta, A. E. M., Carolino, N., ... & Pereira, R. M. (2015). Cryopreservation of in vitro–produced sheep embryos: Effects of different protocols of lipid reduction. Theriogenology, 84(1), 118-126.
Abstract: The low survival of sheep in vitro–produced (IVP) embryos after cryopreservation is a key limiting step to the widespread of this technology. In the present work, different approaches for enhancing cryosurvival of these embryos were compared: embryo delipidation by centrifugation in the absence or presence of cytochalasin D, a cytoskeleton stabilizer or by embryo culture in the presence of different doses of the trans-10 cis-12–conjugated linoleic acid isomer (CLA). Three experiments were conducted. In experiment 1, IVP blastocysts before vitrification were randomly distributed into four groups: control; centrifuged (cent), cytochalasin D (cyto-D), centrifuged + cytochalasin D (cent + cyto-D). In experiment 2, different doses of CLA (25, 50, and 100 μM) were supplemented during embryo culture before vitrification of blastocysts. A control group ran simultaneously. A third experiment was performed to compare both approaches from the previous ones but without the groups with the worst results (groups: control, cyto-D, cent + cyto-D, CLA25, CLA50). In all experiments, embryos integrity and reexpansion were assessed after warming and after 3 hours of culture. In experiment 1, the postwarming integrity rate was the lowest (P < 0.05) in embryos from the cent group (cent: 50.6 ± 10.3% vs. control: 74.6 ± 9.2%, cyto-D: 92.3 ± 9.7%, and cent + cyto-D: 90.5 ± 11.2%), whereas the best (P < 0.05) reexpansion scores were obtained in cent + cyto-D embryos (cent + cyto-D: 2.6 ± 0.28 vs. control: 1.8 ± 0.08, cent: 1.9 ± 0.2, and cyto-D: 1.8 ± 0.31). In experiments 2 and 3, higher (P < 0.05) cleavage rates were observed in CLA25 (50.9 ± 6.2% and 49.2 ± 5.6%, respectively) and CLA50 (48.9 ± 6.2% and 47.6 ± 5.6%, respectively) than those in the control (41.8 ± 6.1% and 40.4 ± 5.4%, respectively) group. In experiment 2, CLA100 presented the lowest (P < 0.002) Day-6 and -7 embryo production rate and quality. After warming, superior (P < 0.02) expansion scores were achieved in CLA25 (3.1 ± 0.29) and CLA50 (3.8 ± 0.17) than in the control (1.9 ± 0.10) group. Similar results were attained in experiment 3. However, although cent + cyto-D embryos showed higher (P = 0.008) postwarming expansion scores than the control (2.8 ± 0.29 vs. 1.9 ± 0.07) group, this score was lower (P = 0.0009) than that in CLA50 embryos (3.8 ± 0.17). In conclusion, our results showed that different protocols of lipid reduction can be successfully applied to improve the cryotolerance of IVP sheep embryos.
URI: http://www.sciencedirect.com/science/article/pii/S0093691X15001053
http://hdl.handle.net/10174/16569
Type: article
Appears in Collections:ICAAM - Publicações - Artigos em Revistas Internacionais Com Arbitragem Científica

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