Microcontact printing and microspotting as methods for direct protein patterning on plasma deposited polyethylene oxide: application to stem cell patterning
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Two methods for protein patterning on antifouling surfaces have been applied to analyze the density and bioactivity of the proteins after deposition. Microcontact printing has been used as a technique to transfer fibronectin through conformal contact, while piezoelectric deposition has been employed as a non-contact technique for producing arrays of fibronectin (FN). Plasma deposited polyethylene oxide-like (PEO-like) films have been used as non-fouling background to achieve the bioadhesive/biorepellent surface contrast. Both patterning methods allow the direct fabrication of protein arrays on a non-fouling substrate, and the subsequent formation of a pattern of stem cells by cell attachment on the arrayed substrates. Microcontact printing produced fully packed homogeneous fibronectin patterns, much denser than microspotted patterns. Both printing and spotting technologies generated functional protein arrays, their bioactivity being primarily modulated by the density of the deposited protein layer. Optimization of the FN parameters used for deposition has lead to the achievement of high-quality microarrays with large population of neural stem cells immobilized in the patterns in serum-free conditions, where cells exhibit a more homogeneous starting population and factors influencing fate decisions can be more easily tracked. The immunorecognition of fibronectin targeted antibodies, as well as the cell density, increase with the protein density up to a saturation point. Over 100 ng/cm2 of fibronectin on the surface leads to a decrease in the number of attached cells and a raise of cell spreading.
KeywordsMicropatterning Surface modification Protein Cell adhesion Stem cells
This project has been financed by the European Commission Joint Research Centre Actions “NanoBiotechnology for Health” and “Validation for Consumer Products”, as well as by the Polish Ministry of Scientific Research and Higher Education grants No: 2211/B/P01/2010/38 and 5978/B/PO1/2010/38. The authors thank Ezio Parnisari for the PEO depositions and for the technical support received from Thierry Martin, Giovanni Maselli and Simone Malfara.
Time-lapse recording of HUCB-Neural Stem Cells culture on FN patterns of squares connected with lines. Arrow indicates a process of interkinetic nuclear migration (MPEG 6567 kb)
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