Changes to Extender, Cryoprotective Medium, andIn VitroFertilization Improve Zebrafish Sperm Cryopreservation
| Autor: | Jennifer L Matthews, Joy M Murphy, Huiping Yang, Carrie Carmichael, Monte Westerfield, Terrence R. Tiersch, Zoltán M. Varga |
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| Rok vydání: | 2018 |
| Předmět: |
Male
0301 basic medicine medicine.medical_treatment genetic repository ved/biology.organism_classification_rank.species gene banking Sperm cryopreservation Fertilization in Vitro biomedical law.invention Resource center Andrology 03 medical and health sciences Cryoprotective Agents 0302 clinical medicine law Genetic resources Culture Techniques medicine Animals Model organism model organism Zebrafish Cryopreservation In vitro fertilisation biology ved/biology Extender aquatic biology.organism_classification Spermatozoa 030104 developmental biology Reproductive period resource center Sperm Motility Fish Haus Animal Science and Zoology 030217 neurology & neurosurgery Semen Preservation Developmental Biology |
| Zdroj: | Zebrafish |
| ISSN: | 1557-8542 1545-8547 |
| DOI: | 10.1089/zeb.2017.1521 |
| Popis: | Sperm cryopreservation is a highly efficient method for preserving genetic resources. It extends the reproductive period of males and significantly reduces costs normally associated with maintenance of live animal colonies. However, previous zebrafish (Danio rerio) cryopreservation methods have produced variable outcomes and low post-thaw fertilization rates. To improve post-thaw fertilization rates after cryopreservation, we developed a new extender and cryoprotective medium (CPM), introduced quality assessment (QA), determined the optimal cooling rate, and improved the post-thaw in vitro fertilization process. We found that the hypertonic extender E400 preserved motility of sperm held on ice for at least 6 h. We implemented QA by measuring sperm cell densities with a NanoDrop spectrophotometer and sperm motility with computer-assisted sperm analysis (CASA). We developed a CPM, RMMB, which contains raffinose, skim milk, methanol, and bicine buffer. Post-thaw motility indicated that the optimal cooling rate in two types of cryogenic vials was between 10 and 15°C/min. Test thaws from this method produced average motility of 20% ± 13% and an average post-thaw fertilization rate of 68% ± 16%. |
| Databáze: | OpenAIRE |
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