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Doctoral Thesis
DOI
10.11606/T.87.2008.tde-05012009-115547
Document
Author
Full name
Ana Carolina Souza Ramos de Carvalho
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2008
Supervisor
Committee
Moreira Filho, Carlos Alberto (President)
Bendit, Israel
Dualibi, Monica Talarico
Menck, Carlos Frederico Martins
Ribeiro, Andreza Alice Feitosa
Title in Portuguese
Estudo da proliferação e diferenciação de células-tronco hematopoéticas provenientes de sangue de cordão umbilical na presença e ausência de mitógenos.
Keywords in Portuguese
Células-tronco
Cloreto de lítio
Cordão umbilical
Estradiol
Expressão gênica
Proliferação celular
Abstract in Portuguese
Células tronco hematopoéticas (CTH) de sangue de cordão umbilical (SCU) possuem grande potencial em terapia celular. Mesmo sendo bem caracterizadas quanto às suas propriedades funcionais e fenotípicas, a regulação da auto-renovação de CTH e os genes envolvidos são pouco conhecidos. Investigou-se através da curva de crescimento, ensaio clonogênico e citometria de fluxo, a expansão e diferenciação de CTH cultivadas sem e com suplementação dos mitógenos estradiol e LiCl. A expressão da subunidade da telomerase teve aumento significativo em todas as condições, bem como a expressão de Nanog e Oct4 relacionados a pluripotência e auto-renovação. Observou-se também a expressão de Nanog, Oct4, Sox2 e FoxD3 em células CD133, células CD3 de sangue periférico e células de colônias hematopoéticas. Concluiu-se que o meio sem suplementação já é suficiente na expansão de CTH, mantendo suas características, relacionadas à proliferação, auto-renovação e pluripotência celular.
Title in English
Proliferation and differentiation study of hematopoetic stem cells from umbilical cord blood in the presence and absence of mitogens.
Keywords in English
Cell proliferation
Cord blood
Estradiol
Gene expression
Lithium chlorite
Stem cells
Abstract in English
Hematopoietic stem cells (HSC) from umbilical cord blood (UCB) have a great potencial for hematopoietic reconstitution. Although these stem cells have been well characterized by their functional and fenotipics properties, self-renewal regulation and genes involved are still unknown. Analyses of cell growth, clonogenic assay and flow cytometry revealed the expansion and differentiation of HSC grown in medium with or without suplementation of the mitogens estradiol and LiCl. Expression of the subunit of telomerase increased in all treatments. As well as the expression of Nanog and Oct4, related to plutipotency and self-renewal. Nanog, Oct4, Sox2 and FoxD3 expression was also high in CD133 cells, in CD3 cells from peripherical blood and in clonogenic assay derived cells. Conclusion: medium without the suplementation is sufficient for expansion of HSC, keeping their characteristcs, realted to proliferation, self-renewal and pluripotency.
 
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Publishing Date
2009-02-16
 
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