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Comparative study of sperm washing and selection methods after cryopreservation and its influence on sperm subpopulational structure in a bovine model.

The effect of different sperm washing-selection methods on sperm morphometric characteristics as a study to detect differences in the subpopulational structure has been carried out in detail in a bovine model. Cryopreserved sperm samples from 5 bulls were thawed, pooled, and processed by TALP-washin...

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Autor Principal:	Garc?a Herreros, Manuel
Formato:	Artículos
Lenguaje:	eng
Publicado:	2016
Materias:	BOVINE MODEL PRINCIPAL COMPONENT ANALYSIS SPERM SELECTION SPERM SUBPOPULATIONAL
Acceso en línea:	http://repositorio.educacionsuperior.gob.ec/handle/28000/2714
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spelling	oai:localhost:28000-27142017-04-06T15:03:04Z Comparative study of sperm washing and selection methods after cryopreservation and its influence on sperm subpopulational structure in a bovine model. Garc?a Herreros, Manuel BOVINE MODEL PRINCIPAL COMPONENT ANALYSIS SPERM SELECTION SPERM SUBPOPULATIONAL The effect of different sperm washing-selection methods on sperm morphometric characteristics as a study to detect differences in the subpopulational structure has been carried out in detail in a bovine model. Cryopreserved sperm samples from 5 bulls were thawed, pooled, and processed by TALP-washing centrifugation method (TWCM), selective Percoll discontinuous density-gradient centrifugation method (PDGM), and self-migration swim-up separation method (SUMM). Live-dead assay (SYBR-14/ethidium homodimer-1), chlortetracycline assay (CTC), and sperm motility were assessed, and aliquots of sperm were processed for automated sperm morphometry analysis (ASMA) simultaneously before (raw thawed sperm used as control, RTS) and after different sperm washing-selection techniques. Deleterious effects of different methods were evident, particularly on sperm membrane integrity (p?<?0.05) and capacitation status (p?<?0.05). Moreover, each cell was measured for four primary dimensional parameters, and three shape parameters. All sperm morphometric parameters evaluated were analyzed by principal component analysis (PCA) and multivariate clustering analyses. PCA revealed two principal components for each sperm washing or separation method explaining more than the 91% of the variance. The number of subpopulations found was the same for all methods (four) except for PDGM (three). However, irrespective of the number of subpopulations defined by PCA and clustering analyses, the sperm subpopulational structure was found to be different and strongly influenced by the sperm selection procedure due to statistical differences found regarding the sperm biophysical changes induced by each method used (p?<?0.001). It is concluded that different sperm washing-selection methods commonly used during IVF process, may lead to alterations in sperm morphometric characteristics, which might explain the different results seen after IVF, since an important influence of these methods on sperm subpopulational structure has been demonstrated. Universidad T?cnica Del Cotopaxi http://www.ncbi.nlm.nih.gov/pubmed/25007138 2016-11-08T17:37:46Z 2016-11-08T17:37:46Z 2014 article Garc?a Herreros, Manuel. Leal CL. (2014). Comparative study of sperm washing and selection methods after cryopreservation and its influence on sperm subpopulational structure in a bovine model. Systems Biology in Reproductive Medicine Volumen 60. pp. 338-347 1939-6368 http://repositorio.educacionsuperior.gob.ec/handle/28000/2714 eng openAccess pp. 338-347
institution	SENESCYT
collection	Repositorio SENESCYT
biblioteca	Biblioteca Senescyt
language	eng
format	Artículos
topic	BOVINE MODEL PRINCIPAL COMPONENT ANALYSIS SPERM SELECTION SPERM SUBPOPULATIONAL
spellingShingle	BOVINE MODEL PRINCIPAL COMPONENT ANALYSIS SPERM SELECTION SPERM SUBPOPULATIONAL Garc?a Herreros, Manuel Comparative study of sperm washing and selection methods after cryopreservation and its influence on sperm subpopulational structure in a bovine model.
description	The effect of different sperm washing-selection methods on sperm morphometric characteristics as a study to detect differences in the subpopulational structure has been carried out in detail in a bovine model. Cryopreserved sperm samples from 5 bulls were thawed, pooled, and processed by TALP-washing centrifugation method (TWCM), selective Percoll discontinuous density-gradient centrifugation method (PDGM), and self-migration swim-up separation method (SUMM). Live-dead assay (SYBR-14/ethidium homodimer-1), chlortetracycline assay (CTC), and sperm motility were assessed, and aliquots of sperm were processed for automated sperm morphometry analysis (ASMA) simultaneously before (raw thawed sperm used as control, RTS) and after different sperm washing-selection techniques. Deleterious effects of different methods were evident, particularly on sperm membrane integrity (p?<?0.05) and capacitation status (p?<?0.05). Moreover, each cell was measured for four primary dimensional parameters, and three shape parameters. All sperm morphometric parameters evaluated were analyzed by principal component analysis (PCA) and multivariate clustering analyses. PCA revealed two principal components for each sperm washing or separation method explaining more than the 91% of the variance. The number of subpopulations found was the same for all methods (four) except for PDGM (three). However, irrespective of the number of subpopulations defined by PCA and clustering analyses, the sperm subpopulational structure was found to be different and strongly influenced by the sperm selection procedure due to statistical differences found regarding the sperm biophysical changes induced by each method used (p?<?0.001). It is concluded that different sperm washing-selection methods commonly used during IVF process, may lead to alterations in sperm morphometric characteristics, which might explain the different results seen after IVF, since an important influence of these methods on sperm subpopulational structure has been demonstrated.
author	Garc?a Herreros, Manuel
author_facet	Garc?a Herreros, Manuel
author_sort	Garc?a Herreros, Manuel
title	Comparative study of sperm washing and selection methods after cryopreservation and its influence on sperm subpopulational structure in a bovine model.
title_short	Comparative study of sperm washing and selection methods after cryopreservation and its influence on sperm subpopulational structure in a bovine model.
title_full	Comparative study of sperm washing and selection methods after cryopreservation and its influence on sperm subpopulational structure in a bovine model.
title_fullStr	Comparative study of sperm washing and selection methods after cryopreservation and its influence on sperm subpopulational structure in a bovine model.
title_full_unstemmed	Comparative study of sperm washing and selection methods after cryopreservation and its influence on sperm subpopulational structure in a bovine model.
title_sort	comparative study of sperm washing and selection methods after cryopreservation and its influence on sperm subpopulational structure in a bovine model.
publishDate	2016
url	http://repositorio.educacionsuperior.gob.ec/handle/28000/2714
_version_	1634995079164723200
score	11,871979

Comparative study of sperm washing and selection methods after cryopreservation and its influence on sperm subpopulational structure in a bovine model.

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