Selective effect of INGAP-PP upon mouse embryonic stem cell differentiation toward islet cells

Autores
Francini, Flavio; del Zotto, Hector Herminio; Massa, Maria Laura; Gagliardino, Juan Jose
Año de publicación
2009
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
We evaluated the effect of islet neogenesis-associated protein pentadecapeptide (INGAP-PP) upon islet β- and non-β cell differentiation from mouse embryonic stem (mES) cells. ES-D3 cell lines were cultured following Lumelsky's protocol with or without INGAP-PP (5 µg/ml) at different stages. Gene expression was quantified using qPCR. mES cells were fixed and immunostained using anti insulin-, somatostatin-, glucagon-, Pdx-1-, Ngn-3-, Nkx-6.1 and PGP9.5 specific antibodies. PCNA was used to measure replication rate. Bcl2 (immunostaining) and caspase-3 (enzyme activity and gene expression) were determined as apoptosis markers. INGAP-PP increased IAPP, Glut-2, Kir-6.2, SUR-1 and insulin gene expression, and the percentage of insulin-immunostained cells. Conversely, INGAP-PP reduced significantly glucagon and somatostatin gene expression and immunopositivity. While nestin gene expression was not affected, there was a significant reduction in the percentage of PGP9.5-immunostained cells. Pdx-1 gene expression increased by 115% in INGAP-PP treated cells, as well as the percentage of Pdx-1, Ngn-3 and Nkx-6.1 immunopositive cells. Neither caspase-3 (expression and activity) nor Bcl2 positively immunostained cells were affected by INGAP-PP. Accordingly, INGAP-PP would promote stem cell differentiation into a β-like cell phenotype, simultaneously decreasing its differentiation toward non-β-cell precursors. Therefore, INGAP-PP would be potentially useful to obtain β-cells from stem cells for replacement therapy.
Fil: Francini, Flavio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico La Plata. Centro de Endocrinología Experimental y Aplicada (i); Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Médicas; Argentina
Fil: del Zotto, Hector Herminio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico La Plata. Centro de Endocrinología Experimental y Aplicada (i); Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Médicas; Argentina
Fil: Massa, Maria Laura. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico La Plata. Centro de Endocrinología Experimental y Aplicada (i); Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Médicas; Argentina
Fil: Gagliardino, Juan Jose. Universidad Nacional de La Plata. Facultad de Ciencias Médicas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico La Plata. Centro de Endocrinología Experimental y Aplicada (i); Argentina
Materia
Embryonic stem cells
Insulin-producing cells
Differentiation
INGAP-PP
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/95548
Acceder
id CONICETDig_8fca0fec9246d0fdfd848220eb7b1ca8
oai_identifier_str oai:ri.conicet.gov.ar:11336/95548
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Selective effect of INGAP-PP upon mouse embryonic stem cell differentiation toward islet cellsFrancini, Flaviodel Zotto, Hector HerminioMassa, Maria LauraGagliardino, Juan JoseEmbryonic stem cellsInsulin-producing cellsDifferentiationINGAP-PPhttps://purl.org/becyt/ford/3.1https://purl.org/becyt/ford/3https://purl.org/becyt/ford/3.1https://purl.org/becyt/ford/3We evaluated the effect of islet neogenesis-associated protein pentadecapeptide (INGAP-PP) upon islet β- and non-β cell differentiation from mouse embryonic stem (mES) cells. ES-D3 cell lines were cultured following Lumelsky's protocol with or without INGAP-PP (5 µg/ml) at different stages. Gene expression was quantified using qPCR. mES cells were fixed and immunostained using anti insulin-, somatostatin-, glucagon-, Pdx-1-, Ngn-3-, Nkx-6.1 and PGP9.5 specific antibodies. PCNA was used to measure replication rate. Bcl2 (immunostaining) and caspase-3 (enzyme activity and gene expression) were determined as apoptosis markers. INGAP-PP increased IAPP, Glut-2, Kir-6.2, SUR-1 and insulin gene expression, and the percentage of insulin-immunostained cells. Conversely, INGAP-PP reduced significantly glucagon and somatostatin gene expression and immunopositivity. While nestin gene expression was not affected, there was a significant reduction in the percentage of PGP9.5-immunostained cells. Pdx-1 gene expression increased by 115% in INGAP-PP treated cells, as well as the percentage of Pdx-1, Ngn-3 and Nkx-6.1 immunopositive cells. Neither caspase-3 (expression and activity) nor Bcl2 positively immunostained cells were affected by INGAP-PP. Accordingly, INGAP-PP would promote stem cell differentiation into a β-like cell phenotype, simultaneously decreasing its differentiation toward non-β-cell precursors. Therefore, INGAP-PP would be potentially useful to obtain β-cells from stem cells for replacement therapy.Fil: Francini, Flavio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico La Plata. Centro de Endocrinología Experimental y Aplicada (i); Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Médicas; ArgentinaFil: del Zotto, Hector Herminio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico La Plata. Centro de Endocrinología Experimental y Aplicada (i); Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Médicas; ArgentinaFil: Massa, Maria Laura. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico La Plata. Centro de Endocrinología Experimental y Aplicada (i); Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Médicas; ArgentinaFil: Gagliardino, Juan Jose. Universidad Nacional de La Plata. Facultad de Ciencias Médicas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico La Plata. Centro de Endocrinología Experimental y Aplicada (i); ArgentinaElsevier Science2009-02info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/95548Francini, Flavio; del Zotto, Hector Herminio; Massa, Maria Laura; Gagliardino, Juan Jose; Selective effect of INGAP-PP upon mouse embryonic stem cell differentiation toward islet cells; Elsevier Science; Regulatory Peptides; 153; 1-3; 2-2009; 43-480167-0115CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://www.sciencedirect.com/science/article/pii/S016701150800219Xinfo:eu-repo/semantics/altIdentifier/doi/10.1016/j.regpep.2008.12.006info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T10:45:42Zoai:ri.conicet.gov.ar:11336/95548instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-09-29 10:45:42.635CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Selective effect of INGAP-PP upon mouse embryonic stem cell differentiation toward islet cells
title Selective effect of INGAP-PP upon mouse embryonic stem cell differentiation toward islet cells
spellingShingle Selective effect of INGAP-PP upon mouse embryonic stem cell differentiation toward islet cells
Francini, Flavio
Embryonic stem cells
Insulin-producing cells
Differentiation
INGAP-PP
title_short Selective effect of INGAP-PP upon mouse embryonic stem cell differentiation toward islet cells
title_full Selective effect of INGAP-PP upon mouse embryonic stem cell differentiation toward islet cells
title_fullStr Selective effect of INGAP-PP upon mouse embryonic stem cell differentiation toward islet cells
title_full_unstemmed Selective effect of INGAP-PP upon mouse embryonic stem cell differentiation toward islet cells
title_sort Selective effect of INGAP-PP upon mouse embryonic stem cell differentiation toward islet cells
dc.creator.none.fl_str_mv Francini, Flavio
del Zotto, Hector Herminio
Massa, Maria Laura
Gagliardino, Juan Jose
author Francini, Flavio
author_facet Francini, Flavio
del Zotto, Hector Herminio
Massa, Maria Laura
Gagliardino, Juan Jose
author_role author
author2 del Zotto, Hector Herminio
Massa, Maria Laura
Gagliardino, Juan Jose
author2_role author
author
author
dc.subject.none.fl_str_mv Embryonic stem cells
Insulin-producing cells
Differentiation
INGAP-PP
topic Embryonic stem cells
Insulin-producing cells
Differentiation
INGAP-PP
purl_subject.fl_str_mv https://purl.org/becyt/ford/3.1
https://purl.org/becyt/ford/3
https://purl.org/becyt/ford/3.1
https://purl.org/becyt/ford/3
dc.description.none.fl_txt_mv We evaluated the effect of islet neogenesis-associated protein pentadecapeptide (INGAP-PP) upon islet β- and non-β cell differentiation from mouse embryonic stem (mES) cells. ES-D3 cell lines were cultured following Lumelsky's protocol with or without INGAP-PP (5 µg/ml) at different stages. Gene expression was quantified using qPCR. mES cells were fixed and immunostained using anti insulin-, somatostatin-, glucagon-, Pdx-1-, Ngn-3-, Nkx-6.1 and PGP9.5 specific antibodies. PCNA was used to measure replication rate. Bcl2 (immunostaining) and caspase-3 (enzyme activity and gene expression) were determined as apoptosis markers. INGAP-PP increased IAPP, Glut-2, Kir-6.2, SUR-1 and insulin gene expression, and the percentage of insulin-immunostained cells. Conversely, INGAP-PP reduced significantly glucagon and somatostatin gene expression and immunopositivity. While nestin gene expression was not affected, there was a significant reduction in the percentage of PGP9.5-immunostained cells. Pdx-1 gene expression increased by 115% in INGAP-PP treated cells, as well as the percentage of Pdx-1, Ngn-3 and Nkx-6.1 immunopositive cells. Neither caspase-3 (expression and activity) nor Bcl2 positively immunostained cells were affected by INGAP-PP. Accordingly, INGAP-PP would promote stem cell differentiation into a β-like cell phenotype, simultaneously decreasing its differentiation toward non-β-cell precursors. Therefore, INGAP-PP would be potentially useful to obtain β-cells from stem cells for replacement therapy.
Fil: Francini, Flavio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico La Plata. Centro de Endocrinología Experimental y Aplicada (i); Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Médicas; Argentina
Fil: del Zotto, Hector Herminio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico La Plata. Centro de Endocrinología Experimental y Aplicada (i); Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Médicas; Argentina
Fil: Massa, Maria Laura. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico La Plata. Centro de Endocrinología Experimental y Aplicada (i); Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Médicas; Argentina
Fil: Gagliardino, Juan Jose. Universidad Nacional de La Plata. Facultad de Ciencias Médicas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico La Plata. Centro de Endocrinología Experimental y Aplicada (i); Argentina
description We evaluated the effect of islet neogenesis-associated protein pentadecapeptide (INGAP-PP) upon islet β- and non-β cell differentiation from mouse embryonic stem (mES) cells. ES-D3 cell lines were cultured following Lumelsky's protocol with or without INGAP-PP (5 µg/ml) at different stages. Gene expression was quantified using qPCR. mES cells were fixed and immunostained using anti insulin-, somatostatin-, glucagon-, Pdx-1-, Ngn-3-, Nkx-6.1 and PGP9.5 specific antibodies. PCNA was used to measure replication rate. Bcl2 (immunostaining) and caspase-3 (enzyme activity and gene expression) were determined as apoptosis markers. INGAP-PP increased IAPP, Glut-2, Kir-6.2, SUR-1 and insulin gene expression, and the percentage of insulin-immunostained cells. Conversely, INGAP-PP reduced significantly glucagon and somatostatin gene expression and immunopositivity. While nestin gene expression was not affected, there was a significant reduction in the percentage of PGP9.5-immunostained cells. Pdx-1 gene expression increased by 115% in INGAP-PP treated cells, as well as the percentage of Pdx-1, Ngn-3 and Nkx-6.1 immunopositive cells. Neither caspase-3 (expression and activity) nor Bcl2 positively immunostained cells were affected by INGAP-PP. Accordingly, INGAP-PP would promote stem cell differentiation into a β-like cell phenotype, simultaneously decreasing its differentiation toward non-β-cell precursors. Therefore, INGAP-PP would be potentially useful to obtain β-cells from stem cells for replacement therapy.
publishDate 2009
dc.date.none.fl_str_mv 2009-02
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
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://hdl.handle.net/11336/95548
Francini, Flavio; del Zotto, Hector Herminio; Massa, Maria Laura; Gagliardino, Juan Jose; Selective effect of INGAP-PP upon mouse embryonic stem cell differentiation toward islet cells; Elsevier Science; Regulatory Peptides; 153; 1-3; 2-2009; 43-48
0167-0115
CONICET Digital
CONICET
url http://hdl.handle.net/11336/95548
identifier_str_mv Francini, Flavio; del Zotto, Hector Herminio; Massa, Maria Laura; Gagliardino, Juan Jose; Selective effect of INGAP-PP upon mouse embryonic stem cell differentiation toward islet cells; Elsevier Science; Regulatory Peptides; 153; 1-3; 2-2009; 43-48
0167-0115
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/https://www.sciencedirect.com/science/article/pii/S016701150800219X
info:eu-repo/semantics/altIdentifier/doi/10.1016/j.regpep.2008.12.006
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
application/pdf
application/pdf
application/pdf
dc.publisher.none.fl_str_mv Elsevier Science
publisher.none.fl_str_mv Elsevier Science
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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score 13.070432

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