Derivação de células tronco de tecido ovariano na espécie canina

• Ano de defesa: 2014
• Autor(a) principal: Trindade, Amanda Baracho [UNESP]
• Tipo de documento: Dissertação
• Tipo de acesso: Acesso aberto
• Idioma: por
• Instituição de defesa: Universidade Estadual Paulista (Unesp)
• Palavras-chave em Português: Cão; Células-tronco; Ovarios; Tecido adiposo; Marcadores biologicos; Citometria de fluxo; Stem cells
• Link de acesso: http://hdl.handle.net/11449/126360; http://www.athena.biblioteca.unesp.br/exlibris/bd/cathedra/12-08-2015/000836696.pdf

Research with canine stem cells contributes to the progress of veterinary and human cellular therapies, because the dog is often the best animal model for many hereditary and acquired human diseases. While embryonic SC have unlimited differentiation potential (i.e. pluripotent), adult-derived multipotent SC are often preferred for ease of recovery from the patient itself (i.e. autologous) and for avoiding ethical constraints of destroying embryos to obtain pluripotent SC. Recent promising studies have shown the existence SC in ovarian tissue, that have greater proliferative and differentiation potential than SC in other adult tissues. Based on this evidence, the aim of this project was to obtain stem cells isolated from ovarian tissue of adult dogs. In experiment I, we evaluated morphological and kinetic properties of collagenase-digested ovarian and adipose derived SC. Adipose derived cells yielded more cells per tissue after 72h in culture (1937 ± 592.6 vs 1771 ± 262.1), both cells had the same fibroblastic morphology. The growth curve showed that adipose derived cells started the exponential growth before the ovarian derived cells, however when the both were multiplying, the population doubling time was the same. All the derived cells types were able to form colonies, however, adipose derived cells formed more colonies than ovarian derived cells (37.08 ± 2.10 vs. 27.08 ± 1.45). Experiment II aimed at characterizing at the molecular level the SC obtained in experiment I. We evaluated the expression of mesenchymal and pluripotent markers through immunofluorescence (flow cytometry and confocal microscopy) and PCR. Both cells from expressed CD90, CD44, CD194 and CD 105 by PCR, were positive for CD44, CD90 (only the adipose tissue) and negative for CD45 and CD34 by immunocytochemistry analysis. By flow cytometry, 37.08%+-11.35% of the population of cells derived from ovarian tissue was positive for CD90, while 62.05%+-8.40% of ...