RESUMO
We report the use of self-assembled peptide (F2/S) hydrogels and cellular metabolomics to identify a number of innate molecules that are integral to the metabolic processes which drive cellular differentiation of multipotent pericyte stem cells. The culture system relies solely on substrate mechanics to induce differentiation in the absence of traditional differentiation media and therefore is a non-invasive approach to assessing cellular behavior at the molecular level and identifying key metabolites in this process. This novel approach demonstrates that simple metabolites can provide an alternative means to direct stem cell differentiation and that biomaterials can be used to identify them simply and quickly.
Assuntos
Metabolômica/métodos , Pericitos/citologia , Pericitos/transplante , Animais , Materiais Biocompatíveis/metabolismo , Capilares/citologia , Diferenciação Celular/efeitos dos fármacos , Diferenciação Celular/fisiologia , Células Cultivadas , Células Endoteliais/citologia , Humanos , Hidrogéis/química , Microvasos/citologia , Células-Tronco Multipotentes/efeitos dos fármacos , Peptídeos/química , Pericitos/metabolismo , FenótipoRESUMO
Aromatic peptide amphiphiles can form self-supporting nanostructured hydrogels with tunable mechanical properties and chemical compositions. These hydrogels are increasingly applied in two-dimensional (2D) and three-dimensional (3D) cell culture, where there is a rapidly growing need to store, grow, proliferate, and manipulate naturally derived cells within a hydrated, 3D matrix. Biogelx Limited is a biomaterials company, created to commercialize these bio-inspired hydrogels to cell biologists for a range of cell culture applications. This chapter describes methods of various characterization and cell culture techniques specifically optimized for compatibility with Biogelx products.
Assuntos
Hidrogéis/química , Peptídeos/química , Multimerização Proteica , Materiais Biocompatíveis/química , Técnicas de Cultura de Células , Hidrogéis/isolamento & purificação , Microscopia , Peptídeos/isolamento & purificação , Esferoides CelularesRESUMO
The ability to develop new and sensitive methods of biomolecule detection is crucial to the advancement of pre-clinical disease diagnosis and effective patient specific treatment. Surface enhanced Raman scattering (SERS) is an optical spectroscopy amenable to this goal, as it is capable of extremely sensitive biomolecule detection and multiplexed analysis. This perspective highlights where SERS has been successfully used to detect target biomolecules, specifically DNA and proteins, and where in vivo analysis has been successfully utilised. The future of SERS development is discussed and emphasis is placed on the steps required to transport this novel technique from the research laboratory to a clinical setting for medical diagnostics.
Assuntos
DNA/análise , Proteínas/análise , Análise Espectral Raman , DNA/química , Sondas de DNA/química , Corantes Fluorescentes/química , Imunoensaio , Nanopartículas/químicaRESUMO
We have combined the benefits of a TaqMan assay with surface enhanced Raman scattering (SERS), to generate a novel DNA detection method which provides increased sensitivity, with clear applications for disease identification through clinical testing. Target DNA detection limits by SERS were shown to be lower than conventional fluorescence detection and clinically relevant samples of methicillin-resistant Staphylococcus aureus were detected with high specificity.
Assuntos
DNA/análise , Reação em Cadeia da Polimerase , Análise Espectral Raman , Taq Polimerase/metabolismo , DNA Bacteriano/análise , Limite de Detecção , Staphylococcus aureus Resistente à Meticilina/genética , Propriedades de SuperfícieRESUMO
Developments in specific DNA detection assays have been shown to be increasingly beneficial for molecular diagnostics and biological research. Many approaches use optical spectroscopy as an assay detection method and, owing to the sensitivity and molecular specificity offered, surface enhanced Raman scattering (SERS) spectroscopy has become a competitively exploited technique. This study utilises SERS to demonstrate differences in affinity of dye labelled DNA through differences in electrostatic interactions with silver nanoparticles. Results show clear differences in the SERS intensity obtained from single stranded DNA, double stranded DNA and a free dye label and demonstrate surface attraction is driven through electrostatic charges on the nucleotides and not the SERS dye. It has been further demonstrated that, through optimisation of experimental conditions and careful consideration of sequence composition, a DNA detection method with increased sample discrimination at lower DNA concentrations can be achieved.