RESUMO
OBJECTIVE: A significant amount of recent interest has been focused on the possibility that adult human stem cells are a realistic therapeutic alternative to embryonic stem cells. Multipotent stem cells that have characteristics reminiscent of embryonic neural crest stem cells have been isolated from several postnatal tissues, including skin, gut, dental pulp and the heart, and are potentially useful for research and therapeutic purposes. However, their neurogenic potential, including their ability to produce electrophysiologically active neurons, is largely unexplored. In the present work, we investigated this issue with regard to skin-derived precursors (SKPs) and adipose-derived stem cells (ADSc) METHODS: Adult stem cells isolated from skin and from adipose tissue derived from the same adult donor were treated with epidermal growth factor (EGF) and fibroblast growth factor 2 (FGF2). Neurospheres obtained were first expanded and evaluated in term of proliferative ability, and then their neuronal differentiation potential was analysed. RESULTS: Adipose- and skin-derived neurospheres grew in suspension as spheres in the presence of the mitogens FGF2 and EGF. With this protocols, the spheres have been able to proliferate and to originate Schwann and glial-like cells. DISCUSSION: In summary, we have demonstrated in this work that multipotent adult precursor cell can be isolated and expanded from two accessible adult tissue sources: skin and adipose tissue. The work described in this paper provides the framework for our attempts to use SKPs or ADSc as autologous adult stem cell population for cell replacement and discovery research.
Assuntos
Tecido Adiposo/citologia , Células-Tronco Adultas/fisiologia , Células-Tronco Multipotentes/fisiologia , Neurônios/citologia , Neurônios/fisiologia , Pele/citologia , Tecido Adiposo/fisiologia , Adulto , Células-Tronco Adultas/citologia , Técnicas de Cultura de Células , Diferenciação Celular/fisiologia , Proliferação de Células , Separação Celular , Células Cultivadas , Fator de Crescimento Epidérmico/metabolismo , Fator 2 de Crescimento de Fibroblastos/metabolismo , Imunofluorescência , Humanos , Pessoa de Meia-Idade , Mitocôndrias/fisiologia , Células-Tronco Multipotentes/citologia , Neurogênese/fisiologia , Neuroglia/citologia , Neuroglia/fisiologia , Fenômenos Fisiológicos da Pele , Fatores de TempoRESUMO
The ideal bioartificial liver should be designed to reproduce as nearly as possible in vitro the habitat that hepatic cells find in vivo. In the present work, we investigated the in vitro perfusion condition with a view to improving the hepatic differentiation of pluripotent human liver stem cells (HLSCs) from adult liver. Tissue engineering strategies based on the cocultivation of HLSCs with hepatic stellate cells (ITO) and with several combinations of medium were applied to improve viability and differentiation. A mathematical model estimated the best flow rate for perfused cultures lasting up to 7 days. Morphological and functional assays were performed. Morphological analyses confirmed that a flow of perfusion medium (assured by the bioreactor system) enabled the in vitro organization of the cells into liver clusters even in the deeper levels of the sponge. Our results showed that, when cocultured with ITO using stem cell medium, HLSCs synthesized a large amount of albumin and the MTT test confirmed an improvement in cell proliferation. In conclusion, this study shows that our in vitro cell conditions promote the formation of clusters of HLSCs and enhance the functional differentiation into a mature hepatic population.
Assuntos
Células-Tronco Adultas/citologia , Células-Tronco Adultas/fisiologia , Diferenciação Celular/fisiologia , Hepatócitos/fisiologia , Engenharia Tecidual/métodos , Adulto , Técnicas de Cultura de Células/métodos , Proliferação de Células , Sobrevivência Celular , Células Cultivadas , Técnicas de Cocultura/métodos , Hepatócitos/citologia , Humanos , Fígado/citologia , Modelos Biológicos , Modelos TeóricosRESUMO
OBJECTIVE: In recent years, research on stem cells has been focused on the development of personalized cell-based therapies. Owing to their homing properties, adult human stem cells are a promising source of autologous cells to be used as therapeutic vehicles. Multiple potential sources for clinically useful stem and progenitor cells have been identified, including autologous and allogenic embryonic, fetal and adult somatic cells from neural, adipose and mesenchymal tissue. In the present report, we describe a simple protocol to obtain an enriched culture of adult stem cells organized in neurospheres from two post-natal tissues: skin and adipose tissue. METHODS: Adult stem cells isolated from skin and adipose tissue derived from the same adult donor were amplified under varying conditions related to the coating of the chamber slide and the presence of serum and/or growth factors, such as with EGF and FGF2. Neurospheres were then expanded and evaluated in terms of proliferation and gene expression. RESULTS: Adipose and skin derived neurospheres were comparable in size, quantity of cells and genes expressed. Cells from both types of tissue grew optimally without slide coating, in the presence of serum and with the combined addition of FGF2 and EGF. DISCUSSION: We describe a method for isolating and improving a population of multipotent adult precursor cells from the two most accessible adult tissue sources: skin and adipose tissue. This autologous adult stem cell population could be used for cell replacement or cell therapies.