A deficitary model of CDK5 does not impairs neuronal differentiation of human pluripotent stem cells

Autores
Mucci, Sofia; Isaja, Luciana; Rodríguez Varela, Maria Soledad; Ferriol Laffouillere, Sofia Lujan; Sevlever, Gustavo; Scassa, Maria Elida; Romorini, Leonardo
Año de publicación
2021
Idioma
inglés
Tipo de recurso
documento de conferencia
Estado
versión publicada
Descripción
CDK5/P35 is a complex involved in neuronal homeostasis and de- velopment that was described as a critical player for neuronal surviv- al. Besides, its deregulation is linked with neurodegenerative pathol- ogies such as Alzheimer Disease and Parkinson Disease. For that reason, we generated a deficitary CDK5 genetic model in neurons derived from human pluripotent stem cells. For this purpose, we used CRISPR/Cas9 technology to generate human embryonic and induced pluripotent stem cells (hESCs and hiPSCs, respectively) KO-CDK5 lines. CDK5 protein expression levels were analyzed by western blot in samples obtained from clones where indels caused by CRISPR/Cas9 editing were detected by DNA sequencing. We obtained CDK5-/- clones for H9 hESCs and FN2.1 hiPSCs lines and a CDK5+/- clone for H9 hESCs line. Then, neural stem cells (NSC) were derived from the CDK5 KO clones using a commercial neural induction medium and their phenotype was validated by im- munofluorescence staining using antibodies that recognize specific lineage markers (SOX-1, SOX-2, NESTIN and PAX-6). Finally, NSC obtained from the heterozygous CDK5+/- KO H9 hESCs clone were differentiated into neurons using a 2D-based protocol and their phe- notype was validated by immunofluorescence staining of neuronal specific markers (TUJ-1 and MAP2). In conclusion, we managed to obtain NSC-neurons from CDK5-/- and CDK5+/- clones, determin- ing that CDK5 is not essential for NSC generation. Besides, neuro- nal differentiation was achieved for H9 CDK5+/- clone, indicating that the CDK5 deficiency does not impair the generation of NSC-derived neurons. This result allows us to account with a CDK5-defi- cient model to further study its participation in neuronal homeostasis dysfunctions.
Fil: Mucci, Sofia. Fundación para la Lucha contra las Enfermedades Neurológicas de la Infancia; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Isaja, Luciana. Fundación para la Lucha contra las Enfermedades Neurológicas de la Infancia; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Rodríguez Varela, Maria Soledad. Fundación para la Lucha contra las Enfermedades Neurológicas de la Infancia; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Ferriol Laffouillere, Sofia Lujan. Fundación para la Lucha contra las Enfermedades Neurológicas de la Infancia; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Sevlever, Gustavo. Fundación para la Lucha contra las Enfermedades Neurológicas de la Infancia; Argentina
Fil: Scassa, Maria Elida. Fundación para la Lucha contra las Enfermedades Neurológicas de la Infancia; Argentina
Fil: Romorini, Leonardo. Fundación para la Lucha contra las Enfermedades Neurológicas de la Infancia; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
LXVI Reunión Anual de la Sociedad Argentina de Investigación Clínica; LXIX Reunión Anual de la Sociedad Argentina de Inmunología; LIII Reunión Anual de la Asociación Argentina de Farmacología Experimental y XI Reunión Anual de la Asociación Argentina de Nanomedicinas
Argentina
Sociedad Argentina de Investigación Clínica
Sociedad Argentina de Inmunología
Asociación Argentina de Farmacología Experimental
Asociación Argentina de Nanomedicinas
Materia
CDK5
NEURONAL DIFFERENTIATION
HUMAN PLURIPOTENT STEM CELLS
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:11336/159144
Acceder
id CONICETDig_a5fad507a155b459553bcb6f68dad0c1
oai_identifier_str oai:ri.conicet.gov.ar:11336/159144
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling A deficitary model of CDK5 does not impairs neuronal differentiation of human pluripotent stem cellsMucci, SofiaIsaja, LucianaRodríguez Varela, Maria SoledadFerriol Laffouillere, Sofia LujanSevlever, GustavoScassa, Maria ElidaRomorini, LeonardoCDK5NEURONAL DIFFERENTIATIONHUMAN PLURIPOTENT STEM CELLShttps://purl.org/becyt/ford/3.1https://purl.org/becyt/ford/3CDK5/P35 is a complex involved in neuronal homeostasis and de- velopment that was described as a critical player for neuronal surviv- al. Besides, its deregulation is linked with neurodegenerative pathol- ogies such as Alzheimer Disease and Parkinson Disease. For that reason, we generated a deficitary CDK5 genetic model in neurons derived from human pluripotent stem cells. For this purpose, we used CRISPR/Cas9 technology to generate human embryonic and induced pluripotent stem cells (hESCs and hiPSCs, respectively) KO-CDK5 lines. CDK5 protein expression levels were analyzed by western blot in samples obtained from clones where indels caused by CRISPR/Cas9 editing were detected by DNA sequencing. We obtained CDK5-/- clones for H9 hESCs and FN2.1 hiPSCs lines and a CDK5+/- clone for H9 hESCs line. Then, neural stem cells (NSC) were derived from the CDK5 KO clones using a commercial neural induction medium and their phenotype was validated by im- munofluorescence staining using antibodies that recognize specific lineage markers (SOX-1, SOX-2, NESTIN and PAX-6). Finally, NSC obtained from the heterozygous CDK5+/- KO H9 hESCs clone were differentiated into neurons using a 2D-based protocol and their phe- notype was validated by immunofluorescence staining of neuronal specific markers (TUJ-1 and MAP2). In conclusion, we managed to obtain NSC-neurons from CDK5-/- and CDK5+/- clones, determin- ing that CDK5 is not essential for NSC generation. Besides, neuro- nal differentiation was achieved for H9 CDK5+/- clone, indicating that the CDK5 deficiency does not impair the generation of NSC-derived neurons. This result allows us to account with a CDK5-defi- cient model to further study its participation in neuronal homeostasis dysfunctions.Fil: Mucci, Sofia. Fundación para la Lucha contra las Enfermedades Neurológicas de la Infancia; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Isaja, Luciana. Fundación para la Lucha contra las Enfermedades Neurológicas de la Infancia; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Rodríguez Varela, Maria Soledad. Fundación para la Lucha contra las Enfermedades Neurológicas de la Infancia; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Ferriol Laffouillere, Sofia Lujan. Fundación para la Lucha contra las Enfermedades Neurológicas de la Infancia; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Sevlever, Gustavo. Fundación para la Lucha contra las Enfermedades Neurológicas de la Infancia; ArgentinaFil: Scassa, Maria Elida. Fundación para la Lucha contra las Enfermedades Neurológicas de la Infancia; ArgentinaFil: Romorini, Leonardo. Fundación para la Lucha contra las Enfermedades Neurológicas de la Infancia; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaLXVI Reunión Anual de la Sociedad Argentina de Investigación Clínica; LXIX Reunión Anual de la Sociedad Argentina de Inmunología; LIII Reunión Anual de la Asociación Argentina de Farmacología Experimental y XI Reunión Anual de la Asociación Argentina de NanomedicinasArgentinaSociedad Argentina de Investigación ClínicaSociedad Argentina de InmunologíaAsociación Argentina de Farmacología ExperimentalAsociación Argentina de NanomedicinasFundación Revista Medicina2021info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/conferenceObjectReuniónJournalhttp://purl.org/coar/resource_type/c_5794info:ar-repo/semantics/documentoDeConferenciaapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/159144A deficitary model of CDK5 does not impairs neuronal differentiation of human pluripotent stem cells; LXVI Reunión Anual de la Sociedad Argentina de Investigación Clínica; LXIX Reunión Anual de la Sociedad Argentina de Inmunología; LIII Reunión Anual de la Asociación Argentina de Farmacología Experimental y XI Reunión Anual de la Asociación Argentina de Nanomedicinas; Argentina; 2021; 1-5CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://medicinabuenosaires.com/revistas/vol81-21/s3/Mv81s3.pdfNacionalinfo:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T09:35:59Zoai:ri.conicet.gov.ar:11336/159144instacron: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 09:35:59.209CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv A deficitary model of CDK5 does not impairs neuronal differentiation of human pluripotent stem cells
title A deficitary model of CDK5 does not impairs neuronal differentiation of human pluripotent stem cells
spellingShingle A deficitary model of CDK5 does not impairs neuronal differentiation of human pluripotent stem cells
Mucci, Sofia
CDK5
NEURONAL DIFFERENTIATION
HUMAN PLURIPOTENT STEM CELLS
title_short A deficitary model of CDK5 does not impairs neuronal differentiation of human pluripotent stem cells
title_full A deficitary model of CDK5 does not impairs neuronal differentiation of human pluripotent stem cells
title_fullStr A deficitary model of CDK5 does not impairs neuronal differentiation of human pluripotent stem cells
title_full_unstemmed A deficitary model of CDK5 does not impairs neuronal differentiation of human pluripotent stem cells
title_sort A deficitary model of CDK5 does not impairs neuronal differentiation of human pluripotent stem cells
dc.creator.none.fl_str_mv Mucci, Sofia
Isaja, Luciana
Rodríguez Varela, Maria Soledad
Ferriol Laffouillere, Sofia Lujan
Sevlever, Gustavo
Scassa, Maria Elida
Romorini, Leonardo
author Mucci, Sofia
author_facet Mucci, Sofia
Isaja, Luciana
Rodríguez Varela, Maria Soledad
Ferriol Laffouillere, Sofia Lujan
Sevlever, Gustavo
Scassa, Maria Elida
Romorini, Leonardo
author_role author
author2 Isaja, Luciana
Rodríguez Varela, Maria Soledad
Ferriol Laffouillere, Sofia Lujan
Sevlever, Gustavo
Scassa, Maria Elida
Romorini, Leonardo
author2_role author
author
author
author
author
author
dc.subject.none.fl_str_mv CDK5
NEURONAL DIFFERENTIATION
HUMAN PLURIPOTENT STEM CELLS
topic CDK5
NEURONAL DIFFERENTIATION
HUMAN PLURIPOTENT STEM CELLS
purl_subject.fl_str_mv https://purl.org/becyt/ford/3.1
https://purl.org/becyt/ford/3
dc.description.none.fl_txt_mv CDK5/P35 is a complex involved in neuronal homeostasis and de- velopment that was described as a critical player for neuronal surviv- al. Besides, its deregulation is linked with neurodegenerative pathol- ogies such as Alzheimer Disease and Parkinson Disease. For that reason, we generated a deficitary CDK5 genetic model in neurons derived from human pluripotent stem cells. For this purpose, we used CRISPR/Cas9 technology to generate human embryonic and induced pluripotent stem cells (hESCs and hiPSCs, respectively) KO-CDK5 lines. CDK5 protein expression levels were analyzed by western blot in samples obtained from clones where indels caused by CRISPR/Cas9 editing were detected by DNA sequencing. We obtained CDK5-/- clones for H9 hESCs and FN2.1 hiPSCs lines and a CDK5+/- clone for H9 hESCs line. Then, neural stem cells (NSC) were derived from the CDK5 KO clones using a commercial neural induction medium and their phenotype was validated by im- munofluorescence staining using antibodies that recognize specific lineage markers (SOX-1, SOX-2, NESTIN and PAX-6). Finally, NSC obtained from the heterozygous CDK5+/- KO H9 hESCs clone were differentiated into neurons using a 2D-based protocol and their phe- notype was validated by immunofluorescence staining of neuronal specific markers (TUJ-1 and MAP2). In conclusion, we managed to obtain NSC-neurons from CDK5-/- and CDK5+/- clones, determin- ing that CDK5 is not essential for NSC generation. Besides, neuro- nal differentiation was achieved for H9 CDK5+/- clone, indicating that the CDK5 deficiency does not impair the generation of NSC-derived neurons. This result allows us to account with a CDK5-defi- cient model to further study its participation in neuronal homeostasis dysfunctions.
Fil: Mucci, Sofia. Fundación para la Lucha contra las Enfermedades Neurológicas de la Infancia; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Isaja, Luciana. Fundación para la Lucha contra las Enfermedades Neurológicas de la Infancia; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Rodríguez Varela, Maria Soledad. Fundación para la Lucha contra las Enfermedades Neurológicas de la Infancia; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Ferriol Laffouillere, Sofia Lujan. Fundación para la Lucha contra las Enfermedades Neurológicas de la Infancia; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Sevlever, Gustavo. Fundación para la Lucha contra las Enfermedades Neurológicas de la Infancia; Argentina
Fil: Scassa, Maria Elida. Fundación para la Lucha contra las Enfermedades Neurológicas de la Infancia; Argentina
Fil: Romorini, Leonardo. Fundación para la Lucha contra las Enfermedades Neurológicas de la Infancia; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
LXVI Reunión Anual de la Sociedad Argentina de Investigación Clínica; LXIX Reunión Anual de la Sociedad Argentina de Inmunología; LIII Reunión Anual de la Asociación Argentina de Farmacología Experimental y XI Reunión Anual de la Asociación Argentina de Nanomedicinas
Argentina
Sociedad Argentina de Investigación Clínica
Sociedad Argentina de Inmunología
Asociación Argentina de Farmacología Experimental
Asociación Argentina de Nanomedicinas
description CDK5/P35 is a complex involved in neuronal homeostasis and de- velopment that was described as a critical player for neuronal surviv- al. Besides, its deregulation is linked with neurodegenerative pathol- ogies such as Alzheimer Disease and Parkinson Disease. For that reason, we generated a deficitary CDK5 genetic model in neurons derived from human pluripotent stem cells. For this purpose, we used CRISPR/Cas9 technology to generate human embryonic and induced pluripotent stem cells (hESCs and hiPSCs, respectively) KO-CDK5 lines. CDK5 protein expression levels were analyzed by western blot in samples obtained from clones where indels caused by CRISPR/Cas9 editing were detected by DNA sequencing. We obtained CDK5-/- clones for H9 hESCs and FN2.1 hiPSCs lines and a CDK5+/- clone for H9 hESCs line. Then, neural stem cells (NSC) were derived from the CDK5 KO clones using a commercial neural induction medium and their phenotype was validated by im- munofluorescence staining using antibodies that recognize specific lineage markers (SOX-1, SOX-2, NESTIN and PAX-6). Finally, NSC obtained from the heterozygous CDK5+/- KO H9 hESCs clone were differentiated into neurons using a 2D-based protocol and their phe- notype was validated by immunofluorescence staining of neuronal specific markers (TUJ-1 and MAP2). In conclusion, we managed to obtain NSC-neurons from CDK5-/- and CDK5+/- clones, determin- ing that CDK5 is not essential for NSC generation. Besides, neuro- nal differentiation was achieved for H9 CDK5+/- clone, indicating that the CDK5 deficiency does not impair the generation of NSC-derived neurons. This result allows us to account with a CDK5-defi- cient model to further study its participation in neuronal homeostasis dysfunctions.
publishDate 2021
dc.date.none.fl_str_mv 2021
dc.type.none.fl_str_mv info:eu-repo/semantics/publishedVersion
info:eu-repo/semantics/conferenceObject
Reunión
Journal
http://purl.org/coar/resource_type/c_5794
info:ar-repo/semantics/documentoDeConferencia
status_str publishedVersion
format conferenceObject
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/159144
A deficitary model of CDK5 does not impairs neuronal differentiation of human pluripotent stem cells; LXVI Reunión Anual de la Sociedad Argentina de Investigación Clínica; LXIX Reunión Anual de la Sociedad Argentina de Inmunología; LIII Reunión Anual de la Asociación Argentina de Farmacología Experimental y XI Reunión Anual de la Asociación Argentina de Nanomedicinas; Argentina; 2021; 1-5
CONICET Digital
CONICET
url http://hdl.handle.net/11336/159144
identifier_str_mv A deficitary model of CDK5 does not impairs neuronal differentiation of human pluripotent stem cells; LXVI Reunión Anual de la Sociedad Argentina de Investigación Clínica; LXIX Reunión Anual de la Sociedad Argentina de Inmunología; LIII Reunión Anual de la Asociación Argentina de Farmacología Experimental y XI Reunión Anual de la Asociación Argentina de Nanomedicinas; Argentina; 2021; 1-5
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://medicinabuenosaires.com/revistas/vol81-21/s3/Mv81s3.pdf
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.coverage.none.fl_str_mv Nacional
dc.publisher.none.fl_str_mv Fundación Revista Medicina
publisher.none.fl_str_mv Fundación Revista Medicina
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_ 1844613125438963712
score 13.070432

Publicaciones similares