| AVS 55th International Symposium & Exhibition | |
| Biological, Organic, and Soft Materials Focus Topic | Thursday Sessions |
| Session BO+NS+BI+NC-ThA |
| Session: | Biological and Molecular Applications of Nanostructures |
| Presenter: | L. Sirghi, JRC, Ispra, Italy |
| Authors: | L. Ceriotti, JRC, Ispra, Italy L. Buzanska, JRC, Ispra, Italy and Polish Academy of Science H. Rauscher, JRC, Ispra, Italy I. Mannelli, JRC, Ispra, Italy L. Sirghi, JRC, Ispra, Italy D. Gilliland, JRC, Ispra, Italy M. Hasiwa, JRC, Ispra, Italy F. Bretagnol, JRC, Ispra, Italy A. Ruiz, JRC, Ispra, Italy S. Bremer, JRC, Ispra, Italy S. Coecke, JRC, Ispra, Italy P. Colpo, Joint Research Center, IHCP, Italy F. Rossi, JRC, Ispra, Italy |
| Correspondent: | Click to Email |
In this work we fabricated and characterized microarrays of proteins of the extra cellular matrix (ECM) for stem cells adhesion studies. Plasma deposited poly(ethylene) oxide (PEO-like) film coated glass slides has been chosen for its dual properties, being protein and cell repellent in wet conditions and protein adhesive in dried conditions. The microarrays were created by direct microspotting of the proteins on the PEO films with optimized printing buffer by using a non-contact printing technology. The stability and the quality of the spots of fibronectin used as model protein were assessed by Time of Flight- Secondary Ion Mass Spectrometry (ToF-SIMS) and ellipsometry was used to determine the amount of protein immobilized on each spot after rinsing of the substrate with water. It was found that when fibronectin is spotted at a concentration higher than 84 µmg/ml, the protein forms a monolayer with a density of 112 ± 4 ng/cm2 with a low surface coverage but quite regular spatial distribution as confirmed by Atomic Force Microscopy (AFM) measurements. The active conformation of the spotted fibronectin as a function of the spotted concentration was verified by performing an immunoassay with antibodies specific for the fibronectin RGD sequence by Surface Plasmon Resonance (SPR) imaging. Human Umbilical Cord Blood Neural Stem Cells (HUCB-NSCs) were cultured on different ECM protein arrays (fibronectin, laminin, collagen I, collagen III and collagen V) showing a protein type and concentration dependent adhesion and growth on the micro-spots. No cells were found in-between the spots thanks to the anti adhesive properties of the PEO-like film. The cell nuclei were stained for cell counting and preliminary specific cell staining was performed to evaluate the differentiation stage of HUCB-NSCs on fibronectin spots. The array platform developed in this study provides a promising approach to investigate in a high throughput manner how insoluble factors patterned on the surface influence stem cell adhesion and development.