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

Autor: Xiao Ming Xu, Risset Silvera, Vladimir Camarena, Gaofeng Wang, Kaiwen Peng, Regina M. Graham, Natalia Denise Andersen, David W. Sant, Gonzalo Piñero, Aisha Khan, Paula V. Monje
Jazyk: angličtina
Rok vydání: 2020
Předmět:
Male
Cell Culture Techniques
lcsh:Medicine
Cell Separation
Workflow
Genomic analysis
Immunological techniques
purl.org/becyt/ford/1 [https]
Transcriptome
Cluster Analysis
Child
lcsh:Science
education.field_of_study
Multidisciplinary
Biological techniques
Neural crest
Bioquímica y Biología Molecular
Middle Aged
Cell sorting
Cell biology
medicine.anatomical_structure
NEUROSCIENCE
Isolation
separation and purification
Female
CIENCIAS NATURALES Y EXACTAS
Cell signalling
Adult
Adolescent
Primary Cell Culture
Population
Schwann cell
Biology
Molecular neuroscience
Senescence
GLIAL BIOLOGY
Article
Cell Line
Ciencias Biológicas
Young Adult
medicine
Humans
Peripheral Nerves
Regeneration and repair in the nervous system
purl.org/becyt/ford/1.6 [https]
education
Fibroblast
Aged
Biological models
Gene Expression Profiling
Regeneration (biology)
Mesenchymal stem cell
lcsh:R
Glial biology
HUMAN SCHWANN CELLS
Fibroblasts
Cellular neuroscience
GENOMIC ANALYSIS
Nerve Regeneration
lcsh:Q
Schwann Cells
Peripheral nervous system
Neuroscience
Zdroj: Scientific Reports, Vol 10, Iss 1, Pp 1-20 (2020)
CONICET Digital (CONICET)
Consejo Nacional de Investigaciones Científicas y Técnicas
instacron:CONICET
Scientific Reports
ISSN: 2045-2322
Popis: 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
Databáze: OpenAIRE
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