Single Stem Cell Positioning on Polylysine and Fibronectin Microarrays

Abstract

Arrays of human umbilical cord blood-neural stem cells have been patterned in high density at single cell reso- lution. Pre-patterns of adhesive molecules, i.e. fibronectin and poly-L-lysine, have been produced on anti-adhesive poly (ethylene) oxide films deposited by plasma-enhanced chemical vapour deposition, which prevents cell adsorption. The structures consisted of adhesive squares and lines with 10m lateral dimensions, which correspond approximately to the size of one cell nucleus, separated by 10m anti-adhesive gap. The stem cells cultured on these platforms redistribute their cytoplasm on the permitted areas. Spherical cells were deposited on the square patterns in a single cell mode, while on the lines they spread longitudinally; the extent of elongation being dependent on the specific (fibronectin) or non-specific (poly-L-lysine) attachment biomolecule. The cell patterns were retained up to 12 days, which will be useful for recording statistical data of individual chronic responses to chemical, physical or physiologically relevant stimuli.