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Slow freezing versus vitrification for the cryopreservation of zebrafish (Danio rerio) ovarian tissue.

Marques, L.S, Fossati, A.A.N., Rodrigues, R.B., Da Rosa, H.T., Izaguirry, A.P., Ramalho, J.B., Moreira, J.C.F., Santos, F.W., Zhang, T. and Streit, D.P., 2019. Slow freezing versus vitrification for the cryopreservation of zebrafish (Danio rerio) ovarian tissue. Scientific Reports, 9 (1), 15353.

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DOI: 10.1038/s41598-019-51696-7

Abstract

The aim of the present study was to compare the efficiency of vitrification and slow freezing techniques for the cryopreservation of zebrafish ovarian tissue containing immature follicles. In Experiment 1, assessment of cell membrane integrity by trypan blue exclusion staining was used to select the best cryoprotectant solution for each cryopreservation method. Primary growth (PG) oocytes showed the best percentage of membrane integrity (63.5 ± 2.99%) when SF4 solution (2 M methanol + 0.1 M trehalose + 10% egg yolk solution) was employed. The vitrification solution, which presented the highest membrane integrity (V2; 1.5 M methanol + 5.5 M Me2SO + 0.5 M sucrose + 10% egg yolk solution) was selected for Experiment 2. Experiment 2 aimed to compare the vitrification and slow freezing techniques in the following parameters: morphology, oxidative stress, mitochondrial activity, and DNA damage. Frozen ovarian tissue showed higher ROS levels and lower mitochondrial activity than vitrified ovarian tissue. Ultrastructural observations of frozen PG oocytes showed rupture of the plasma membrane, loss of intracellular contents and a large number of damaged mitochondria, while vitrified PG oocytes had intact mitochondria and cell plasma membranes. We conclude that vitrification may be more effective than slow freezing for the cryopreservation of zebrafish ovarian tissue.

Item Type:Article
ISSN:2045-2322
Group:Faculty of Science & Technology
ID Code:32999
Deposited By: Unnamed user with email symplectic@symplectic
Deposited On:04 Nov 2019 16:04
Last Modified:04 Nov 2019 16:04

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