Extended cold storage of cultured hepatocytes impairs endocytic uptake during normothermic rewarming

Autores
Hovanyecz, P.; Guibert, Edgardo Elvio; Pellegrino, Jose Manuel; Rodriguez, Joaquin Valentin; Sigot, Valeria
Año de publicación
2013
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
During hypothermic preservation of cells (0-4°C), metab¬olism is diminished and energy-dependent transport processes are arrested. The effect of hypothermic preservation of hepatocytes in endocytic transport following rewarming has not been previously reported. We evaluated the uptake of EGF (Epidermal Growth Factor) ligand conjugated to fluorescent Quantum Dots (QDs) probes in rat hepatocytes after 24 and 72 h cold storage in University of Wisconsin (UW) solution at 4°C. QDs uptake was visualized during rewarming to 37°C under air or, in a second approach, at the end of rewarming under 5% CO2. After 24 h in UW solution, QDs were internalized under both rewarming conditions similar to non-preserved hepatocytes and cells maintained a normal cytoskeleton distribution. However, in hepatocytes preserved 72 h none of the cells internalized QDs, which remained bound to the membranes. After rewarming, this group showed diminished actin staining while viability was maintained at ~70%. Our results present evidence that, hypothermic preservation for 72 h in UW solution at 4°C does not prevent EGFR (Epidermal Growth Factor Receptor) activation but irreversibly impairs endocytic uptake upon EGF stimulation; presumably due to actin cytoskeleton disassembling. Our approach can be applied on other membrane receptor systems and with other hypothermic preservation solutions to understand the effect of cooling in endocytic transport and to determine the optimal cold storage period.
Fil: Hovanyecz, P.. Universidad Nacional de Rosario. Secretaria de Ciencia y Tecnica. Centro Binacional de Investigación en Criobiologia Clinica y Aplicada; Argentina
Fil: Guibert, Edgardo Elvio. Universidad Nacional de Rosario. Secretaria de Ciencia y Tecnica. Centro Binacional de Investigación en Criobiologia Clinica y Aplicada; Argentina. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas; Argentina
Fil: Pellegrino, Jose Manuel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Rosario. Instituto de Fisiología Experimental (i); Argentina
Fil: Rodriguez, Joaquin Valentin. Universidad Nacional de Rosario. Secretaria de Ciencia y Tecnica. Centro Binacional de Investigación en Criobiologia Clinica y Aplicada; Argentina
Fil: Sigot, Valeria. Universidad Nacional de Rosario. Secretaria de Ciencia y Tecnica. Centro Binacional de Investigación en Criobiologia Clinica y Aplicada; Argentina
Materia
Hypothermic preservation
Receptor-mediated endocytosis
Cultured rat hepatocytes
Epidermal growth factor receptor
Quantum dots
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/7375
Acceder
id CONICETDig_35e94fa71a36f7703eb4c6310d41dfb4
oai_identifier_str oai:ri.conicet.gov.ar:11336/7375
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Extended cold storage of cultured hepatocytes impairs endocytic uptake during normothermic rewarmingHovanyecz, P.Guibert, Edgardo ElvioPellegrino, Jose ManuelRodriguez, Joaquin ValentinSigot, ValeriaHypothermic preservationReceptor-mediated endocytosisCultured rat hepatocytesEpidermal growth factor receptorQuantum dotshttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1During hypothermic preservation of cells (0-4°C), metab¬olism is diminished and energy-dependent transport processes are arrested. The effect of hypothermic preservation of hepatocytes in endocytic transport following rewarming has not been previously reported. We evaluated the uptake of EGF (Epidermal Growth Factor) ligand conjugated to fluorescent Quantum Dots (QDs) probes in rat hepatocytes after 24 and 72 h cold storage in University of Wisconsin (UW) solution at 4°C. QDs uptake was visualized during rewarming to 37°C under air or, in a second approach, at the end of rewarming under 5% CO2. After 24 h in UW solution, QDs were internalized under both rewarming conditions similar to non-preserved hepatocytes and cells maintained a normal cytoskeleton distribution. However, in hepatocytes preserved 72 h none of the cells internalized QDs, which remained bound to the membranes. After rewarming, this group showed diminished actin staining while viability was maintained at ~70%. Our results present evidence that, hypothermic preservation for 72 h in UW solution at 4°C does not prevent EGFR (Epidermal Growth Factor Receptor) activation but irreversibly impairs endocytic uptake upon EGF stimulation; presumably due to actin cytoskeleton disassembling. Our approach can be applied on other membrane receptor systems and with other hypothermic preservation solutions to understand the effect of cooling in endocytic transport and to determine the optimal cold storage period.Fil: Hovanyecz, P.. Universidad Nacional de Rosario. Secretaria de Ciencia y Tecnica. Centro Binacional de Investigación en Criobiologia Clinica y Aplicada; ArgentinaFil: Guibert, Edgardo Elvio. Universidad Nacional de Rosario. Secretaria de Ciencia y Tecnica. Centro Binacional de Investigación en Criobiologia Clinica y Aplicada; Argentina. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas; ArgentinaFil: Pellegrino, Jose Manuel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Rosario. Instituto de Fisiología Experimental (i); ArgentinaFil: Rodriguez, Joaquin Valentin. Universidad Nacional de Rosario. Secretaria de Ciencia y Tecnica. Centro Binacional de Investigación en Criobiologia Clinica y Aplicada; ArgentinaFil: Sigot, Valeria. Universidad Nacional de Rosario. Secretaria de Ciencia y Tecnica. Centro Binacional de Investigación en Criobiologia Clinica y Aplicada; ArgentinaElsevier2013-04info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfapplication/pdfapplication/pdfapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/7375Hovanyecz, P.; Guibert, Edgardo Elvio; Pellegrino, Jose Manuel; Rodriguez, Joaquin Valentin; Sigot, Valeria; Extended cold storage of cultured hepatocytes impairs endocytic uptake during normothermic rewarming; Elsevier; Cryobiology; 66; 2; 4-2013; 112-1200011-2240enginfo:eu-repo/semantics/altIdentifier/url/http://www.sciencedirect.com/science/article/pii/S001122401200274Xinfo:eu-repo/semantics/altIdentifier/doi/10.1016/j.cryobiol.2012.12.004info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-nd/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T09:51:43Zoai:ri.conicet.gov.ar:11336/7375instacron: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 09:51:43.315CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Extended cold storage of cultured hepatocytes impairs endocytic uptake during normothermic rewarming
title Extended cold storage of cultured hepatocytes impairs endocytic uptake during normothermic rewarming
spellingShingle Extended cold storage of cultured hepatocytes impairs endocytic uptake during normothermic rewarming
Hovanyecz, P.
Hypothermic preservation
Receptor-mediated endocytosis
Cultured rat hepatocytes
Epidermal growth factor receptor
Quantum dots
title_short Extended cold storage of cultured hepatocytes impairs endocytic uptake during normothermic rewarming
title_full Extended cold storage of cultured hepatocytes impairs endocytic uptake during normothermic rewarming
title_fullStr Extended cold storage of cultured hepatocytes impairs endocytic uptake during normothermic rewarming
title_full_unstemmed Extended cold storage of cultured hepatocytes impairs endocytic uptake during normothermic rewarming
title_sort Extended cold storage of cultured hepatocytes impairs endocytic uptake during normothermic rewarming
dc.creator.none.fl_str_mv Hovanyecz, P.
Guibert, Edgardo Elvio
Pellegrino, Jose Manuel
Rodriguez, Joaquin Valentin
Sigot, Valeria
author Hovanyecz, P.
author_facet Hovanyecz, P.
Guibert, Edgardo Elvio
Pellegrino, Jose Manuel
Rodriguez, Joaquin Valentin
Sigot, Valeria
author_role author
author2 Guibert, Edgardo Elvio
Pellegrino, Jose Manuel
Rodriguez, Joaquin Valentin
Sigot, Valeria
author2_role author
author
author
author
dc.subject.none.fl_str_mv Hypothermic preservation
Receptor-mediated endocytosis
Cultured rat hepatocytes
Epidermal growth factor receptor
Quantum dots
topic Hypothermic preservation
Receptor-mediated endocytosis
Cultured rat hepatocytes
Epidermal growth factor receptor
Quantum dots
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv During hypothermic preservation of cells (0-4°C), metab¬olism is diminished and energy-dependent transport processes are arrested. The effect of hypothermic preservation of hepatocytes in endocytic transport following rewarming has not been previously reported. We evaluated the uptake of EGF (Epidermal Growth Factor) ligand conjugated to fluorescent Quantum Dots (QDs) probes in rat hepatocytes after 24 and 72 h cold storage in University of Wisconsin (UW) solution at 4°C. QDs uptake was visualized during rewarming to 37°C under air or, in a second approach, at the end of rewarming under 5% CO2. After 24 h in UW solution, QDs were internalized under both rewarming conditions similar to non-preserved hepatocytes and cells maintained a normal cytoskeleton distribution. However, in hepatocytes preserved 72 h none of the cells internalized QDs, which remained bound to the membranes. After rewarming, this group showed diminished actin staining while viability was maintained at ~70%. Our results present evidence that, hypothermic preservation for 72 h in UW solution at 4°C does not prevent EGFR (Epidermal Growth Factor Receptor) activation but irreversibly impairs endocytic uptake upon EGF stimulation; presumably due to actin cytoskeleton disassembling. Our approach can be applied on other membrane receptor systems and with other hypothermic preservation solutions to understand the effect of cooling in endocytic transport and to determine the optimal cold storage period.
Fil: Hovanyecz, P.. Universidad Nacional de Rosario. Secretaria de Ciencia y Tecnica. Centro Binacional de Investigación en Criobiologia Clinica y Aplicada; Argentina
Fil: Guibert, Edgardo Elvio. Universidad Nacional de Rosario. Secretaria de Ciencia y Tecnica. Centro Binacional de Investigación en Criobiologia Clinica y Aplicada; Argentina. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas; Argentina
Fil: Pellegrino, Jose Manuel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Rosario. Instituto de Fisiología Experimental (i); Argentina
Fil: Rodriguez, Joaquin Valentin. Universidad Nacional de Rosario. Secretaria de Ciencia y Tecnica. Centro Binacional de Investigación en Criobiologia Clinica y Aplicada; Argentina
Fil: Sigot, Valeria. Universidad Nacional de Rosario. Secretaria de Ciencia y Tecnica. Centro Binacional de Investigación en Criobiologia Clinica y Aplicada; Argentina
description During hypothermic preservation of cells (0-4°C), metab¬olism is diminished and energy-dependent transport processes are arrested. The effect of hypothermic preservation of hepatocytes in endocytic transport following rewarming has not been previously reported. We evaluated the uptake of EGF (Epidermal Growth Factor) ligand conjugated to fluorescent Quantum Dots (QDs) probes in rat hepatocytes after 24 and 72 h cold storage in University of Wisconsin (UW) solution at 4°C. QDs uptake was visualized during rewarming to 37°C under air or, in a second approach, at the end of rewarming under 5% CO2. After 24 h in UW solution, QDs were internalized under both rewarming conditions similar to non-preserved hepatocytes and cells maintained a normal cytoskeleton distribution. However, in hepatocytes preserved 72 h none of the cells internalized QDs, which remained bound to the membranes. After rewarming, this group showed diminished actin staining while viability was maintained at ~70%. Our results present evidence that, hypothermic preservation for 72 h in UW solution at 4°C does not prevent EGFR (Epidermal Growth Factor Receptor) activation but irreversibly impairs endocytic uptake upon EGF stimulation; presumably due to actin cytoskeleton disassembling. Our approach can be applied on other membrane receptor systems and with other hypothermic preservation solutions to understand the effect of cooling in endocytic transport and to determine the optimal cold storage period.
publishDate 2013
dc.date.none.fl_str_mv 2013-04
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/11336/7375
Hovanyecz, P.; Guibert, Edgardo Elvio; Pellegrino, Jose Manuel; Rodriguez, Joaquin Valentin; Sigot, Valeria; Extended cold storage of cultured hepatocytes impairs endocytic uptake during normothermic rewarming; Elsevier; Cryobiology; 66; 2; 4-2013; 112-120
0011-2240
url http://hdl.handle.net/11336/7375
identifier_str_mv Hovanyecz, P.; Guibert, Edgardo Elvio; Pellegrino, Jose Manuel; Rodriguez, Joaquin Valentin; Sigot, Valeria; Extended cold storage of cultured hepatocytes impairs endocytic uptake during normothermic rewarming; Elsevier; Cryobiology; 66; 2; 4-2013; 112-120
0011-2240
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/http://www.sciencedirect.com/science/article/pii/S001122401200274X
info:eu-repo/semantics/altIdentifier/doi/10.1016/j.cryobiol.2012.12.004
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
application/pdf
application/pdf
application/pdf
application/pdf
application/pdf
application/pdf
dc.publisher.none.fl_str_mv Elsevier
publisher.none.fl_str_mv Elsevier
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
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score 13.070432

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