| AVS 57th International Symposium & Exhibition | |
| Biomaterial Interfaces | Tuesday Sessions |
| Session BI-TuM |
| Session: | Cells on Surfaces |
| Presenter: | D. Mehn, Joint Research Centre, Italy |
| Authors: | D. Mehn, Joint Research Centre, Italy J. Nowak, Joint Research Centre, Italy L.M. Buzanska, Polish Academy of Sciences, Poland A. Ruiz, Joint Research Centre, Italy H. Rauscher, Joint Research Centre, Italy G.R. Marchesini, Joint Research Centre, Italy P. Colpo, Joint Research Centre, Italy F. Rossi, Joint Research Centre, Italy |
| Correspondent: | Click to Email |
Human umbilical cord blood originated neural stem cells (HUCB-NSC) are promising candidates for developing human cell based in vitro neurotoxicity assays. The origin of the cells makes possible to avoid species specificity extrapolation problems and avoid ethical issues compared to the embryonic stem cell research. Besides the easy cell line like culturing procedure of these cells they were shown to be able to differentiate to neuronal, asctrocytic and oligodendroglial phenotypes. This feature makes them also excellent subjects for developmental neurotoxicity studies.
The extracellular matrix compounds (ECM) used for anchoring the cells on culture dish or device surfaces may induce various ECM-dependent stem cell developmental responses mediated by integrin receptors. Investigation of stem cell - extracellular matrix interactions on biomolecule microarrays provides fundamental knowledge on neural differentiation as well as key inputs for stem cell based assay development.
The most powerful tools for microarray fabrication are microspotting and microcontact printing, offering flexibility regarding composition, concentration, spotted volumes or printed pattern geometry. Both methods are proven to be gentle enough to handle biomolecules for cell interaction studies.
In this work, the piezoelectric spotting and microcontact printing technologies were tested for rapid fabrication of biomolecule arrays on cell repellent surfaces.
Cell anchoring molecules, like extracellular matrix proteins (fibronectin and vitronectin) and poly-L-lysine (a polycationic polypeptide) were printed on anti-adhesive poly(ethylene) oxide-like films deposited by plasma-enhanced chemical vapor deposition. The microspotted and printed patterns were characterized by ellipsometry and microscopic techniques. The morphology of seeded stem cells and expression of certain differentiation marker proteins (GFAP, beta-tubulin III) were visualized by immunostaining and fluorescent microscopy, including also characterization after treatment with various concentrations of a known neurotoxic compound (MeHgCl). Toxicity results obtained on adhesive molecule spots were compared with results of a conventional neutral red uptake assay performed in coated 96 well plates.
Effect of the nature of biomolecules on cell behavior (including cell adhesion, morphology and survival) were investigated and are discussed focusing on generation long term cell-pattern stability, as well as stem cell differentiation into particular cell types.