Details

Autor(en) / Beteiligte

• Maria-Engler, Silvya Stuchi
• Corrêa-Giannella, Maria Lúcia C
• Labriola, Letícia
• Krogh, Karin
• Colin, Christian
• Lojudice, Fernando Henrique
• Aita, Carlos Alberto Mayora
• de Oliveira, Elizabeth Maria Costa
• Corrêa, Tatiana C Silveira
• da Silva, Irenice Cairo
• Genzini, Tercio
• de Miranda, Marcelo Perosa
• Noronha, Irene Lourdes
• Vilela, Luciano
• Coimbra, Cassio Negro
• Mortara, Renato A
• Mares Guia, Marcos
• Eliaschewitz, Freddy Goldberg
• Sogayar, Mari Cleide

Titel

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

Ist Teil von

• Journal of endocrinology, 2004-12, Vol.183 (3), p.455-467

Ort / Verlag

Colchester: BioScientifica

Erscheinungsjahr

2004

Quelle

MEDLINE

Beschreibungen/Notizen

• Strategies to differentiate progenitor cells into β cells in vitro have been considered as an alternative to increase β cell availability prior to transplantation. It has recently been suggested that nestin-positive cells could be multipotential stem 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 RT-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 in 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 β cells.