Effect of spermatozoa motility hyperactivation factors and gamete coincubation duration on in vitro bovine embryo development using flow cytometrically sorted spermatozoa
Autor: | Pablo J. Ross, Cristóbal Fresno, Hugo Hector Ortega, M. E. Kjelland, James L. Chitwood, Luis B. Ferré, Y. S. Bogliotti |
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Jazyk: | angličtina |
Rok vydání: | 2017 |
Předmět: |
Male
medicine.medical_treatment Embryonic Development Reproductive technology Cell Separation Fertilization in Vitro Biology Andrology Embryo Culture Techniques Ciencias Biológicas 03 medical and health sciences 0302 clinical medicine Endocrinology Caffeine Genetics medicine Animals Blastocyst Molecular Biology Sperm motility Fertilisation 030219 obstetrics & reproductive medicine In vitro fertilisation 0402 animal and dairy science Embryo culture 04 agricultural and veterinary sciences Anatomy SEXED SPERMATOZOA Polyspermy Flow Cytometry 040201 dairy & animal science Spermatozoa medicine.anatomical_structure Reproductive Medicine IVF Sperm Motility Animal Science and Zoology Cattle Female Biología Reproductiva Embryo quality CIENCIAS NATURALES Y EXACTAS Developmental Biology Biotechnology |
Popis: | The aim of the present study was to evaluate the effects of sperm motility enhancers and different IVF times on cleavage, polyspermy, blastocyst formation, embryo quality and hatching ability. In Experiment 1, sex-sorted X chromosome-bearing Bos taurus spermatozoa were incubated for 30 min before 18 h fertilisation with hyperactivating factors, namely 10mM caffeine (CA), 5mMtheophylline (TH), 10mM caffeine and 5mMtheophylline (CA+TH); and untreated spermatozoa (control). In Experiment 2, matured B.Taurus oocytes were fertilised using a short (8 h) or standard (18 h) fertilisation length, comparing two different fertilisation media, namely synthetic oviducal fluid (SOF) fertilisation medium (SOF-FERT) and M199 fertilisation medium (M199-FERT). Cleavage and blastocyst formation rates were significantly higher in the CA+TH group (77% and 27%, respectively) compared with the control group (71% and 21%, respectively). Cleavage rates and blastocyst formation were significantly lower for the shortest fertilisation time (8 h) in M199-FERT medium (42% and 12%, respectively). The SOF-FERT medium with an 8 h fertilisation time resulted in the highest cleavage rates and blastocyst formation (74% and 29%, respectively). The SOF-FERT medium produced the highest embryo quality (50% Grade 1) and hatching rate (66%). Motility enhancers did not affect polyspermy rates, whereas polyspermy was affected when fertilisation length was extended from 8 h (3%) to 18 h (9%) and in M199-FERT (14%) compared with SOF-FERT (6%). We conclude that adding the motility enhancers CA and TH to sex sorted spermatozoa and Tyrode´s albumin lactate pyruvate (TALP)-Sperm can improve cleavage and embryo development rates without increasing polyspermy. In addition, shortening the oocyte-sperm coincubation time (8 h) resulted in similar overall embryo performance rates compared with the prolonged (18 h) interval. Fil: Ferré, Luis Bernardo. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Buenos Aires; Argentina Fil: Bogliotti, Yanina. University of California; Estados Unidos Fil: Chitwood, James L.. University of California; Estados Unidos Fil: Fresno Rodríguez, Cristóbal. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Católica de Córdoba; Argentina Fil: Ortega, Hugo Hector. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina Fil: Kjelland, Michael E.. Conservation, Genetics and Biotech; Estados Unidos Fil: Ross, Pablo J.. University of California; Estados Unidos |
Databáze: | OpenAIRE |
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