Obtaining hepatocyte-like cells from dental pulp pluripotent-like stem cells

The liver is the largest internal organ and an important regulator of homeostasis providing essential metabolic, exocrine and endocrine functions. Liver diseases result in high rates of morbidity and mortality and affect a growing number of people over the years. Obtaining mature hepatocytes from st...

Descripción completa

Detalles Bibliográficos
Autor: Gil Recio, Carlos
Tipo de recurso: tesis doctoral
Estado:Versión publicada
Fecha de publicación:2015
País:España
Institución:CBUC, CESCA
Repositorio:TDR. Tesis Doctorales en Red
OAI Identifier:oai:www.tdx.cat:10803/299800
Acceso en línea:http://hdl.handle.net/10803/299800
Access Level:acceso abierto
Palabra clave:Stem cells
Hepatocytes
Dental pulp
Liver
Pluripotent
Medicina Regenerativa
579
61
id ES_30e7a2bb2e7e505651bf94e07439bf70
oai_identifier_str oai:www.tdx.cat:10803/299800
network_acronym_str ES
network_name_str España
repository_id_str
spelling Obtaining hepatocyte-like cells from dental pulp pluripotent-like stem cellsGil Recio, CarlosStem cellsHepatocytesDental pulpLiverPluripotentMedicina Regenerativa57961The liver is the largest internal organ and an important regulator of homeostasis providing essential metabolic, exocrine and endocrine functions. Liver diseases result in high rates of morbidity and mortality and affect a growing number of people over the years. Obtaining mature hepatocytes from stem cells is a major goal in regenerative medicine studies due to the clinical applications that could derive from it. Dental pulp pluripotent stem cells are a considerable promise in tissue engineering and regenerative medicine as a source of tissue-specific cells, therefore the aim of this study is to demonstrate their ability to generate functional hepatocyte-like cells in vitro. Cells were cultured in a 2D environment and they were supplemented with several growth factors that recapitulate liver development. The expression of hepatic markers was detected by qRT-PCR and was confirmed by immunofluorescence analysis. The functional efficacy of the differentiated hepatic cells was evaluated by the secretion levels of albumin, cytochrome P450 activity, glycogen storage capacity and the production of hepatic enzymes was also evaluated. In this study we developed a 2D differentiation protocol of DPPSCs into hepatic lineage in vitro. The protocol was optimized to obtain definitive endoderm during the first days of differentiation and was continued with a modified five-step induction medium for 22 days based on liver development. Differentiated cells were able to express hepatic proteins during the entire differentiation process and were able to carry out several functions expected in mature hepatocytes. In this study we demonstrate that DPPSCs can be a powerful tool for liver regenerative medicine treatments to be developed in the future.Universitat Internacional de CatalunyaAtari Abouassi, MaherBarajas Vélez, MiguelMontori Pina, SheylaUniversitat Internacional de Catalunya. Departament de Ciències Bàsiques201520152015info:eu-repo/semantics/doctoralThesisinfo:eu-repo/semantics/publishedVersion150 p.application/pdfapplication/pdfhttp://hdl.handle.net/10803/299800TDX (Tesis Doctorals en Xarxa)reponame:TDR. Tesis Doctorales en Redinstname:CBUC, CESCAInglésL'accés als continguts d'aquesta tesi queda condicionat a l'acceptació de les condicions d'ús establertes per la següent llicència Creative Commons: http://creativecommons.org/licenses/by-nc-nd/3.0/es/http://creativecommons.org/licenses/by-nc-nd/3.0/es/info:eu-repo/semantics/openAccessoai:www.tdx.cat:10803/2998002026-06-14T12:46:07Z
dc.title.none.fl_str_mv Obtaining hepatocyte-like cells from dental pulp pluripotent-like stem cells
title Obtaining hepatocyte-like cells from dental pulp pluripotent-like stem cells
spellingShingle Obtaining hepatocyte-like cells from dental pulp pluripotent-like stem cells
Gil Recio, Carlos
Stem cells
Hepatocytes
Dental pulp
Liver
Pluripotent
Medicina Regenerativa
579
61
title_short Obtaining hepatocyte-like cells from dental pulp pluripotent-like stem cells
title_full Obtaining hepatocyte-like cells from dental pulp pluripotent-like stem cells
title_fullStr Obtaining hepatocyte-like cells from dental pulp pluripotent-like stem cells
title_full_unstemmed Obtaining hepatocyte-like cells from dental pulp pluripotent-like stem cells
title_sort Obtaining hepatocyte-like cells from dental pulp pluripotent-like stem cells
dc.creator.none.fl_str_mv Gil Recio, Carlos
author Gil Recio, Carlos
author_facet Gil Recio, Carlos
author_role author
dc.contributor.none.fl_str_mv Atari Abouassi, Maher
Barajas Vélez, Miguel
Montori Pina, Sheyla
Universitat Internacional de Catalunya. Departament de Ciències Bàsiques
dc.subject.none.fl_str_mv Stem cells
Hepatocytes
Dental pulp
Liver
Pluripotent
Medicina Regenerativa
579
61
topic Stem cells
Hepatocytes
Dental pulp
Liver
Pluripotent
Medicina Regenerativa
579
61
description The liver is the largest internal organ and an important regulator of homeostasis providing essential metabolic, exocrine and endocrine functions. Liver diseases result in high rates of morbidity and mortality and affect a growing number of people over the years. Obtaining mature hepatocytes from stem cells is a major goal in regenerative medicine studies due to the clinical applications that could derive from it. Dental pulp pluripotent stem cells are a considerable promise in tissue engineering and regenerative medicine as a source of tissue-specific cells, therefore the aim of this study is to demonstrate their ability to generate functional hepatocyte-like cells in vitro. Cells were cultured in a 2D environment and they were supplemented with several growth factors that recapitulate liver development. The expression of hepatic markers was detected by qRT-PCR and was confirmed by immunofluorescence analysis. The functional efficacy of the differentiated hepatic cells was evaluated by the secretion levels of albumin, cytochrome P450 activity, glycogen storage capacity and the production of hepatic enzymes was also evaluated. In this study we developed a 2D differentiation protocol of DPPSCs into hepatic lineage in vitro. The protocol was optimized to obtain definitive endoderm during the first days of differentiation and was continued with a modified five-step induction medium for 22 days based on liver development. Differentiated cells were able to express hepatic proteins during the entire differentiation process and were able to carry out several functions expected in mature hepatocytes. In this study we demonstrate that DPPSCs can be a powerful tool for liver regenerative medicine treatments to be developed in the future.
publishDate 2015
dc.date.none.fl_str_mv 2015
2015
2015
dc.type.none.fl_str_mv info:eu-repo/semantics/doctoralThesis
info:eu-repo/semantics/publishedVersion
format doctoralThesis
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/10803/299800
url http://hdl.handle.net/10803/299800
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.rights.none.fl_str_mv http://creativecommons.org/licenses/by-nc-nd/3.0/es/
info:eu-repo/semantics/openAccess
rights_invalid_str_mv http://creativecommons.org/licenses/by-nc-nd/3.0/es/
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv 150 p.
application/pdf
application/pdf
dc.publisher.none.fl_str_mv Universitat Internacional de Catalunya
publisher.none.fl_str_mv Universitat Internacional de Catalunya
dc.source.none.fl_str_mv TDX (Tesis Doctorals en Xarxa)
reponame:TDR. Tesis Doctorales en Red
instname:CBUC, CESCA
instname_str CBUC, CESCA
reponame_str TDR. Tesis Doctorales en Red
collection TDR. Tesis Doctorales en Red
repository.name.fl_str_mv
repository.mail.fl_str_mv
_version_ 1869405575746420736
score 15,300719

Similares en LA Referencia