Slow freezing versus vitrification for the cryopreservation of zebrafish (Danio rerio) ovarian tissue
Authors: 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.
Journal: Scientific Reports
Volume: 9
Issue: 1
eISSN: 2045-2322
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.
https://eprints.bournemouth.ac.uk/32999/
Source: Scopus
Slow freezing versus vitrification for the cryopreservation of zebrafish (Danio rerio) ovarian tissue.
Authors: 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.
Journal: Sci Rep
Volume: 9
Issue: 1
Pages: 15353
eISSN: 2045-2322
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.
https://eprints.bournemouth.ac.uk/32999/
Source: PubMed
Slow freezing versus vitrification for the cryopreservation of zebrafish (<i>Danio rerio</i>) ovarian tissue
Authors: 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 Jr, D.P.
Journal: SCIENTIFIC REPORTS
Volume: 9
ISSN: 2045-2322
DOI: 10.1038/s41598-019-51696-7
https://eprints.bournemouth.ac.uk/32999/
Source: Web of Science (Lite)
Slow freezing versus vitrification for the cryopreservation of zebrafish (Danio rerio) ovarian tissue.
Authors: 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.
Journal: Scientific reports
Volume: 9
Issue: 1
Pages: 15353
eISSN: 2045-2322
ISSN: 2045-2322
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.
https://eprints.bournemouth.ac.uk/32999/
Source: Europe PubMed Central
Slow freezing versus vitrification for the cryopreservation of zebrafish (Danio rerio) ovarian tissue.
Authors: 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.
Journal: Scientific Reports
Volume: 9
Issue: 1
ISSN: 2045-2322
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.
https://eprints.bournemouth.ac.uk/32999/
Source: BURO EPrints