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Investigating sperm cryopreservation in a model tunicate, Ciona intestinalis sp. A
Authors:Gerarda Sorrenti  Anna Bagnoli  Valentina Miraglia  Fabio Crocetta  Valentina Vitiello  Filomena Ristoratore  Paola Cirino  Giovanni Sansone  Paolo Sordino
Institution:1. Stazione Zoologica Anton Dohrn, Villa Comunale, I-80121 Naples, Italy;2. Laboratory of Cryobiology, Department of Biological Sciences, University of Naples “Federico II”, Via Mezzocannone 16, I-80134 Naples, Italy;3. CRIAcq, Interdepartmental Research Center for Acquaculture, University of Naples “Federico II”, Via Università 100, Portici, I-80055 Naples, Italy;4. CNR ISAFOM – Institute for Agricultural and Forest Systems in the Mediterranean, National Research Council, Str. le Lancia, Blocco Palma I, Zona Industriale, I-95121 Catania, Italy
Abstract:In cryopreservation procedures, the capacity to protect the cells from freezing and thawing processes is sensitive to the choice of the cryoprotective agent (CPA) and to its optimal concentration. The advancement of research on Tunicate model species has raised interest in liquid nitrogen cryopreservation for the storage and distribution of genetic resources. Ciona intestinalis (Linnè, 1767) consists of a complex of cryptic taxa that are central to several areas of investigation, from comparative genomics to invasive biology. Here we investigated how five CPAs, three chilling rates and two freezing rates influence semen cryopreservation in C. intestinalis sp. A. By using larval morphology and motility as endpoints, we estimated that long term semen storage requires 10% dimethyl sulfoxide as a protective agent, −1 °C/min chilling rate (18 °C to 5 °C) and −13 °C/min freezing rate (5 °C to −80 °C), followed by immersion in liquid nitrogen.
Keywords:Ciona intestinalis  Spermatozoon  Cryopreservation  Dimethyl sulfoxide  Fertilization success
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