Human Schwann cell senescence is not prevented by ectopic expression of human telomerase reverse transcriptase
- Autores
- Andersen, Natalia Denise; Kuo, Blanche; Piñero, Gonzalo Miguel; Ravelo, Kristine; Rai, Priyamvada; Monje, Paula
- Año de publicación
- 2016
- Idioma
- inglés
- Tipo de recurso
- documento de conferencia
- Estado
- versión publicada
- Descripción
- Isolated human Schwann cells (hSCs) typically become senescent and unresponsive to mitogenic factors with continued in vitro expansion. Data from our lab has shown that adult nerve-derived hSCs, in contrast to rodent (rat) SCs, inevitably stop proliferating and acquire a senescent phenotype characterized by high levels of senescence-associated (SA)-β galactosidase activity and morphological changes that include cell enlargement and appearance of multinucleated cells. As opposed to rodent SCs, hSC cultures consist of mixed populations of proliferating cells, senescent cells and cells at different stages of differentiation regardless of the nerve of origin and other donor-specific factors. RNA-seq analysis of representative cultures of hSCs did not reveal the presence of telomerase reverse transcriptase (TERT) mRNA while other TERT-related genes (e.g. TERF1, telomeric repeat binding factor, and TEP1, telomeraseassociated protein) were well-represented in the hSC transcriptome. In an attempt to overcome senescence, we used retroviral vectors and antibiotic selection to generate hSC lines ectopically expressing human (h)-TERT. For these experiments, highly proliferative, non-senescent, early passage hSC cultures were stably transduced with the retroviruses h-TERT-hygro or h-TERTpuro, each encoding the h-TERT gene along with hygromycin or puromycin resistance genes, respectively. Transduced hSC cultures from three different donors were selected and subjected to three rounds of expansion in medium containing chemical mitogens. Subsequently, the cultures were analyzed for their rate of proliferation by means of EdU incorporation assays and the acquisition of senescence by means of SA-β galactosidase activity assays in each round. We found that whereas ectopic h-TERT expression extended the lifespan of cultured hSCs when compared to non-infected or GFP-expressing cells, it was not sufficient to confer immortalization and overcome senescence. In sum, our results suggest that progression of the hSCs to a senescent state likely is stress-induced rather than dependent on replication-associated telomere shortening
Fil: Andersen, Natalia Denise. Miami University; Estados Unidos. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Kuo, Blanche. Miami University; Estados Unidos
Fil: Piñero, Gonzalo Miguel. Miami University; Estados Unidos. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Ravelo, Kristine. Miami University; Estados Unidos
Fil: Rai, Priyamvada. Miami University. School Of Medicine; Estados Unidos
Fil: Monje, Paula. Miami University. School Of Medicine; Estados Unidos
Society for Neuroscience 46th Annual Meeting
San Diego
Estados Unidos
Society for Neuroscience - Materia
-
SCHWANN CELLS
SENESCENCE
PROLIFERATION
RETROVIRAL VECTORS - Nivel de accesibilidad
- acceso abierto
- Condiciones de uso
- https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
- Repositorio
- Institución
- Consejo Nacional de Investigaciones Científicas y Técnicas
- OAI Identificador
- oai:ri.conicet.gov.ar:11336/235392
Ver los metadatos del registro completo
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network_name_str |
CONICET Digital (CONICET) |
spelling |
Human Schwann cell senescence is not prevented by ectopic expression of human telomerase reverse transcriptaseAndersen, Natalia DeniseKuo, BlanchePiñero, Gonzalo MiguelRavelo, KristineRai, PriyamvadaMonje, PaulaSCHWANN CELLSSENESCENCEPROLIFERATIONRETROVIRAL VECTORShttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Isolated human Schwann cells (hSCs) typically become senescent and unresponsive to mitogenic factors with continued in vitro expansion. Data from our lab has shown that adult nerve-derived hSCs, in contrast to rodent (rat) SCs, inevitably stop proliferating and acquire a senescent phenotype characterized by high levels of senescence-associated (SA)-β galactosidase activity and morphological changes that include cell enlargement and appearance of multinucleated cells. As opposed to rodent SCs, hSC cultures consist of mixed populations of proliferating cells, senescent cells and cells at different stages of differentiation regardless of the nerve of origin and other donor-specific factors. RNA-seq analysis of representative cultures of hSCs did not reveal the presence of telomerase reverse transcriptase (TERT) mRNA while other TERT-related genes (e.g. TERF1, telomeric repeat binding factor, and TEP1, telomeraseassociated protein) were well-represented in the hSC transcriptome. In an attempt to overcome senescence, we used retroviral vectors and antibiotic selection to generate hSC lines ectopically expressing human (h)-TERT. For these experiments, highly proliferative, non-senescent, early passage hSC cultures were stably transduced with the retroviruses h-TERT-hygro or h-TERTpuro, each encoding the h-TERT gene along with hygromycin or puromycin resistance genes, respectively. Transduced hSC cultures from three different donors were selected and subjected to three rounds of expansion in medium containing chemical mitogens. Subsequently, the cultures were analyzed for their rate of proliferation by means of EdU incorporation assays and the acquisition of senescence by means of SA-β galactosidase activity assays in each round. We found that whereas ectopic h-TERT expression extended the lifespan of cultured hSCs when compared to non-infected or GFP-expressing cells, it was not sufficient to confer immortalization and overcome senescence. In sum, our results suggest that progression of the hSCs to a senescent state likely is stress-induced rather than dependent on replication-associated telomere shorteningFil: Andersen, Natalia Denise. Miami University; Estados Unidos. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Kuo, Blanche. Miami University; Estados UnidosFil: Piñero, Gonzalo Miguel. Miami University; Estados Unidos. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Ravelo, Kristine. Miami University; Estados UnidosFil: Rai, Priyamvada. Miami University. School Of Medicine; Estados UnidosFil: Monje, Paula. Miami University. School Of Medicine; Estados UnidosSociety for Neuroscience 46th Annual MeetingSan DiegoEstados UnidosSociety for NeuroscienceSociety for Neuroscience2016info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/conferenceObjectCongresoBookhttp://purl.org/coar/resource_type/c_5794info:ar-repo/semantics/documentoDeConferenciaapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/235392Human Schwann cell senescence is not prevented by ectopic expression of human telomerase reverse transcriptase; Society for Neuroscience 46th Annual Meeting; San Diego; Estados Unidos; 2016; 1-1CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://www.sfn.org/Annual-Meeting/Neuroscience-2016/Sessions-and-Events/Program/~/media/SfN/Documents/Annual%20Meeting/FinalProgram/NS2016/Full%20Abstract%20PDFs%202016/SfN16_Abstract%20PDFs%20%20Posters_3_Mon_PM.ashxinfo:eu-repo/semantics/altIdentifier/url/https://www.sfn.org/meetings/past-and-future-annual-meetingsInternacionalinfo:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T10:31:04Zoai:ri.conicet.gov.ar:11336/235392instacron: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:34982025-09-29 10:31:04.817CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse |
dc.title.none.fl_str_mv |
Human Schwann cell senescence is not prevented by ectopic expression of human telomerase reverse transcriptase |
title |
Human Schwann cell senescence is not prevented by ectopic expression of human telomerase reverse transcriptase |
spellingShingle |
Human Schwann cell senescence is not prevented by ectopic expression of human telomerase reverse transcriptase Andersen, Natalia Denise SCHWANN CELLS SENESCENCE PROLIFERATION RETROVIRAL VECTORS |
title_short |
Human Schwann cell senescence is not prevented by ectopic expression of human telomerase reverse transcriptase |
title_full |
Human Schwann cell senescence is not prevented by ectopic expression of human telomerase reverse transcriptase |
title_fullStr |
Human Schwann cell senescence is not prevented by ectopic expression of human telomerase reverse transcriptase |
title_full_unstemmed |
Human Schwann cell senescence is not prevented by ectopic expression of human telomerase reverse transcriptase |
title_sort |
Human Schwann cell senescence is not prevented by ectopic expression of human telomerase reverse transcriptase |
dc.creator.none.fl_str_mv |
Andersen, Natalia Denise Kuo, Blanche Piñero, Gonzalo Miguel Ravelo, Kristine Rai, Priyamvada Monje, Paula |
author |
Andersen, Natalia Denise |
author_facet |
Andersen, Natalia Denise Kuo, Blanche Piñero, Gonzalo Miguel Ravelo, Kristine Rai, Priyamvada Monje, Paula |
author_role |
author |
author2 |
Kuo, Blanche Piñero, Gonzalo Miguel Ravelo, Kristine Rai, Priyamvada Monje, Paula |
author2_role |
author author author author author |
dc.subject.none.fl_str_mv |
SCHWANN CELLS SENESCENCE PROLIFERATION RETROVIRAL VECTORS |
topic |
SCHWANN CELLS SENESCENCE PROLIFERATION RETROVIRAL VECTORS |
purl_subject.fl_str_mv |
https://purl.org/becyt/ford/1.6 https://purl.org/becyt/ford/1 |
dc.description.none.fl_txt_mv |
Isolated human Schwann cells (hSCs) typically become senescent and unresponsive to mitogenic factors with continued in vitro expansion. Data from our lab has shown that adult nerve-derived hSCs, in contrast to rodent (rat) SCs, inevitably stop proliferating and acquire a senescent phenotype characterized by high levels of senescence-associated (SA)-β galactosidase activity and morphological changes that include cell enlargement and appearance of multinucleated cells. As opposed to rodent SCs, hSC cultures consist of mixed populations of proliferating cells, senescent cells and cells at different stages of differentiation regardless of the nerve of origin and other donor-specific factors. RNA-seq analysis of representative cultures of hSCs did not reveal the presence of telomerase reverse transcriptase (TERT) mRNA while other TERT-related genes (e.g. TERF1, telomeric repeat binding factor, and TEP1, telomeraseassociated protein) were well-represented in the hSC transcriptome. In an attempt to overcome senescence, we used retroviral vectors and antibiotic selection to generate hSC lines ectopically expressing human (h)-TERT. For these experiments, highly proliferative, non-senescent, early passage hSC cultures were stably transduced with the retroviruses h-TERT-hygro or h-TERTpuro, each encoding the h-TERT gene along with hygromycin or puromycin resistance genes, respectively. Transduced hSC cultures from three different donors were selected and subjected to three rounds of expansion in medium containing chemical mitogens. Subsequently, the cultures were analyzed for their rate of proliferation by means of EdU incorporation assays and the acquisition of senescence by means of SA-β galactosidase activity assays in each round. We found that whereas ectopic h-TERT expression extended the lifespan of cultured hSCs when compared to non-infected or GFP-expressing cells, it was not sufficient to confer immortalization and overcome senescence. In sum, our results suggest that progression of the hSCs to a senescent state likely is stress-induced rather than dependent on replication-associated telomere shortening Fil: Andersen, Natalia Denise. Miami University; Estados Unidos. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Kuo, Blanche. Miami University; Estados Unidos Fil: Piñero, Gonzalo Miguel. Miami University; Estados Unidos. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Ravelo, Kristine. Miami University; Estados Unidos Fil: Rai, Priyamvada. Miami University. School Of Medicine; Estados Unidos Fil: Monje, Paula. Miami University. School Of Medicine; Estados Unidos Society for Neuroscience 46th Annual Meeting San Diego Estados Unidos Society for Neuroscience |
description |
Isolated human Schwann cells (hSCs) typically become senescent and unresponsive to mitogenic factors with continued in vitro expansion. Data from our lab has shown that adult nerve-derived hSCs, in contrast to rodent (rat) SCs, inevitably stop proliferating and acquire a senescent phenotype characterized by high levels of senescence-associated (SA)-β galactosidase activity and morphological changes that include cell enlargement and appearance of multinucleated cells. As opposed to rodent SCs, hSC cultures consist of mixed populations of proliferating cells, senescent cells and cells at different stages of differentiation regardless of the nerve of origin and other donor-specific factors. RNA-seq analysis of representative cultures of hSCs did not reveal the presence of telomerase reverse transcriptase (TERT) mRNA while other TERT-related genes (e.g. TERF1, telomeric repeat binding factor, and TEP1, telomeraseassociated protein) were well-represented in the hSC transcriptome. In an attempt to overcome senescence, we used retroviral vectors and antibiotic selection to generate hSC lines ectopically expressing human (h)-TERT. For these experiments, highly proliferative, non-senescent, early passage hSC cultures were stably transduced with the retroviruses h-TERT-hygro or h-TERTpuro, each encoding the h-TERT gene along with hygromycin or puromycin resistance genes, respectively. Transduced hSC cultures from three different donors were selected and subjected to three rounds of expansion in medium containing chemical mitogens. Subsequently, the cultures were analyzed for their rate of proliferation by means of EdU incorporation assays and the acquisition of senescence by means of SA-β galactosidase activity assays in each round. We found that whereas ectopic h-TERT expression extended the lifespan of cultured hSCs when compared to non-infected or GFP-expressing cells, it was not sufficient to confer immortalization and overcome senescence. In sum, our results suggest that progression of the hSCs to a senescent state likely is stress-induced rather than dependent on replication-associated telomere shortening |
publishDate |
2016 |
dc.date.none.fl_str_mv |
2016 |
dc.type.none.fl_str_mv |
info:eu-repo/semantics/publishedVersion info:eu-repo/semantics/conferenceObject Congreso Book http://purl.org/coar/resource_type/c_5794 info:ar-repo/semantics/documentoDeConferencia |
status_str |
publishedVersion |
format |
conferenceObject |
dc.identifier.none.fl_str_mv |
http://hdl.handle.net/11336/235392 Human Schwann cell senescence is not prevented by ectopic expression of human telomerase reverse transcriptase; Society for Neuroscience 46th Annual Meeting; San Diego; Estados Unidos; 2016; 1-1 CONICET Digital CONICET |
url |
http://hdl.handle.net/11336/235392 |
identifier_str_mv |
Human Schwann cell senescence is not prevented by ectopic expression of human telomerase reverse transcriptase; Society for Neuroscience 46th Annual Meeting; San Diego; Estados Unidos; 2016; 1-1 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.sfn.org/Annual-Meeting/Neuroscience-2016/Sessions-and-Events/Program/~/media/SfN/Documents/Annual%20Meeting/FinalProgram/NS2016/Full%20Abstract%20PDFs%202016/SfN16_Abstract%20PDFs%20%20Posters_3_Mon_PM.ashx info:eu-repo/semantics/altIdentifier/url/https://www.sfn.org/meetings/past-and-future-annual-meetings |
dc.rights.none.fl_str_mv |
info:eu-repo/semantics/openAccess https://creativecommons.org/licenses/by-nc-sa/2.5/ar/ |
eu_rights_str_mv |
openAccess |
rights_invalid_str_mv |
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/ |
dc.format.none.fl_str_mv |
application/pdf application/pdf application/pdf |
dc.coverage.none.fl_str_mv |
Internacional |
dc.publisher.none.fl_str_mv |
Society for Neuroscience |
publisher.none.fl_str_mv |
Society for Neuroscience |
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Consejo Nacional de Investigaciones Científicas y Técnicas |
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CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas |
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dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar |
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