A new experimental protocol for preferential differentiation of mouse embryonic stem cells into insulin-producing cells

Autores
Naojuk, Ortwin; Francini, Flavio; Picton,Sally; Jörns, Anne; Bailey, Clifford J.; Lenzen, Sigurd
Año de publicación
2008
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Mouse embiyonic stem (ES) cells have the potential to differentiate into insulin-producing cells, but efficient protocols for in vitro differentiation have not been established. Here we have developed a new optimized four-stage differentiation protocol and compared this with an established reference protocol. The new protocol minimized differentiation towards neuronal progeny, resulting in a population of insulin-producing cells with ß-cell characteristics but lacking neuronal features. The yield of glucagon and somatostatin cells was negligible. Crucial for this improved yield was the removal of a nestin selection step as well as removal of culture supplements that promote differentiation towards the neuronal lineage. Supplementation of the differentiation medium with insulin and fetal calf serum was beneficial for differentiation towards monohor-monal insulin-positive cells. After implantation into diabetic mice these insulin-producing cells produced a time-dependent improvement of the diabetic metabolic state, in contrast to cells differentiated according to the reference protocol. Using a spinner culture instead of an adherent culture of ES cells prevented the differentiation towards insulin-producing cells. Thus, prevention of cell attachment in a spinner culture represents a means to keep ES cells in an undifferentiated state and to inhibit differentiation. In conclusion, this study describes a new optimized four-stage protocol for differentiating ES cells to insulin-producing cells with minimal neuronal cell formation.
Fil: Naojuk, Ortwin. Medizinische Hochschule Hannover; Alemania
Fil: Francini, Flavio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Endocrinología Experimental y Aplicada. Universidad Nacional de La Plata. Facultad de Ciencias Médicas. Centro de Endocrinología Experimental y Aplicada; Argentina. Medizinische Hochschule Hannover; Alemania
Fil: Picton,Sally. Aston University. School of Life and Health Sciences; Reino Unido
Fil: Jörns, Anne. Medizinische Hochschule Hannover; Alemania
Fil: Bailey, Clifford J.. Aston University. School of Life and Health Sciences; Reino Unido
Fil: Lenzen, Sigurd. Medizinische Hochschule Hannover; Alemania
Materia
DIABETES
DIFFERENTIATION
EMBRYONIC STEM CELLS
INSULIN
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:11336/129608
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/129608
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling A new experimental protocol for preferential differentiation of mouse embryonic stem cells into insulin-producing cellsNaojuk, OrtwinFrancini, FlavioPicton,SallyJörns, AnneBailey, Clifford J.Lenzen, SigurdDIABETESDIFFERENTIATIONEMBRYONIC STEM CELLSINSULINhttps://purl.org/becyt/ford/3.2https://purl.org/becyt/ford/3Mouse embiyonic stem (ES) cells have the potential to differentiate into insulin-producing cells, but efficient protocols for in vitro differentiation have not been established. Here we have developed a new optimized four-stage differentiation protocol and compared this with an established reference protocol. The new protocol minimized differentiation towards neuronal progeny, resulting in a population of insulin-producing cells with ß-cell characteristics but lacking neuronal features. The yield of glucagon and somatostatin cells was negligible. Crucial for this improved yield was the removal of a nestin selection step as well as removal of culture supplements that promote differentiation towards the neuronal lineage. Supplementation of the differentiation medium with insulin and fetal calf serum was beneficial for differentiation towards monohor-monal insulin-positive cells. After implantation into diabetic mice these insulin-producing cells produced a time-dependent improvement of the diabetic metabolic state, in contrast to cells differentiated according to the reference protocol. Using a spinner culture instead of an adherent culture of ES cells prevented the differentiation towards insulin-producing cells. Thus, prevention of cell attachment in a spinner culture represents a means to keep ES cells in an undifferentiated state and to inhibit differentiation. In conclusion, this study describes a new optimized four-stage protocol for differentiating ES cells to insulin-producing cells with minimal neuronal cell formation.Fil: Naojuk, Ortwin. Medizinische Hochschule Hannover; AlemaniaFil: Francini, Flavio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Endocrinología Experimental y Aplicada. Universidad Nacional de La Plata. Facultad de Ciencias Médicas. Centro de Endocrinología Experimental y Aplicada; Argentina. Medizinische Hochschule Hannover; AlemaniaFil: Picton,Sally. Aston University. School of Life and Health Sciences; Reino UnidoFil: Jörns, Anne. Medizinische Hochschule Hannover; AlemaniaFil: Bailey, Clifford J.. Aston University. School of Life and Health Sciences; Reino UnidoFil: Lenzen, Sigurd. Medizinische Hochschule Hannover; AlemaniaCognizant Communication Corp2008-12info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/129608Naojuk, Ortwin; Francini, Flavio; Picton,Sally; Jörns, Anne; Bailey, Clifford J.; et al.; A new experimental protocol for preferential differentiation of mouse embryonic stem cells into insulin-producing cells; Cognizant Communication Corp; Cell Transplantation; 17; 10-11; 12-2008; 1231-12420963-6897CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/doi/10.3727/096368908787236549info:eu-repo/semantics/altIdentifier/url/https://journals.sagepub.com/doi/10.3727/096368908787236549info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-03T09:57:11Zoai:ri.conicet.gov.ar:11336/129608instacron: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:34982025-09-03 09:57:12.029CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv A new experimental protocol for preferential differentiation of mouse embryonic stem cells into insulin-producing cells
title A new experimental protocol for preferential differentiation of mouse embryonic stem cells into insulin-producing cells
spellingShingle A new experimental protocol for preferential differentiation of mouse embryonic stem cells into insulin-producing cells
Naojuk, Ortwin
DIABETES
DIFFERENTIATION
EMBRYONIC STEM CELLS
INSULIN
title_short A new experimental protocol for preferential differentiation of mouse embryonic stem cells into insulin-producing cells
title_full A new experimental protocol for preferential differentiation of mouse embryonic stem cells into insulin-producing cells
title_fullStr A new experimental protocol for preferential differentiation of mouse embryonic stem cells into insulin-producing cells
title_full_unstemmed A new experimental protocol for preferential differentiation of mouse embryonic stem cells into insulin-producing cells
title_sort A new experimental protocol for preferential differentiation of mouse embryonic stem cells into insulin-producing cells
dc.creator.none.fl_str_mv Naojuk, Ortwin
Francini, Flavio
Picton,Sally
Jörns, Anne
Bailey, Clifford J.
Lenzen, Sigurd
author Naojuk, Ortwin
author_facet Naojuk, Ortwin
Francini, Flavio
Picton,Sally
Jörns, Anne
Bailey, Clifford J.
Lenzen, Sigurd
author_role author
author2 Francini, Flavio
Picton,Sally
Jörns, Anne
Bailey, Clifford J.
Lenzen, Sigurd
author2_role author
author
author
author
author
dc.subject.none.fl_str_mv DIABETES
DIFFERENTIATION
EMBRYONIC STEM CELLS
INSULIN
topic DIABETES
DIFFERENTIATION
EMBRYONIC STEM CELLS
INSULIN
purl_subject.fl_str_mv https://purl.org/becyt/ford/3.2
https://purl.org/becyt/ford/3
dc.description.none.fl_txt_mv Mouse embiyonic stem (ES) cells have the potential to differentiate into insulin-producing cells, but efficient protocols for in vitro differentiation have not been established. Here we have developed a new optimized four-stage differentiation protocol and compared this with an established reference protocol. The new protocol minimized differentiation towards neuronal progeny, resulting in a population of insulin-producing cells with ß-cell characteristics but lacking neuronal features. The yield of glucagon and somatostatin cells was negligible. Crucial for this improved yield was the removal of a nestin selection step as well as removal of culture supplements that promote differentiation towards the neuronal lineage. Supplementation of the differentiation medium with insulin and fetal calf serum was beneficial for differentiation towards monohor-monal insulin-positive cells. After implantation into diabetic mice these insulin-producing cells produced a time-dependent improvement of the diabetic metabolic state, in contrast to cells differentiated according to the reference protocol. Using a spinner culture instead of an adherent culture of ES cells prevented the differentiation towards insulin-producing cells. Thus, prevention of cell attachment in a spinner culture represents a means to keep ES cells in an undifferentiated state and to inhibit differentiation. In conclusion, this study describes a new optimized four-stage protocol for differentiating ES cells to insulin-producing cells with minimal neuronal cell formation.
Fil: Naojuk, Ortwin. Medizinische Hochschule Hannover; Alemania
Fil: Francini, Flavio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Endocrinología Experimental y Aplicada. Universidad Nacional de La Plata. Facultad de Ciencias Médicas. Centro de Endocrinología Experimental y Aplicada; Argentina. Medizinische Hochschule Hannover; Alemania
Fil: Picton,Sally. Aston University. School of Life and Health Sciences; Reino Unido
Fil: Jörns, Anne. Medizinische Hochschule Hannover; Alemania
Fil: Bailey, Clifford J.. Aston University. School of Life and Health Sciences; Reino Unido
Fil: Lenzen, Sigurd. Medizinische Hochschule Hannover; Alemania
description Mouse embiyonic stem (ES) cells have the potential to differentiate into insulin-producing cells, but efficient protocols for in vitro differentiation have not been established. Here we have developed a new optimized four-stage differentiation protocol and compared this with an established reference protocol. The new protocol minimized differentiation towards neuronal progeny, resulting in a population of insulin-producing cells with ß-cell characteristics but lacking neuronal features. The yield of glucagon and somatostatin cells was negligible. Crucial for this improved yield was the removal of a nestin selection step as well as removal of culture supplements that promote differentiation towards the neuronal lineage. Supplementation of the differentiation medium with insulin and fetal calf serum was beneficial for differentiation towards monohor-monal insulin-positive cells. After implantation into diabetic mice these insulin-producing cells produced a time-dependent improvement of the diabetic metabolic state, in contrast to cells differentiated according to the reference protocol. Using a spinner culture instead of an adherent culture of ES cells prevented the differentiation towards insulin-producing cells. Thus, prevention of cell attachment in a spinner culture represents a means to keep ES cells in an undifferentiated state and to inhibit differentiation. In conclusion, this study describes a new optimized four-stage protocol for differentiating ES cells to insulin-producing cells with minimal neuronal cell formation.
publishDate 2008
dc.date.none.fl_str_mv 2008-12
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/11336/129608
Naojuk, Ortwin; Francini, Flavio; Picton,Sally; Jörns, Anne; Bailey, Clifford J.; et al.; A new experimental protocol for preferential differentiation of mouse embryonic stem cells into insulin-producing cells; Cognizant Communication Corp; Cell Transplantation; 17; 10-11; 12-2008; 1231-1242
0963-6897
CONICET Digital
CONICET
url http://hdl.handle.net/11336/129608
identifier_str_mv Naojuk, Ortwin; Francini, Flavio; Picton,Sally; Jörns, Anne; Bailey, Clifford J.; et al.; A new experimental protocol for preferential differentiation of mouse embryonic stem cells into insulin-producing cells; Cognizant Communication Corp; Cell Transplantation; 17; 10-11; 12-2008; 1231-1242
0963-6897
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/doi/10.3727/096368908787236549
info:eu-repo/semantics/altIdentifier/url/https://journals.sagepub.com/doi/10.3727/096368908787236549
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
dc.publisher.none.fl_str_mv Cognizant Communication Corp
publisher.none.fl_str_mv Cognizant Communication Corp
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
_version_ 1842269446892158976
score 13.13397

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