Determinação da atuação dos RTKs na neuroindução e neurogênese de células-tronco adultas de tecido de cordão umbilical humano

Detalhes bibliográficos
Autor(a) principal: Molina, Rachel Dias
Data de Publicação: 2015
Tipo de documento: Dissertação
Idioma: por
Título da fonte: Biblioteca Digital de Teses e Dissertações da PUC_RS
Texto Completo: http://tede2.pucrs.br/tede2/handle/tede/6311
Resumo: Introduction : The cellular and physiological changes that accompany the aging process has pronounced effects in neurodegenerative diseases. Factors that produce neurodegeneration do so through different processes that culminate in a common signaling pathway cascade. Stem cells communicate via cellular signaling mechanisms, which is part of a complex system which governs and coordinates activities and cell functions. Understanding the mechanisms and forms of intercellular communication and their relationship in the activation of signaling cascades from the surface receptor to the cell nucleus during neurogenesis, will answer some pertinent questions to the mechanisms of action of stem cells in repair central nervous system. Objective : This study aimed to determine which RTKs being activated during neurogenesis of adult stem cells from human umbilical cord tissue. Methods : Adult stem cells were obtained from human umbilical cord tissue (n = 3) and subjected to specific inducers in neurodifferentiation means. The kit Human Phospho-RTK Array was used for evaluating the phosphorylation of the RTK and the expression determined by realtime PCR. Results : The mesenchymal stem cells obtained from umbilical cord tissue were confirmed by differentiation induction in adipocytes and osteocytes cells. The neurodifferentiation was confirmed by expression of genes that are expressed during neurogenesis in cells and neural cells, as well as immunocytochemistry technique markings which showed nuclear (DAPI) and cell processes (Fluoropan). During the neuroinduction processes and neurodifferentiation the phosphorylation levels of RTKs were variable, with no clear pattern. The Ryk receptor showed no significant changes in their levels of phosphorylation of the three samples studied and the different stages of differentiation, ie, mesenchymal, neuroinductions and neurodifferentiates cells. Conclusions : The tyrosine kinases are actively being phosphorylated during neuroinduction and neurogenesis, with varying standards between the different stages and between different samples, reflecting the variability of human adult stem cells.
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spelling Machado, Denise Cantarelli294.669.110-15018.504.620-70Molina, Rachel Dias2015-09-16T22:49:16Z2015-08-03http://tede2.pucrs.br/tede2/handle/tede/6311Introduction : The cellular and physiological changes that accompany the aging process has pronounced effects in neurodegenerative diseases. Factors that produce neurodegeneration do so through different processes that culminate in a common signaling pathway cascade. Stem cells communicate via cellular signaling mechanisms, which is part of a complex system which governs and coordinates activities and cell functions. Understanding the mechanisms and forms of intercellular communication and their relationship in the activation of signaling cascades from the surface receptor to the cell nucleus during neurogenesis, will answer some pertinent questions to the mechanisms of action of stem cells in repair central nervous system. Objective : This study aimed to determine which RTKs being activated during neurogenesis of adult stem cells from human umbilical cord tissue. Methods : Adult stem cells were obtained from human umbilical cord tissue (n = 3) and subjected to specific inducers in neurodifferentiation means. The kit Human Phospho-RTK Array was used for evaluating the phosphorylation of the RTK and the expression determined by realtime PCR. Results : The mesenchymal stem cells obtained from umbilical cord tissue were confirmed by differentiation induction in adipocytes and osteocytes cells. The neurodifferentiation was confirmed by expression of genes that are expressed during neurogenesis in cells and neural cells, as well as immunocytochemistry technique markings which showed nuclear (DAPI) and cell processes (Fluoropan). During the neuroinduction processes and neurodifferentiation the phosphorylation levels of RTKs were variable, with no clear pattern. The Ryk receptor showed no significant changes in their levels of phosphorylation of the three samples studied and the different stages of differentiation, ie, mesenchymal, neuroinductions and neurodifferentiates cells. Conclusions : The tyrosine kinases are actively being phosphorylated during neuroinduction and neurogenesis, with varying standards between the different stages and between different samples, reflecting the variability of human adult stem cells.Introdução : As alterações celulares e fisiológicas que acompanham o processo de envelhecimento tem efeitos pronunciados nas doenças neurodegenerativas. Os fatores que produzem neurodegeneração o fazem através de diferentes processos, que culminam em uma via comum de cascata de sinalização. As células-tronco se comunicam por mecanismos de sinalização celular, que faz parte de um complexo sistema que governa e coordena as atividades e funções celulares. Compreender os mecanismos e as formas de comunicação intercelular e a sua relação na ativação das cascatas de sinalização, desde o receptor de superfície até o núcleo celular, durante a neurogênese, permitirá responder algumas questões pertinentes aos mecanismos de ação das células-tronco no reparo do sistema nervoso central. Objetivo : Este estudo teve como objetivo determinar quais os RTKs que estão sendo ativados durante a neurogênese de células-tronco adultas de tecido de cordão umbilical humano. Metodologia : Células-tronco adultas foram obtidas de tecido de cordão umbilical humano (n=3) e submetidas a neurodiferenciação em meios indutores específicos. O kit Human Phospho-RTK Array foi utilizado para avaliação da fosforilação dos RTKs e a expressão determinada por PCR em tempo real. Resultados : As células-tronco mesenquimais obtidas do tecido do cordão umbilical foram confirmadas através da indução de diferenciação em células adipócitas e osteócitas. A neurodiferenciação foi confirmada através da expressão de genes que são expressos nas células durante a neurogênese e em células neurais, como também a técnica de imunocitoquímica que mostrou marcações nucleares (DAPI) e prolongamentos celulares (Fluoropan). Durante os processos de neuroindução e neurodiferenciação os níveis de fosforilação dos RTKs foram variáveis, sem um padrão definido. O receptor Ryk não apresentou alterações significativas nos seus níveis de fosforilação entre as três amostras estudadas e nas diferentes etapas de diferenciação, isto é, células-tronco mesenquimais, células neuroinduzidas e células neurodiferenciadas. Conclusões : Os receptores tirosina quinases estão sendo ativamente fosforilados durante a neuroindução e neurogênese, com padrões variáveis entre as diferentes etapas e entre diferentes amostras, refletindo a variabilidade das células-tronco adultas humanas.Submitted by Setor de Tratamento da Informação - BC/PUCRS (tede2@pucrs.br) on 2015-09-16T22:49:16Z No. of bitstreams: 1 474864 - Texto Completo.pdf: 4783570 bytes, checksum: 1da454a8aaea971c62e4fff0a6d841a2 (MD5)Made available in DSpace on 2015-09-16T22:49:16Z (GMT). No. of bitstreams: 1 474864 - Texto Completo.pdf: 4783570 bytes, checksum: 1da454a8aaea971c62e4fff0a6d841a2 (MD5) Previous issue date: 2015-08-03application/pdfhttp://tede2.pucrs.br:80/tede2/retrieve/163332/474864%20-%20Texto%20Completo.pdf.jpgporPontifícia Universidade Católica do Rio Grande do SulPrograma de Pós-Graduação em Gerontologia BiomédicaPUCRSBrasilInstituto de Geriatria e GerontologiaMEDICINAGERONTOLOGIAGERONTOLOGIA BIOMÉDICACÉLULAS-TRONCOCORDÃO UMBILICALDOENÇAS NEURODEGENERATIVASENVELHECIMENTOCIENCIAS DA SAUDE::MEDICINADeterminação da atuação dos RTKs na neuroindução e neurogênese de células-tronco adultas de tecido de cordão umbilical humanoinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesis44386614769531790336006006002296420844541114010-969369452308786627info:eu-repo/semantics/openAccessreponame:Biblioteca Digital de Teses e Dissertações da PUC_RSinstname:Pontifícia Universidade Católica do Rio Grande do Sul (PUCRS)instacron:PUC_RSTHUMBNAIL474864 - Texto Completo.pdf.jpg474864 - Texto Completo.pdf.jpgimage/jpeg3221http://tede2.pucrs.br/tede2/bitstream/tede/6311/4/474864+-+Texto+Completo.pdf.jpg3785bb8981cacd2b099b049533b23d13MD54TEXT474864 - Texto Completo.pdf.txt474864 - Texto Completo.pdf.txttext/plain140657http://tede2.pucrs.br/tede2/bitstream/tede/6311/3/474864+-+Texto+Completo.pdf.txt717312a49dd086b3846b6ec827ae98d9MD53ORIGINAL474864 - Texto Completo.pdf474864 - Texto Completo.pdfapplication/pdf4783570http://tede2.pucrs.br/tede2/bitstream/tede/6311/2/474864+-+Texto+Completo.pdf1da454a8aaea971c62e4fff0a6d841a2MD52LICENSElicense.txtlicense.txttext/plain; charset=utf-8610http://tede2.pucrs.br/tede2/bitstream/tede/6311/1/license.txt5a9d6006225b368ef605ba16b4f6d1beMD51tede/63112015-09-29 08:33:52.073oai:tede2.pucrs.br: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Biblioteca Digital de Teses e Dissertaçõeshttp://tede2.pucrs.br/tede2/PRIhttps://tede2.pucrs.br/oai/requestbiblioteca.central@pucrs.br||opendoar:2015-09-29T11:33:52Biblioteca Digital de Teses e Dissertações da PUC_RS - Pontifícia Universidade Católica do Rio Grande do Sul (PUCRS)false
dc.title.por.fl_str_mv Determinação da atuação dos RTKs na neuroindução e neurogênese de células-tronco adultas de tecido de cordão umbilical humano
title Determinação da atuação dos RTKs na neuroindução e neurogênese de células-tronco adultas de tecido de cordão umbilical humano
spellingShingle Determinação da atuação dos RTKs na neuroindução e neurogênese de células-tronco adultas de tecido de cordão umbilical humano
Molina, Rachel Dias
MEDICINA
GERONTOLOGIA
GERONTOLOGIA BIOMÉDICA
CÉLULAS-TRONCO
CORDÃO UMBILICAL
DOENÇAS NEURODEGENERATIVAS
ENVELHECIMENTO
CIENCIAS DA SAUDE::MEDICINA
title_short Determinação da atuação dos RTKs na neuroindução e neurogênese de células-tronco adultas de tecido de cordão umbilical humano
title_full Determinação da atuação dos RTKs na neuroindução e neurogênese de células-tronco adultas de tecido de cordão umbilical humano
title_fullStr Determinação da atuação dos RTKs na neuroindução e neurogênese de células-tronco adultas de tecido de cordão umbilical humano
title_full_unstemmed Determinação da atuação dos RTKs na neuroindução e neurogênese de células-tronco adultas de tecido de cordão umbilical humano
title_sort Determinação da atuação dos RTKs na neuroindução e neurogênese de células-tronco adultas de tecido de cordão umbilical humano
author Molina, Rachel Dias
author_facet Molina, Rachel Dias
author_role author
dc.contributor.advisor1.fl_str_mv Machado, Denise Cantarelli
dc.contributor.advisor1ID.fl_str_mv 294.669.110-15
dc.contributor.authorID.fl_str_mv 018.504.620-70
dc.contributor.author.fl_str_mv Molina, Rachel Dias
contributor_str_mv Machado, Denise Cantarelli
dc.subject.por.fl_str_mv MEDICINA
GERONTOLOGIA
GERONTOLOGIA BIOMÉDICA
CÉLULAS-TRONCO
CORDÃO UMBILICAL
DOENÇAS NEURODEGENERATIVAS
ENVELHECIMENTO
topic MEDICINA
GERONTOLOGIA
GERONTOLOGIA BIOMÉDICA
CÉLULAS-TRONCO
CORDÃO UMBILICAL
DOENÇAS NEURODEGENERATIVAS
ENVELHECIMENTO
CIENCIAS DA SAUDE::MEDICINA
dc.subject.cnpq.fl_str_mv CIENCIAS DA SAUDE::MEDICINA
description Introduction : The cellular and physiological changes that accompany the aging process has pronounced effects in neurodegenerative diseases. Factors that produce neurodegeneration do so through different processes that culminate in a common signaling pathway cascade. Stem cells communicate via cellular signaling mechanisms, which is part of a complex system which governs and coordinates activities and cell functions. Understanding the mechanisms and forms of intercellular communication and their relationship in the activation of signaling cascades from the surface receptor to the cell nucleus during neurogenesis, will answer some pertinent questions to the mechanisms of action of stem cells in repair central nervous system. Objective : This study aimed to determine which RTKs being activated during neurogenesis of adult stem cells from human umbilical cord tissue. Methods : Adult stem cells were obtained from human umbilical cord tissue (n = 3) and subjected to specific inducers in neurodifferentiation means. The kit Human Phospho-RTK Array was used for evaluating the phosphorylation of the RTK and the expression determined by realtime PCR. Results : The mesenchymal stem cells obtained from umbilical cord tissue were confirmed by differentiation induction in adipocytes and osteocytes cells. The neurodifferentiation was confirmed by expression of genes that are expressed during neurogenesis in cells and neural cells, as well as immunocytochemistry technique markings which showed nuclear (DAPI) and cell processes (Fluoropan). During the neuroinduction processes and neurodifferentiation the phosphorylation levels of RTKs were variable, with no clear pattern. The Ryk receptor showed no significant changes in their levels of phosphorylation of the three samples studied and the different stages of differentiation, ie, mesenchymal, neuroinductions and neurodifferentiates cells. Conclusions : The tyrosine kinases are actively being phosphorylated during neuroinduction and neurogenesis, with varying standards between the different stages and between different samples, reflecting the variability of human adult stem cells.
publishDate 2015
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