Publicação
Identification of the proteomes of C-Kit and SCA-1 expressing populations of mice cardiac stem cells
| Resumo: | Heart failure and other heart diseases are the most important death causes in developed countries. When the heart fails, it heals by scar formation, which compromises its normal ability to pump sufficient blood to meet the metabolic demands of the body. As the heart was usually viewed as a post mitotic organ, the renewal of its tissue by endogenous cells was considered non-existent. However, recent studies in which small dividing cells with stem cell surface marker were found in the heart and identified as Cardiac Stem Cells (CSCs), changed this view and gave a new perspective in therapeutic approaches to heart related diseases. As CSCs are a recent discovery, it is essential to study and characterize these cells to better understand the way in which they regenerate the heart damaged tissue, the molecular factors involved in their mobilization and differentiation and the molecular factors produced by these cells. In the present work, the membrane and nucleus of CSCs expressing the stem cell markers Sca-1 and c-Kit were isolated using differential centrifugation, and the proteome of this nuclear and membrane fractions, along with a whole cell sample for the c-kit+ CSCs was analyzed by Peptide Mass Fingerprint, using MALDI ionization coupled with FT-ICR detection, which allowed an increase in sensitivity and resolution. 122 proteins were identified, 48 for the c-kit+ CSCs and 74 for the Sca-1+ population. Uncharacterized proteins were found in both kinds of cells, along with proteins involved in the proliferation pathways common to both populations, as is the case of the Protein Chibby Homolog 1, or specific of c-kit+ population (Alpha Enolase) or to the Sca-1+ population (47 kDa heat shock protein). Other differences were found and discussed. This work is the first attempt at the unraveling of the proteome of Cardiac Stem Cells. |
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| Autores principais: | Gomes, Adriana Andreia Silva Pires |
| Assunto: | Proteomics FACS isolation of CSCS Cardiac stem cells Membrane proteins Nuclei proteins Teses de mestrado - 2010 |
| Ano: | 2010 |
| País: | Portugal |
| Tipo de documento: | dissertação de mestrado |
| Tipo de acesso: | acesso aberto |
| Instituição associada: | Universidade de Lisboa |
| Idioma: | inglês |
| Origem: | Repositório da Universidade de Lisboa |
| Resumo: | Heart failure and other heart diseases are the most important death causes in developed countries. When the heart fails, it heals by scar formation, which compromises its normal ability to pump sufficient blood to meet the metabolic demands of the body. As the heart was usually viewed as a post mitotic organ, the renewal of its tissue by endogenous cells was considered non-existent. However, recent studies in which small dividing cells with stem cell surface marker were found in the heart and identified as Cardiac Stem Cells (CSCs), changed this view and gave a new perspective in therapeutic approaches to heart related diseases. As CSCs are a recent discovery, it is essential to study and characterize these cells to better understand the way in which they regenerate the heart damaged tissue, the molecular factors involved in their mobilization and differentiation and the molecular factors produced by these cells. In the present work, the membrane and nucleus of CSCs expressing the stem cell markers Sca-1 and c-Kit were isolated using differential centrifugation, and the proteome of this nuclear and membrane fractions, along with a whole cell sample for the c-kit+ CSCs was analyzed by Peptide Mass Fingerprint, using MALDI ionization coupled with FT-ICR detection, which allowed an increase in sensitivity and resolution. 122 proteins were identified, 48 for the c-kit+ CSCs and 74 for the Sca-1+ population. Uncharacterized proteins were found in both kinds of cells, along with proteins involved in the proliferation pathways common to both populations, as is the case of the Protein Chibby Homolog 1, or specific of c-kit+ population (Alpha Enolase) or to the Sca-1+ population (47 kDa heat shock protein). Other differences were found and discussed. This work is the first attempt at the unraveling of the proteome of Cardiac Stem Cells. |
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