Co-localization of nestin and insulin and expression of islet cell markers in long-term human pancreatic nestin-positive cell cultures

Detalhes bibliográficos
Autor(a) principal: Maria-Engler, Sylvia Stucchi
Data de Publicação: 2004
Outros Autores: Correa-Giannella, Maria Lucia C, Labriola, Leticia, Krogh, Karin, Colin, Christian, Lojudice, Fernando Henrique, Aita, Carlos Alberto Mayora, Oliveira, Elizabeth Maria Costa de, Correa, Tatiana C Silveira, Silva, Irenice Cairo da, Genzini, Tercio, Miranda, Marcelo Perosa de, Noronha, Irene Lourdes, Vilela, Luciano, Coimbra, Cassio Negro, Mortara, Renato A. [UNIFESP], Guia, Marcos Mares, Eliaschewitz, Freddy Goldenberg, Sogayar, Mari Cleide
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Institucional da UNIFESP
Texto Completo: http://dx.doi.org/10.1677/joe.1.05703
http://repositorio.unifesp.br/handle/11600/28032
Resumo: Strategies to differentiate progenitor cells into beta cells in vitro have been considered as an alternative to increase beta cell availability prior to transplantation. It has recently been suggested that nestin-positive cells could be multipotential stern cells capable of expressing endocrine markers upon specific stimulation; however, this issue still remains controversial. Here, we characterized short- and long-term islet cell cultures derived from three different human islet preparations. with respect to expression of nestin and islet cell markers, using confocal microscopy and semi-quantitative PT-PCR. the number of nestin-positive cells was found to be strikingly high in long-term cultures. in addition, a large proportion (49.7%) of these nestin-positive cells, present in long-term culture, are shown to be proliferative, as judged by BrdU incorporation. the proportion of insulin-positive cells was found to be high in short-term (up to 28 days) cultures and declined thereafter, when cells were maintained in the presence of 10% serum, concomitantly with the decrease in insulin and PDX-1 expression. Interestingly, insulin and nestin co-expression was observed as a rare event in a small proportion of cells present in freshly isolated human islets as well as in purified islet cells cultured in vitro for long periods of time. in addition, upon long-term subculturing of nestin-positive cells in 10% serum, we observed reappearance of insulin expression at the mRNA level; when these cultures were shifted to 1% serum for a month, expression of insulin, glucagon and somatostatin was also detected, indicating that manipulating the culture conditions can be used to modulate the nestin-positive cell's fate. Attempts to induce cell differentiation by plating nestin-positive cells onto Matrigel revealed that these cells tend to aggregate to form islet-like clusters, but this is not sufficient to increase insulin expression upon short-term culture. Our data corroborate previous findings indicating that, at least ill vitro, nestin-positive cells may undergo the early stages of differentiation to an islet cell phenotype and that long-term cultures of nestin-positive human islet cells may be considered as a potential source of precursor cells to generate fully differentiated/functional beta cells.
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spelling Co-localization of nestin and insulin and expression of islet cell markers in long-term human pancreatic nestin-positive cell culturesStrategies to differentiate progenitor cells into beta cells in vitro have been considered as an alternative to increase beta cell availability prior to transplantation. It has recently been suggested that nestin-positive cells could be multipotential stern cells capable of expressing endocrine markers upon specific stimulation; however, this issue still remains controversial. Here, we characterized short- and long-term islet cell cultures derived from three different human islet preparations. with respect to expression of nestin and islet cell markers, using confocal microscopy and semi-quantitative PT-PCR. the number of nestin-positive cells was found to be strikingly high in long-term cultures. in addition, a large proportion (49.7%) of these nestin-positive cells, present in long-term culture, are shown to be proliferative, as judged by BrdU incorporation. the proportion of insulin-positive cells was found to be high in short-term (up to 28 days) cultures and declined thereafter, when cells were maintained in the presence of 10% serum, concomitantly with the decrease in insulin and PDX-1 expression. Interestingly, insulin and nestin co-expression was observed as a rare event in a small proportion of cells present in freshly isolated human islets as well as in purified islet cells cultured in vitro for long periods of time. in addition, upon long-term subculturing of nestin-positive cells in 10% serum, we observed reappearance of insulin expression at the mRNA level; when these cultures were shifted to 1% serum for a month, expression of insulin, glucagon and somatostatin was also detected, indicating that manipulating the culture conditions can be used to modulate the nestin-positive cell's fate. Attempts to induce cell differentiation by plating nestin-positive cells onto Matrigel revealed that these cells tend to aggregate to form islet-like clusters, but this is not sufficient to increase insulin expression upon short-term culture. Our data corroborate previous findings indicating that, at least ill vitro, nestin-positive cells may undergo the early stages of differentiation to an islet cell phenotype and that long-term cultures of nestin-positive human islet cells may be considered as a potential source of precursor cells to generate fully differentiated/functional beta cells.Univ São Paulo, Inst Quim, Dept Bioquim, BR-05513970 São Paulo, BrazilUniv São Paulo, Fac Med, Hosp Clin, Lab Nutr Humana & Doencas Metab LIM25, São Paulo, BrazilAlbert Einstein Hosp, São Paulo, BrazilBiomm SA, Montes Claros, MG, BrazilUNIFESP, Escola Paulista Med, Dept Micro Imuno & Parasitol, São Paulo, BrazilUNIFESP, Escola Paulista Med, Dept Micro Imuno & Parasitol, São Paulo, BrazilWeb of ScienceSoc EndocrinologyUniversidade de São Paulo (USP)Albert Einstein HospBiomm SAUniversidade Federal de São Paulo (UNIFESP)Maria-Engler, Sylvia StucchiCorrea-Giannella, Maria Lucia CLabriola, LeticiaKrogh, KarinColin, ChristianLojudice, Fernando HenriqueAita, Carlos Alberto MayoraOliveira, Elizabeth Maria Costa deCorrea, Tatiana C SilveiraSilva, Irenice Cairo daGenzini, TercioMiranda, Marcelo Perosa deNoronha, Irene LourdesVilela, LucianoCoimbra, Cassio NegroMortara, Renato A. [UNIFESP]Guia, Marcos MaresEliaschewitz, Freddy GoldenbergSogayar, Mari Cleide2016-01-24T12:37:30Z2016-01-24T12:37:30Z2004-12-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersion455-467http://dx.doi.org/10.1677/joe.1.05703Journal of Endocrinology. Bristol: Soc Endocrinology, v. 183, n. 3, p. 455-467, 2004.10.1677/joe.1.057030022-0795http://repositorio.unifesp.br/handle/11600/28032WOS:000225947100003engJournal of Endocrinologyinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da UNIFESPinstname:Universidade Federal de São Paulo (UNIFESP)instacron:UNIFESP2022-02-08T11:55:00Zoai:repositorio.unifesp.br/:11600/28032Repositório InstitucionalPUBhttp://www.repositorio.unifesp.br/oai/requestbiblioteca.csp@unifesp.bropendoar:34652022-02-08T11:55Repositório Institucional da UNIFESP - Universidade Federal de São Paulo (UNIFESP)false
dc.title.none.fl_str_mv Co-localization of nestin and insulin and expression of islet cell markers in long-term human pancreatic nestin-positive cell cultures
title Co-localization of nestin and insulin and expression of islet cell markers in long-term human pancreatic nestin-positive cell cultures
spellingShingle Co-localization of nestin and insulin and expression of islet cell markers in long-term human pancreatic nestin-positive cell cultures
Maria-Engler, Sylvia Stucchi
title_short Co-localization of nestin and insulin and expression of islet cell markers in long-term human pancreatic nestin-positive cell cultures
title_full Co-localization of nestin and insulin and expression of islet cell markers in long-term human pancreatic nestin-positive cell cultures
title_fullStr Co-localization of nestin and insulin and expression of islet cell markers in long-term human pancreatic nestin-positive cell cultures
title_full_unstemmed Co-localization of nestin and insulin and expression of islet cell markers in long-term human pancreatic nestin-positive cell cultures
title_sort Co-localization of nestin and insulin and expression of islet cell markers in long-term human pancreatic nestin-positive cell cultures
author Maria-Engler, Sylvia Stucchi
author_facet Maria-Engler, Sylvia Stucchi
Correa-Giannella, Maria Lucia C
Labriola, Leticia
Krogh, Karin
Colin, Christian
Lojudice, Fernando Henrique
Aita, Carlos Alberto Mayora
Oliveira, Elizabeth Maria Costa de
Correa, Tatiana C Silveira
Silva, Irenice Cairo da
Genzini, Tercio
Miranda, Marcelo Perosa de
Noronha, Irene Lourdes
Vilela, Luciano
Coimbra, Cassio Negro
Mortara, Renato A. [UNIFESP]
Guia, Marcos Mares
Eliaschewitz, Freddy Goldenberg
Sogayar, Mari Cleide
author_role author
author2 Correa-Giannella, Maria Lucia C
Labriola, Leticia
Krogh, Karin
Colin, Christian
Lojudice, Fernando Henrique
Aita, Carlos Alberto Mayora
Oliveira, Elizabeth Maria Costa de
Correa, Tatiana C Silveira
Silva, Irenice Cairo da
Genzini, Tercio
Miranda, Marcelo Perosa de
Noronha, Irene Lourdes
Vilela, Luciano
Coimbra, Cassio Negro
Mortara, Renato A. [UNIFESP]
Guia, Marcos Mares
Eliaschewitz, Freddy Goldenberg
Sogayar, Mari Cleide
author2_role author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
dc.contributor.none.fl_str_mv Universidade de São Paulo (USP)
Albert Einstein Hosp
Biomm SA
Universidade Federal de São Paulo (UNIFESP)
dc.contributor.author.fl_str_mv Maria-Engler, Sylvia Stucchi
Correa-Giannella, Maria Lucia C
Labriola, Leticia
Krogh, Karin
Colin, Christian
Lojudice, Fernando Henrique
Aita, Carlos Alberto Mayora
Oliveira, Elizabeth Maria Costa de
Correa, Tatiana C Silveira
Silva, Irenice Cairo da
Genzini, Tercio
Miranda, Marcelo Perosa de
Noronha, Irene Lourdes
Vilela, Luciano
Coimbra, Cassio Negro
Mortara, Renato A. [UNIFESP]
Guia, Marcos Mares
Eliaschewitz, Freddy Goldenberg
Sogayar, Mari Cleide
description Strategies to differentiate progenitor cells into beta cells in vitro have been considered as an alternative to increase beta cell availability prior to transplantation. It has recently been suggested that nestin-positive cells could be multipotential stern cells capable of expressing endocrine markers upon specific stimulation; however, this issue still remains controversial. Here, we characterized short- and long-term islet cell cultures derived from three different human islet preparations. with respect to expression of nestin and islet cell markers, using confocal microscopy and semi-quantitative PT-PCR. the number of nestin-positive cells was found to be strikingly high in long-term cultures. in addition, a large proportion (49.7%) of these nestin-positive cells, present in long-term culture, are shown to be proliferative, as judged by BrdU incorporation. the proportion of insulin-positive cells was found to be high in short-term (up to 28 days) cultures and declined thereafter, when cells were maintained in the presence of 10% serum, concomitantly with the decrease in insulin and PDX-1 expression. Interestingly, insulin and nestin co-expression was observed as a rare event in a small proportion of cells present in freshly isolated human islets as well as in purified islet cells cultured in vitro for long periods of time. in addition, upon long-term subculturing of nestin-positive cells in 10% serum, we observed reappearance of insulin expression at the mRNA level; when these cultures were shifted to 1% serum for a month, expression of insulin, glucagon and somatostatin was also detected, indicating that manipulating the culture conditions can be used to modulate the nestin-positive cell's fate. Attempts to induce cell differentiation by plating nestin-positive cells onto Matrigel revealed that these cells tend to aggregate to form islet-like clusters, but this is not sufficient to increase insulin expression upon short-term culture. Our data corroborate previous findings indicating that, at least ill vitro, nestin-positive cells may undergo the early stages of differentiation to an islet cell phenotype and that long-term cultures of nestin-positive human islet cells may be considered as a potential source of precursor cells to generate fully differentiated/functional beta cells.
publishDate 2004
dc.date.none.fl_str_mv 2004-12-01
2016-01-24T12:37:30Z
2016-01-24T12:37:30Z
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.uri.fl_str_mv http://dx.doi.org/10.1677/joe.1.05703
Journal of Endocrinology. Bristol: Soc Endocrinology, v. 183, n. 3, p. 455-467, 2004.
10.1677/joe.1.05703
0022-0795
http://repositorio.unifesp.br/handle/11600/28032
WOS:000225947100003
url http://dx.doi.org/10.1677/joe.1.05703
http://repositorio.unifesp.br/handle/11600/28032
identifier_str_mv Journal of Endocrinology. Bristol: Soc Endocrinology, v. 183, n. 3, p. 455-467, 2004.
10.1677/joe.1.05703
0022-0795
WOS:000225947100003
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv Journal of Endocrinology
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv 455-467
dc.publisher.none.fl_str_mv Soc Endocrinology
publisher.none.fl_str_mv Soc Endocrinology
dc.source.none.fl_str_mv reponame:Repositório Institucional da UNIFESP
instname:Universidade Federal de São Paulo (UNIFESP)
instacron:UNIFESP
instname_str Universidade Federal de São Paulo (UNIFESP)
instacron_str UNIFESP
institution UNIFESP
reponame_str Repositório Institucional da UNIFESP
collection Repositório Institucional da UNIFESP
repository.name.fl_str_mv Repositório Institucional da UNIFESP - Universidade Federal de São Paulo (UNIFESP)
repository.mail.fl_str_mv biblioteca.csp@unifesp.br
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