Magnetic separation of peripheral nerve-resident cells underscores key molecular features of human Schwann cells and fibroblasts: an immunochemical and transcriptomics approach

Autores
Peng, Kaiwen; Sant, David; Andersen, Natalia Denise; Silvera, Risset; Camarena, Vladimir; Piñero, Gonzalo Miguel; Graham, Regina; Khan, Aisha; Xu, Xiao Ming; Wang, Gaofeng; Monje, Paula
Año de publicación
2020
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Nerve-derived human Schwann cell (SC) cultures are irreplaceable models for basic and translational research but their use can be limited due to the risk of fibroblast overgrowth. Fibroblasts are an ill-defined population consisting of highly proliferative cells that, contrary to human SCs, do not undergo senescence in culture. We initiated this study by performing an exhaustive immunological and functional characterization of adult nerve-derived human SCs and fibroblasts to reveal their properties and optimize a protocol of magnetic-activated cell sorting (MACS) to separate them effectively both as viable and biologically competent cells. We next used immunofluorescence microscopy imaging, flow cytometry analysis and next generation RNA sequencing (RNA-seq) to unambiguously characterize the post-MACS cell products. High resolution transcriptome profiling revealed the identity of key lineage-specific transcripts and the clearly distinct neural crest and mesenchymal origin of human SCs and fibroblasts, respectively. Our analysis underscored a progenitor- or stem cell-like molecular phenotype in SCs and fibroblasts and the heterogeneity of the fibroblast populations. In addition, pathway analysis of RNA-seq data highlighted putative bidirectional networks of fibroblast-to-SC signaling that predict a complementary, yet seemingly independent contribution of SCs and fibroblasts to nerve regeneration. In sum, combining MACS with immunochemical and transcriptomics approaches provides an ideal workflow to exhaustively assess the identity, the stage of differentiation and functional features of highly purified cells from human peripheral nerve tissues.
Fil: Peng, Kaiwen. Indiana University. School of Medicine; Estados Unidos. Nanfang Hospital; China
Fil: Sant, David. University of Utah; Estados Unidos. Miami University. School of Medicine; Estados Unidos
Fil: Andersen, Natalia Denise. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina. Miami University. School of Medicine; Estados Unidos
Fil: Silvera, Risset. Miami University. School of Medicine; Estados Unidos
Fil: Camarena, Vladimir. Miami University. School of Medicine; Estados Unidos
Fil: Piñero, Gonzalo Miguel. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas "Prof. Alejandro C. Paladini". Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Físico-Química Biológicas; Argentina. Miami University. School of Medicine; Estados Unidos
Fil: Graham, Regina. Miami University. School of Medicine; Estados Unidos
Fil: Khan, Aisha. Miami University. School of Medicine; Estados Unidos
Fil: Xu, Xiao Ming. Indiana University. School of Medicine; Estados Unidos
Fil: Wang, Gaofeng. Miami University. School of Medicine; Estados Unidos
Fil: Monje, Paula. Indiana University. School of Medicine; Estados Unidos. Miami University. School of Medicine; Estados Unidos
Materia
HUMAN SCHWANN CELLS
GLIAL BIOLOGY
GENOMIC ANALYSIS
NEUROSCIENCE
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/120021
Acceder
id CONICETDig_72409fe33d8212ff4c8015715e5ae80a
oai_identifier_str oai:ri.conicet.gov.ar:11336/120021
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Magnetic separation of peripheral nerve-resident cells underscores key molecular features of human Schwann cells and fibroblasts: an immunochemical and transcriptomics approachPeng, KaiwenSant, DavidAndersen, Natalia DeniseSilvera, RissetCamarena, VladimirPiñero, Gonzalo MiguelGraham, ReginaKhan, AishaXu, Xiao MingWang, GaofengMonje, PaulaHUMAN SCHWANN CELLSGLIAL BIOLOGYGENOMIC ANALYSISNEUROSCIENCEhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Nerve-derived human Schwann cell (SC) cultures are irreplaceable models for basic and translational research but their use can be limited due to the risk of fibroblast overgrowth. Fibroblasts are an ill-defined population consisting of highly proliferative cells that, contrary to human SCs, do not undergo senescence in culture. We initiated this study by performing an exhaustive immunological and functional characterization of adult nerve-derived human SCs and fibroblasts to reveal their properties and optimize a protocol of magnetic-activated cell sorting (MACS) to separate them effectively both as viable and biologically competent cells. We next used immunofluorescence microscopy imaging, flow cytometry analysis and next generation RNA sequencing (RNA-seq) to unambiguously characterize the post-MACS cell products. High resolution transcriptome profiling revealed the identity of key lineage-specific transcripts and the clearly distinct neural crest and mesenchymal origin of human SCs and fibroblasts, respectively. Our analysis underscored a progenitor- or stem cell-like molecular phenotype in SCs and fibroblasts and the heterogeneity of the fibroblast populations. In addition, pathway analysis of RNA-seq data highlighted putative bidirectional networks of fibroblast-to-SC signaling that predict a complementary, yet seemingly independent contribution of SCs and fibroblasts to nerve regeneration. In sum, combining MACS with immunochemical and transcriptomics approaches provides an ideal workflow to exhaustively assess the identity, the stage of differentiation and functional features of highly purified cells from human peripheral nerve tissues.Fil: Peng, Kaiwen. Indiana University. School of Medicine; Estados Unidos. Nanfang Hospital; ChinaFil: Sant, David. University of Utah; Estados Unidos. Miami University. School of Medicine; Estados UnidosFil: Andersen, Natalia Denise. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina. Miami University. School of Medicine; Estados UnidosFil: Silvera, Risset. Miami University. School of Medicine; Estados UnidosFil: Camarena, Vladimir. Miami University. School of Medicine; Estados UnidosFil: Piñero, Gonzalo Miguel. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas "Prof. Alejandro C. Paladini". Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Físico-Química Biológicas; Argentina. Miami University. School of Medicine; Estados UnidosFil: Graham, Regina. Miami University. School of Medicine; Estados UnidosFil: Khan, Aisha. Miami University. School of Medicine; Estados UnidosFil: Xu, Xiao Ming. Indiana University. School of Medicine; Estados UnidosFil: Wang, Gaofeng. Miami University. School of Medicine; Estados UnidosFil: Monje, Paula. Indiana University. School of Medicine; Estados Unidos. Miami University. School of Medicine; Estados UnidosNature Publishing Group2020-12-28info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/120021Peng, Kaiwen; Sant, David; Andersen, Natalia Denise; Silvera, Risset; Camarena, Vladimir; et al.; Magnetic separation of peripheral nerve-resident cells underscores key molecular features of human Schwann cells and fibroblasts: an immunochemical and transcriptomics approach; Nature Publishing Group; Scientific Reports; 10; 1; 28-12-2020; 1-202045-2322CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://www.nature.com/articles/s41598-020-74128-3info:eu-repo/semantics/altIdentifier/doi/10.1038/s41598-020-74128-3info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-03T09:52:26Zoai:ri.conicet.gov.ar:11336/120021instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-09-03 09:52:26.636CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Magnetic separation of peripheral nerve-resident cells underscores key molecular features of human Schwann cells and fibroblasts: an immunochemical and transcriptomics approach
title Magnetic separation of peripheral nerve-resident cells underscores key molecular features of human Schwann cells and fibroblasts: an immunochemical and transcriptomics approach
spellingShingle Magnetic separation of peripheral nerve-resident cells underscores key molecular features of human Schwann cells and fibroblasts: an immunochemical and transcriptomics approach
Peng, Kaiwen
HUMAN SCHWANN CELLS
GLIAL BIOLOGY
GENOMIC ANALYSIS
NEUROSCIENCE
title_short Magnetic separation of peripheral nerve-resident cells underscores key molecular features of human Schwann cells and fibroblasts: an immunochemical and transcriptomics approach
title_full Magnetic separation of peripheral nerve-resident cells underscores key molecular features of human Schwann cells and fibroblasts: an immunochemical and transcriptomics approach
title_fullStr Magnetic separation of peripheral nerve-resident cells underscores key molecular features of human Schwann cells and fibroblasts: an immunochemical and transcriptomics approach
title_full_unstemmed Magnetic separation of peripheral nerve-resident cells underscores key molecular features of human Schwann cells and fibroblasts: an immunochemical and transcriptomics approach
title_sort Magnetic separation of peripheral nerve-resident cells underscores key molecular features of human Schwann cells and fibroblasts: an immunochemical and transcriptomics approach
dc.creator.none.fl_str_mv Peng, Kaiwen
Sant, David
Andersen, Natalia Denise
Silvera, Risset
Camarena, Vladimir
Piñero, Gonzalo Miguel
Graham, Regina
Khan, Aisha
Xu, Xiao Ming
Wang, Gaofeng
Monje, Paula
author Peng, Kaiwen
author_facet Peng, Kaiwen
Sant, David
Andersen, Natalia Denise
Silvera, Risset
Camarena, Vladimir
Piñero, Gonzalo Miguel
Graham, Regina
Khan, Aisha
Xu, Xiao Ming
Wang, Gaofeng
Monje, Paula
author_role author
author2 Sant, David
Andersen, Natalia Denise
Silvera, Risset
Camarena, Vladimir
Piñero, Gonzalo Miguel
Graham, Regina
Khan, Aisha
Xu, Xiao Ming
Wang, Gaofeng
Monje, Paula
author2_role author
author
author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv HUMAN SCHWANN CELLS
GLIAL BIOLOGY
GENOMIC ANALYSIS
NEUROSCIENCE
topic HUMAN SCHWANN CELLS
GLIAL BIOLOGY
GENOMIC ANALYSIS
NEUROSCIENCE
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Nerve-derived human Schwann cell (SC) cultures are irreplaceable models for basic and translational research but their use can be limited due to the risk of fibroblast overgrowth. Fibroblasts are an ill-defined population consisting of highly proliferative cells that, contrary to human SCs, do not undergo senescence in culture. We initiated this study by performing an exhaustive immunological and functional characterization of adult nerve-derived human SCs and fibroblasts to reveal their properties and optimize a protocol of magnetic-activated cell sorting (MACS) to separate them effectively both as viable and biologically competent cells. We next used immunofluorescence microscopy imaging, flow cytometry analysis and next generation RNA sequencing (RNA-seq) to unambiguously characterize the post-MACS cell products. High resolution transcriptome profiling revealed the identity of key lineage-specific transcripts and the clearly distinct neural crest and mesenchymal origin of human SCs and fibroblasts, respectively. Our analysis underscored a progenitor- or stem cell-like molecular phenotype in SCs and fibroblasts and the heterogeneity of the fibroblast populations. In addition, pathway analysis of RNA-seq data highlighted putative bidirectional networks of fibroblast-to-SC signaling that predict a complementary, yet seemingly independent contribution of SCs and fibroblasts to nerve regeneration. In sum, combining MACS with immunochemical and transcriptomics approaches provides an ideal workflow to exhaustively assess the identity, the stage of differentiation and functional features of highly purified cells from human peripheral nerve tissues.
Fil: Peng, Kaiwen. Indiana University. School of Medicine; Estados Unidos. Nanfang Hospital; China
Fil: Sant, David. University of Utah; Estados Unidos. Miami University. School of Medicine; Estados Unidos
Fil: Andersen, Natalia Denise. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina. Miami University. School of Medicine; Estados Unidos
Fil: Silvera, Risset. Miami University. School of Medicine; Estados Unidos
Fil: Camarena, Vladimir. Miami University. School of Medicine; Estados Unidos
Fil: Piñero, Gonzalo Miguel. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas "Prof. Alejandro C. Paladini". Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Físico-Química Biológicas; Argentina. Miami University. School of Medicine; Estados Unidos
Fil: Graham, Regina. Miami University. School of Medicine; Estados Unidos
Fil: Khan, Aisha. Miami University. School of Medicine; Estados Unidos
Fil: Xu, Xiao Ming. Indiana University. School of Medicine; Estados Unidos
Fil: Wang, Gaofeng. Miami University. School of Medicine; Estados Unidos
Fil: Monje, Paula. Indiana University. School of Medicine; Estados Unidos. Miami University. School of Medicine; Estados Unidos
description Nerve-derived human Schwann cell (SC) cultures are irreplaceable models for basic and translational research but their use can be limited due to the risk of fibroblast overgrowth. Fibroblasts are an ill-defined population consisting of highly proliferative cells that, contrary to human SCs, do not undergo senescence in culture. We initiated this study by performing an exhaustive immunological and functional characterization of adult nerve-derived human SCs and fibroblasts to reveal their properties and optimize a protocol of magnetic-activated cell sorting (MACS) to separate them effectively both as viable and biologically competent cells. We next used immunofluorescence microscopy imaging, flow cytometry analysis and next generation RNA sequencing (RNA-seq) to unambiguously characterize the post-MACS cell products. High resolution transcriptome profiling revealed the identity of key lineage-specific transcripts and the clearly distinct neural crest and mesenchymal origin of human SCs and fibroblasts, respectively. Our analysis underscored a progenitor- or stem cell-like molecular phenotype in SCs and fibroblasts and the heterogeneity of the fibroblast populations. In addition, pathway analysis of RNA-seq data highlighted putative bidirectional networks of fibroblast-to-SC signaling that predict a complementary, yet seemingly independent contribution of SCs and fibroblasts to nerve regeneration. In sum, combining MACS with immunochemical and transcriptomics approaches provides an ideal workflow to exhaustively assess the identity, the stage of differentiation and functional features of highly purified cells from human peripheral nerve tissues.
publishDate 2020
dc.date.none.fl_str_mv 2020-12-28
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/120021
Peng, Kaiwen; Sant, David; Andersen, Natalia Denise; Silvera, Risset; Camarena, Vladimir; et al.; Magnetic separation of peripheral nerve-resident cells underscores key molecular features of human Schwann cells and fibroblasts: an immunochemical and transcriptomics approach; Nature Publishing Group; Scientific Reports; 10; 1; 28-12-2020; 1-20
2045-2322
CONICET Digital
CONICET
url http://hdl.handle.net/11336/120021
identifier_str_mv Peng, Kaiwen; Sant, David; Andersen, Natalia Denise; Silvera, Risset; Camarena, Vladimir; et al.; Magnetic separation of peripheral nerve-resident cells underscores key molecular features of human Schwann cells and fibroblasts: an immunochemical and transcriptomics approach; Nature Publishing Group; Scientific Reports; 10; 1; 28-12-2020; 1-20
2045-2322
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/https://www.nature.com/articles/s41598-020-74128-3
info:eu-repo/semantics/altIdentifier/doi/10.1038/s41598-020-74128-3
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
dc.publisher.none.fl_str_mv Nature Publishing Group
publisher.none.fl_str_mv Nature Publishing Group
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
_version_ 1842269158219186176
score 13.13397

Publicaciones similares