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Stem cell technology provides a new tool for better understanding the mechanisms involved in compound-induced adverse reactions of the organism, which particularly applies to the fi eld of developmental neurotoxicity. Human Umbilical Cord Blood Neural Stem Cell (HUCB-NSC) line is a model system where key neurodevelopmental processes were investigated by conventional and emerging techniques. The advantage of the HUCB-NSC line is that cells are of human origin, non-transformed and can be cultured/harvested at different developmental stages. In this report emerging nano/micro-technologies were used to create biofunctional micropatterns and multi electrode array chips for the detection of cell behaviour and vulnerability to toxic compounds. Micropatterned surfaces were produced by a spatial arrangement of different functional domains. This included a nano/micro-fabrication technique like contact printing in order to create a pattern of separated or interconnected polypeptide spots directing cell growth and differentiation. Another approach was to create protein microarrays by piezoelectric (non contact) deposition of extracellular matrix proteins. Such a method allows defi ned active areas to be produced on the same platform and enables unambiguous access to cell behavior on different protein types and concentrations. HUCB-NSC were shown to adhere and differentiate on microarray platforms in a protein type, concentration and cell density dependent manner. Sensor and omics techniques applied to HUCB-NSC included measurements of electrical activity using multielectrode array chips and metabolite profi ling by mass spectrometry. Spontaneous electrical fi eld potentials and the protein composition of tested cells were sensitive to neurotoxic treatments in a developmental stage specifi c manner.
Bio-functionalized surfaces were prepared to study the adherence and differentiation capacity of neural stem cells derived from human umbilical cord blood (HUCB-NSC). Cell growth platforms containing arranged arrays of adhesive molecules were created by microcontact printing on a biologically inert surface. Biomolecules used to prepare microarray platforms included the extracellular matrix protein fibronectin and the polyaminoacid poly-L-lysine. HUCB-NSC plated on microplatforms at various serum conditions showed serum and molecule type dependent capacity for adhesion and differentiation. Poly-L-lysine allowed the maintenance of stem-like non differentiated cells attached to the surface, whereas fibronectin promoted spreading and neural commitment. Serum deprivation did not influence the attachment of HUCB-NSC to fibronectin, but significantly enhanced the attachment to poly-L-lysine and promoted dBcAMP induced neuronal differentiation. A bio-pattern of squares with interconnecting lines was used to guide neuronal differentiation by directing cell protrusion outgrowth. Tailoring the geometry of the bio-pattern enabled directing and monitoring of the neural stem cells development in the large scale multiparameter biotests.
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