Multiple flow cytometry analysis for assessing human sperm functional characteristics
Autor(a) principal: | |
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Data de Publicação: | 2023 |
Outros Autores: | , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Institucional da UNESP |
Texto Completo: | http://dx.doi.org/10.1016/j.reprotox.2023.108353 http://hdl.handle.net/11449/248384 |
Resumo: | Multiparametric analysis by flow cytometry solves one of the major problems in sperm evaluation, the inability to test multiple attributes simultaneously in a single cell, which would increase the precision to predict fertility potential since several sperm parameters are tested. The association of fluorochromes and compounds conjugated to fluorochromes in multiparametric sperm analysis is well-established in microscopy techniques. However, these techniques are subjective and limit the assessment in small cell numbers, thereby harming analytic accuracy. Therefore, the current study aimed to present new possibilities for assessing the integrity and stability of the sperm plasma membrane, acrosome status, mitochondrial potential, and superoxide anion production in the mitochondrial matrix in only 2 cytometric assays using cytometers equipped with 2 and 3 lasers. For this, human semen samples collected by masturbation and selected by the swim-up technique were divided into 3 treatments: T0 (flash-frozen semen), T50 (flash-frozen semen + fresh semen, V: V), and T100 (fresh semen) for the validation of the multiparametric protocols by flow cytometry. For both protocols, sperm percentage with positive stain for all fluorophores differed significantly between treatments. The determination coefficients presented values close to 1, which validated objective, sensitive, rapid, and reproducible methodologies. Therefore, we concluded that the results reflect the status of analyzed structure, enabling a more accurate diagnosis of male infertility that has become an increasingly prevalent worldwide setback due to exposure to a variety of environmental toxicants. |
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Multiple flow cytometry analysis for assessing human sperm functional characteristicsFlow cytometryHuman spermMale infertilityMultiparametric analysisReproductive toxicityMultiparametric analysis by flow cytometry solves one of the major problems in sperm evaluation, the inability to test multiple attributes simultaneously in a single cell, which would increase the precision to predict fertility potential since several sperm parameters are tested. The association of fluorochromes and compounds conjugated to fluorochromes in multiparametric sperm analysis is well-established in microscopy techniques. However, these techniques are subjective and limit the assessment in small cell numbers, thereby harming analytic accuracy. Therefore, the current study aimed to present new possibilities for assessing the integrity and stability of the sperm plasma membrane, acrosome status, mitochondrial potential, and superoxide anion production in the mitochondrial matrix in only 2 cytometric assays using cytometers equipped with 2 and 3 lasers. For this, human semen samples collected by masturbation and selected by the swim-up technique were divided into 3 treatments: T0 (flash-frozen semen), T50 (flash-frozen semen + fresh semen, V: V), and T100 (fresh semen) for the validation of the multiparametric protocols by flow cytometry. For both protocols, sperm percentage with positive stain for all fluorophores differed significantly between treatments. The determination coefficients presented values close to 1, which validated objective, sensitive, rapid, and reproducible methodologies. Therefore, we concluded that the results reflect the status of analyzed structure, enabling a more accurate diagnosis of male infertility that has become an increasingly prevalent worldwide setback due to exposure to a variety of environmental toxicants.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Institute of Biosciences Department of Structural and Functional Biology Laboratory of Reproductive and Developmental Biology and Toxicology São Paulo State University (UNESP), São PauloSchool of Veterinary Medicine and Animal Science Department of Veterinary Surgery and Animal Reproduction São Paulo State University (UNESP), São PauloInstitute of Biosciences Department of Structural and Functional Biology Laboratory of Reproductive and Developmental Biology and Toxicology São Paulo State University (UNESP), São PauloSchool of Veterinary Medicine and Animal Science Department of Veterinary Surgery and Animal Reproduction São Paulo State University (UNESP), São PauloFAPESP: 2015/13941–3Universidade Estadual Paulista (UNESP)de Lima Rosa, Josiane [UNESP]de Paula Freitas Dell'Aqua, Camila [UNESP]de Souza, Fabiana Ferreira [UNESP]Missassi, Gabriela [UNESP]Kempinas, Wilma De Grava [UNESP]2023-07-29T13:42:35Z2023-07-29T13:42:35Z2023-04-01info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articlehttp://dx.doi.org/10.1016/j.reprotox.2023.108353Reproductive Toxicology, v. 117.1873-17080890-6238http://hdl.handle.net/11449/24838410.1016/j.reprotox.2023.1083532-s2.0-85148371459Scopusreponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPengReproductive Toxicologyinfo:eu-repo/semantics/openAccess2023-07-29T13:42:36Zoai:repositorio.unesp.br:11449/248384Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462023-07-29T13:42:36Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false |
dc.title.none.fl_str_mv |
Multiple flow cytometry analysis for assessing human sperm functional characteristics |
title |
Multiple flow cytometry analysis for assessing human sperm functional characteristics |
spellingShingle |
Multiple flow cytometry analysis for assessing human sperm functional characteristics de Lima Rosa, Josiane [UNESP] Flow cytometry Human sperm Male infertility Multiparametric analysis Reproductive toxicity |
title_short |
Multiple flow cytometry analysis for assessing human sperm functional characteristics |
title_full |
Multiple flow cytometry analysis for assessing human sperm functional characteristics |
title_fullStr |
Multiple flow cytometry analysis for assessing human sperm functional characteristics |
title_full_unstemmed |
Multiple flow cytometry analysis for assessing human sperm functional characteristics |
title_sort |
Multiple flow cytometry analysis for assessing human sperm functional characteristics |
author |
de Lima Rosa, Josiane [UNESP] |
author_facet |
de Lima Rosa, Josiane [UNESP] de Paula Freitas Dell'Aqua, Camila [UNESP] de Souza, Fabiana Ferreira [UNESP] Missassi, Gabriela [UNESP] Kempinas, Wilma De Grava [UNESP] |
author_role |
author |
author2 |
de Paula Freitas Dell'Aqua, Camila [UNESP] de Souza, Fabiana Ferreira [UNESP] Missassi, Gabriela [UNESP] Kempinas, Wilma De Grava [UNESP] |
author2_role |
author author author author |
dc.contributor.none.fl_str_mv |
Universidade Estadual Paulista (UNESP) |
dc.contributor.author.fl_str_mv |
de Lima Rosa, Josiane [UNESP] de Paula Freitas Dell'Aqua, Camila [UNESP] de Souza, Fabiana Ferreira [UNESP] Missassi, Gabriela [UNESP] Kempinas, Wilma De Grava [UNESP] |
dc.subject.por.fl_str_mv |
Flow cytometry Human sperm Male infertility Multiparametric analysis Reproductive toxicity |
topic |
Flow cytometry Human sperm Male infertility Multiparametric analysis Reproductive toxicity |
description |
Multiparametric analysis by flow cytometry solves one of the major problems in sperm evaluation, the inability to test multiple attributes simultaneously in a single cell, which would increase the precision to predict fertility potential since several sperm parameters are tested. The association of fluorochromes and compounds conjugated to fluorochromes in multiparametric sperm analysis is well-established in microscopy techniques. However, these techniques are subjective and limit the assessment in small cell numbers, thereby harming analytic accuracy. Therefore, the current study aimed to present new possibilities for assessing the integrity and stability of the sperm plasma membrane, acrosome status, mitochondrial potential, and superoxide anion production in the mitochondrial matrix in only 2 cytometric assays using cytometers equipped with 2 and 3 lasers. For this, human semen samples collected by masturbation and selected by the swim-up technique were divided into 3 treatments: T0 (flash-frozen semen), T50 (flash-frozen semen + fresh semen, V: V), and T100 (fresh semen) for the validation of the multiparametric protocols by flow cytometry. For both protocols, sperm percentage with positive stain for all fluorophores differed significantly between treatments. The determination coefficients presented values close to 1, which validated objective, sensitive, rapid, and reproducible methodologies. Therefore, we concluded that the results reflect the status of analyzed structure, enabling a more accurate diagnosis of male infertility that has become an increasingly prevalent worldwide setback due to exposure to a variety of environmental toxicants. |
publishDate |
2023 |
dc.date.none.fl_str_mv |
2023-07-29T13:42:35Z 2023-07-29T13:42:35Z 2023-04-01 |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://dx.doi.org/10.1016/j.reprotox.2023.108353 Reproductive Toxicology, v. 117. 1873-1708 0890-6238 http://hdl.handle.net/11449/248384 10.1016/j.reprotox.2023.108353 2-s2.0-85148371459 |
url |
http://dx.doi.org/10.1016/j.reprotox.2023.108353 http://hdl.handle.net/11449/248384 |
identifier_str_mv |
Reproductive Toxicology, v. 117. 1873-1708 0890-6238 10.1016/j.reprotox.2023.108353 2-s2.0-85148371459 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
Reproductive Toxicology |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.source.none.fl_str_mv |
Scopus reponame:Repositório Institucional da UNESP instname:Universidade Estadual Paulista (UNESP) instacron:UNESP |
instname_str |
Universidade Estadual Paulista (UNESP) |
instacron_str |
UNESP |
institution |
UNESP |
reponame_str |
Repositório Institucional da UNESP |
collection |
Repositório Institucional da UNESP |
repository.name.fl_str_mv |
Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP) |
repository.mail.fl_str_mv |
|
_version_ |
1799965319941324800 |