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Doctoral Thesis
DOI
https://doi.org/10.11606/T.46.2008.tde-29052008-144040
Document
Author
Full name
Fernando Henrique Lojudice da Silva
E-mail
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2008
Supervisor
Committee
Sogayar, Mari Cleide (President)
Carramaschi, Lygia da Veiga Pereira
Krieger, José Eduardo
Silva, Aline Maria da
Zingales, Bianca Silvana
Title in Portuguese
Identificação de genes diferencialmente expressos durante diferenciação de células-tronco e caracterização de células progenitoras mesenquimais
Keywords in Portuguese
Células-tronco
CodeLink
Cordão umbilical
Diabetes
Eucarioto
Expressão gênica
Insulina
Abstract in Portuguese
Diabetes mellitus (DM) designa um conjunto de patologias devidas à falta ou ação deficiente da insulina. O DM1 é causado por ataque auto-imune às células β-pancreáticas secretoras de insulina, enquanto DM2 é relacionado com idade e obesidade. Alotransplante de pâncreas ou de ilhotas são alternativas terapêuticas, mas escassez de órgãos e necessidade de imunossupressão são obstáculos importantes. Células-tronco, isoladas da massa interna de blastocistos e de tecidos adultos, são capazes de auto-renovação e diferenciação, podendo ser utilizadas para o tratamento de doenças. Os objetivos deste trabalho foram: a) buscar novos genes envolvidos no processo de diferenciação de células tronco em células secretoras de insulina e b) isolar e caracterizar células-tronco mesenquimais humanas. Células-tronco embrionárias murinas foram induzidas a se diferenciar em clusters similares a ilhotas. O RNA foi utilizado para sondar microarrays de DNA (CodeLink) e os genes diferencialmente expressos foram confirmados por Q-PCR, sendo possível identificar 16 novos genes associados à diferenciação. Dois tipos diferentes de células-tronco mesenquimais foram isoladas da veia (MSC) e do sangue (UCB) de cordão umbilical humano. Imunofenotipagem e caracterização molecular por Q-PCR, apontaram para a existência de dois tipos diferentes de progenitoras mesenquimais adultas no cordão umbilical humano.
Title in English
Identification of differentialy express genes during the differentiation of stem cells and characterization of mesenchymal progenitor cells
Keywords in English
CodeLink
Diabetes
Insulin
Stem cells
Umbilical cord
Abstract in English
Diabetes mellitus (DM) defines a number of pathologies caused by the lack or deficient action of the insulin hormone. DM1 is caused by an auto-immune attack to insulin secreting β-pancreatic cells, while DM2 is related to ageing and obesity. Pancreas and islet allo-transplantation constitute therapeutic alternatives, but severe organ shortage and the absolute requirement for immunossupression still constitute important obstacles. Stem cells isolated from the blastocist inner cell mass and from adult tissues are capable of self-renewal and differentiation, and may be utilized for treatment of several diseases. The objectives of this work were: a) to search for new genes involved in the process of differentiation of stem cells into insulin-secreting cells and b) to isolate and characterize mesenchymal stem cells. Murine embryonic stem cells were induced to differentiate in islet-like clusters. Total RNA was utilized to probe DNA microarrays (CodeLink) and the differentially expressed genes were confirmed by Q-PCR, with 16 new genes being identified as associated with differentiation. Two different types of mesenchymal stem cells were isolated from human umbilical cord vein (MSC) and blood (UCB). Immunophenotyping and molecular characterization by Q-PCR, pointed to the existence of two different types of progenitor mesenchymal stem cells in human umbilical cord.
 
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Tese.pdf (27.44 Mbytes)
Publishing Date
2008-06-13
 
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