A simple strategy for culturing morphologically-conserved rat hypothalamic tanycytes

Autores
De Francesco, Pablo Nicolás; Castrogiovanni, Daniel Cayetano; Uriarte, Maia; Frassa, María Victoria; Agosti, Francina; Raingo, Jesica; Perelló, Mario Carlos
Año de publicación
2017
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Hypothalamic tanycytes are specialized bipolar ependymal cells that line the floor of the third ventricle. Given their strategic location, tanycytes are believed to play several key functions including being a selective barrier and controlling the amount of hypothalamic-derived factors reaching the anterior pituitary. The in vitro culture of these cells has proved to be difficult. Here, we report an improved method for the generation of primary cultures of rat hypothalamic tanycytes. Ependymal cultures were derived from tissue dissected out of the median eminence region of 10-day-old rats and cultured in a chemically defined medium containing DMEM:F12, serum albumin, insulin, transferrin and the antibiotic gentamycin. After 7 days in vitro, -30% of the cultured cells exhibited morphological features of tanycytes as observed by phase contrast or scanning electron microscopy. Tanycyte-like cells were strongly immuno-reactive for vimentin and dopamine-cAMP-regulated phospho-protein (DARPP-32) and weakly immune-reactive for glial fibrillary acidic protein. Tanycyte-like cells displayed a stable negative resting plasma membrane potential and failed to show spiking properties in response to current injections. When exposed to fluorescent beads in the culture medium, tanycyte-like cells exhibited a robust endocytosis. Thus, the present method effectively yields cultures containing tanycyte-like cells that resemble in vivo tanycytes in terms of morphologic features and molecular markers as well as electrical and endocytic activity. To our knowledge, this is the first protocol that allows the culturing of tanycyte-like cells that can be individually identified and that conserve the morphology of tanycytes in then- natural physiological environment.
Instituto Multidisciplinario de Biología Celular
Comisión de Investigaciones Científicas de la provincia de Buenos Aires
Materia
Biología
Cell culture
Ependymal cells
Hypothalamus
Median eminence
Glia
Nivel de accesibilidad
acceso abierto
Condiciones de uso
http://creativecommons.org/licenses/by-nc-sa/4.0/
Repositorio
SEDICI (UNLP)
Institución
Universidad Nacional de La Plata
OAI Identificador
oai:sedici.unlp.edu.ar:10915/154700

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oai_identifier_str oai:sedici.unlp.edu.ar:10915/154700
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repository_id_str 1329
network_name_str SEDICI (UNLP)
spelling A simple strategy for culturing morphologically-conserved rat hypothalamic tanycytesDe Francesco, Pablo NicolásCastrogiovanni, Daniel CayetanoUriarte, MaiaFrassa, María VictoriaAgosti, FrancinaRaingo, JesicaPerelló, Mario CarlosBiologíaCell cultureEpendymal cellsHypothalamusMedian eminenceGliaHypothalamic tanycytes are specialized bipolar ependymal cells that line the floor of the third ventricle. Given their strategic location, tanycytes are believed to play several key functions including being a selective barrier and controlling the amount of hypothalamic-derived factors reaching the anterior pituitary. The in vitro culture of these cells has proved to be difficult. Here, we report an improved method for the generation of primary cultures of rat hypothalamic tanycytes. Ependymal cultures were derived from tissue dissected out of the median eminence region of 10-day-old rats and cultured in a chemically defined medium containing DMEM:F12, serum albumin, insulin, transferrin and the antibiotic gentamycin. After 7 days in vitro, -30% of the cultured cells exhibited morphological features of tanycytes as observed by phase contrast or scanning electron microscopy. Tanycyte-like cells were strongly immuno-reactive for vimentin and dopamine-cAMP-regulated phospho-protein (DARPP-32) and weakly immune-reactive for glial fibrillary acidic protein. Tanycyte-like cells displayed a stable negative resting plasma membrane potential and failed to show spiking properties in response to current injections. When exposed to fluorescent beads in the culture medium, tanycyte-like cells exhibited a robust endocytosis. Thus, the present method effectively yields cultures containing tanycyte-like cells that resemble in vivo tanycytes in terms of morphologic features and molecular markers as well as electrical and endocytic activity. To our knowledge, this is the first protocol that allows the culturing of tanycyte-like cells that can be individually identified and that conserve the morphology of tanycytes in then- natural physiological environment.Instituto Multidisciplinario de Biología CelularComisión de Investigaciones Científicas de la provincia de Buenos Aires2017info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionArticulohttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdf369-380http://sedici.unlp.edu.ar/handle/10915/154700enginfo:eu-repo/semantics/altIdentifier/issn/1432-0878info:eu-repo/semantics/altIdentifier/doi/10.1007/s00441-017-2608-xinfo:eu-repo/semantics/openAccesshttp://creativecommons.org/licenses/by-nc-sa/4.0/Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0)reponame:SEDICI (UNLP)instname:Universidad Nacional de La Platainstacron:UNLP2025-09-29T11:40:05Zoai:sedici.unlp.edu.ar:10915/154700Institucionalhttp://sedici.unlp.edu.ar/Universidad públicaNo correspondehttp://sedici.unlp.edu.ar/oai/snrdalira@sedici.unlp.edu.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:13292025-09-29 11:40:05.738SEDICI (UNLP) - Universidad Nacional de La Platafalse
dc.title.none.fl_str_mv A simple strategy for culturing morphologically-conserved rat hypothalamic tanycytes
title A simple strategy for culturing morphologically-conserved rat hypothalamic tanycytes
spellingShingle A simple strategy for culturing morphologically-conserved rat hypothalamic tanycytes
De Francesco, Pablo Nicolás
Biología
Cell culture
Ependymal cells
Hypothalamus
Median eminence
Glia
title_short A simple strategy for culturing morphologically-conserved rat hypothalamic tanycytes
title_full A simple strategy for culturing morphologically-conserved rat hypothalamic tanycytes
title_fullStr A simple strategy for culturing morphologically-conserved rat hypothalamic tanycytes
title_full_unstemmed A simple strategy for culturing morphologically-conserved rat hypothalamic tanycytes
title_sort A simple strategy for culturing morphologically-conserved rat hypothalamic tanycytes
dc.creator.none.fl_str_mv De Francesco, Pablo Nicolás
Castrogiovanni, Daniel Cayetano
Uriarte, Maia
Frassa, María Victoria
Agosti, Francina
Raingo, Jesica
Perelló, Mario Carlos
author De Francesco, Pablo Nicolás
author_facet De Francesco, Pablo Nicolás
Castrogiovanni, Daniel Cayetano
Uriarte, Maia
Frassa, María Victoria
Agosti, Francina
Raingo, Jesica
Perelló, Mario Carlos
author_role author
author2 Castrogiovanni, Daniel Cayetano
Uriarte, Maia
Frassa, María Victoria
Agosti, Francina
Raingo, Jesica
Perelló, Mario Carlos
author2_role author
author
author
author
author
author
dc.subject.none.fl_str_mv Biología
Cell culture
Ependymal cells
Hypothalamus
Median eminence
Glia
topic Biología
Cell culture
Ependymal cells
Hypothalamus
Median eminence
Glia
dc.description.none.fl_txt_mv Hypothalamic tanycytes are specialized bipolar ependymal cells that line the floor of the third ventricle. Given their strategic location, tanycytes are believed to play several key functions including being a selective barrier and controlling the amount of hypothalamic-derived factors reaching the anterior pituitary. The in vitro culture of these cells has proved to be difficult. Here, we report an improved method for the generation of primary cultures of rat hypothalamic tanycytes. Ependymal cultures were derived from tissue dissected out of the median eminence region of 10-day-old rats and cultured in a chemically defined medium containing DMEM:F12, serum albumin, insulin, transferrin and the antibiotic gentamycin. After 7 days in vitro, -30% of the cultured cells exhibited morphological features of tanycytes as observed by phase contrast or scanning electron microscopy. Tanycyte-like cells were strongly immuno-reactive for vimentin and dopamine-cAMP-regulated phospho-protein (DARPP-32) and weakly immune-reactive for glial fibrillary acidic protein. Tanycyte-like cells displayed a stable negative resting plasma membrane potential and failed to show spiking properties in response to current injections. When exposed to fluorescent beads in the culture medium, tanycyte-like cells exhibited a robust endocytosis. Thus, the present method effectively yields cultures containing tanycyte-like cells that resemble in vivo tanycytes in terms of morphologic features and molecular markers as well as electrical and endocytic activity. To our knowledge, this is the first protocol that allows the culturing of tanycyte-like cells that can be individually identified and that conserve the morphology of tanycytes in then- natural physiological environment.
Instituto Multidisciplinario de Biología Celular
Comisión de Investigaciones Científicas de la provincia de Buenos Aires
description Hypothalamic tanycytes are specialized bipolar ependymal cells that line the floor of the third ventricle. Given their strategic location, tanycytes are believed to play several key functions including being a selective barrier and controlling the amount of hypothalamic-derived factors reaching the anterior pituitary. The in vitro culture of these cells has proved to be difficult. Here, we report an improved method for the generation of primary cultures of rat hypothalamic tanycytes. Ependymal cultures were derived from tissue dissected out of the median eminence region of 10-day-old rats and cultured in a chemically defined medium containing DMEM:F12, serum albumin, insulin, transferrin and the antibiotic gentamycin. After 7 days in vitro, -30% of the cultured cells exhibited morphological features of tanycytes as observed by phase contrast or scanning electron microscopy. Tanycyte-like cells were strongly immuno-reactive for vimentin and dopamine-cAMP-regulated phospho-protein (DARPP-32) and weakly immune-reactive for glial fibrillary acidic protein. Tanycyte-like cells displayed a stable negative resting plasma membrane potential and failed to show spiking properties in response to current injections. When exposed to fluorescent beads in the culture medium, tanycyte-like cells exhibited a robust endocytosis. Thus, the present method effectively yields cultures containing tanycyte-like cells that resemble in vivo tanycytes in terms of morphologic features and molecular markers as well as electrical and endocytic activity. To our knowledge, this is the first protocol that allows the culturing of tanycyte-like cells that can be individually identified and that conserve the morphology of tanycytes in then- natural physiological environment.
publishDate 2017
dc.date.none.fl_str_mv 2017
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
Articulo
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://sedici.unlp.edu.ar/handle/10915/154700
url http://sedici.unlp.edu.ar/handle/10915/154700
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/issn/1432-0878
info:eu-repo/semantics/altIdentifier/doi/10.1007/s00441-017-2608-x
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by-nc-sa/4.0/
Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0)
eu_rights_str_mv openAccess
rights_invalid_str_mv http://creativecommons.org/licenses/by-nc-sa/4.0/
Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0)
dc.format.none.fl_str_mv application/pdf
369-380
dc.source.none.fl_str_mv reponame:SEDICI (UNLP)
instname:Universidad Nacional de La Plata
instacron:UNLP
reponame_str SEDICI (UNLP)
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instname_str Universidad Nacional de La Plata
instacron_str UNLP
institution UNLP
repository.name.fl_str_mv SEDICI (UNLP) - Universidad Nacional de La Plata
repository.mail.fl_str_mv alira@sedici.unlp.edu.ar
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