Selection of non-apoptotic sperm by magnetic activated cell sorting in Senegalese sole (Solea senegalensis)
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Autor/esValcarce, D.G. (David G); Herráez, P.; Chereguini-Fernández-Maquieira, M.O. (María del Olvido); Rodríguez, C.; Robles, V. (Vanesa)
Senegalese sole (Solea senegalensis) is a promising species in aquaculture. However, due to decreased sperm quality in F1 generations and the absence of courtship in those individuals born in captivity, artificial fertilization is being used to generate new progenies. The objective of this study was to implement a sperm selection method for non-apoptotic sperm subpopulation recovery prior to sperm cryopreservation. In particular, magnetic activated cell sorting (MACS) is used to eliminate apoptotic spermatozoa. This study represents the proof-of-concept for MACS applicability in teleost species relevant in aquaculture. Apoptotic cell population was studied by flow cytometry using YO-PRO 1 and a caspase detection kit. Also, reactive oxygen species were measured in sperm samples. Our data demonstrated that caspase detection is more specific than YO-PRO 1 in the identification of apoptotic cells in S. senegalensis seminal samples. The results showed that the percentage of apoptotic cells ...