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

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

Autor Maria-Engler, Sylvia Stucchi Google Scholar
Correa-Giannella, Maria Lucia C Google Scholar
Labriola, Leticia Google Scholar
Krogh, Karin Google Scholar
Colin, Christian Google Scholar
Lojudice, Fernando Henrique Google Scholar
Aita, Carlos Alberto Mayora Google Scholar
Oliveira, Elizabeth Maria Costa de Google Scholar
Correa, Tatiana C Silveira Google Scholar
Silva, Irenice Cairo da Google Scholar
Genzini, Tercio Google Scholar
Miranda, Marcelo Perosa de Google Scholar
Noronha, Irene Lourdes Google Scholar
Vilela, Luciano Google Scholar
Coimbra, Cassio Negro Google Scholar
Mortara, Renato A. Autor UNIFESP Google Scholar
Guia, Marcos Mares Google Scholar
Eliaschewitz, Freddy Goldenberg Google Scholar
Sogayar, Mari Cleide Google Scholar
Instituição Universidade de São Paulo (USP)
Albert Einstein Hosp
Biomm SA
Universidade Federal de São Paulo (UNIFESP)
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.
Idioma Inglês
Data de publicação 2004-12-01
Publicado em Journal of Endocrinology. Bristol: Soc Endocrinology, v. 183, n. 3, p. 455-467, 2004.
ISSN 0022-0795 (Sherpa/Romeo, fator de impacto)
Publicador Soc Endocrinology
Extensão 455-467
Fonte http://dx.doi.org/10.1677/joe.1.05703
Direito de acesso Acesso aberto Open Access
Tipo Artigo
Web of Science WOS:000225947100003
Endereço permanente http://repositorio.unifesp.br/handle/11600/28032

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