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
- Institución
- Universidad Nacional de La Plata
- OAI Identificador
- oai:sedici.unlp.edu.ar:10915/154700
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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 |
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article |
status_str |
publishedVersion |
dc.identifier.none.fl_str_mv |
http://sedici.unlp.edu.ar/handle/10915/154700 |
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http://sedici.unlp.edu.ar/handle/10915/154700 |
dc.language.none.fl_str_mv |
eng |
language |
eng |
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info:eu-repo/semantics/altIdentifier/issn/1432-0878 info:eu-repo/semantics/altIdentifier/doi/10.1007/s00441-017-2608-x |
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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) |
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openAccess |
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http://creativecommons.org/licenses/by-nc-sa/4.0/ Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0) |
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