Aislamiento y caracterización de células madre / progenitoras y tumorales humanas: potencial terapéutico e implicaciones en carcinogénesis
- Autores: Manuel Picón Ruiz
- Directores de la Tesis: Macarena Perán (codir. tes.), Juan Antonio Marchal Corrales (codir. tes.)
- Lectura: En la Universidad de Granada ( España ) en 2012
- Idioma: español
- Tribunal Calificador de la Tesis: David Toth (presid.), Mariana Fernández Cabrera (secret.), Maria Isabel Nuñez Torres (voc.), Jose Luis Garcia Perez (voc.), Sergio Ruíz Macías (voc.)
- Materias:
- Enlaces
- Tesis en acceso abierto en: DIGIBUG
- Resumen
Stem cells (SC) and cancer stem cells (CSC) represent two cellular populations that share many similarities but, at the same time, are very different. Both are capable to long-term self-renewal and differentiation; however, their biological functions are totally opposite. SC are involved in tissue homeostasis and regeneration, representing a potent therapeutic tool for regenerative medicine. On the other hand, CSC are related with cancer progression and metastasis, being responsible of resistance to anti-cancer treatment and cancer recurrence. Hence, the correct isolation and characterization of these cell populations is essential and may have important clinical implications.
Endothelial progenitor cells (EPC) represent a relatively rare cell population, and expansion of sufficient cell numbers remains a challenge. Here we proposed a simplified and efficient method to obtain multipotent endothelial-like cells (ME-LC) from human adipose-derived stem cells (hASC) by a serial of culture stages. ME-LC population displayed increased expression levels of endothelial and hematopoietic lineage markers but no signs of mature endothelial cell differentiation. ME-LC formed capillary-like structures in vitro, secreted increased levels of stromal derived factor-1 (SDF-1), and showed ability to migrate attracted by pro-angiogenic cytokines. Importantly, ME-LC retain the capacity to proliferate extensively and differentiate into cardiomyocyte-like cells. This methodology could represent a new useful approach for cell-based therapies in cardiovascular regenerative medicine.
In this study we present an extensive evaluation of different methodologies developed to isolate and characterize CSC in two cancer cell models. Cell isolation based on specific cell surface markers expression, high aldehyde dehydrogenase (ALDH) activity and/or low adherence to plastic surface were used to isolate CSC from breast (BCSC) and colon cancer (CCSC) cell lines. We selected CSC subpopulations with high tumour markers expression, increased self-renewal, clonogenicity ability, and/or enhanced tumorigenic potential in vivo. Isolated breast CD44+/CD24low+ cells showed a phenotype more characteristic of stem cell-like than CD44+/CD24neg. In addition, we demonstrated the role of different microenviromental factors such as pro-inflammatory and pro-angiogenic cytokines in the adquisition of stem cell-like properties. Finally, we evaluated the effect of conventional and CSC-target therapies showing that strategies targeting Notch pathway by inhibition gamma-secretase activity have been shown more effective for BCSC treatment than conventional therapies based in the use of Paclitaxel.
