Protocol.pdf

Brain Research Protocols 5 (2000) 282–289 / locate/ bres Protocol Culturing neuronal cells on surfaces coated by a novel polyethyleneimine-based polymer aba, * Yaniv Bledi , Abraham J. Domb , Michal Linial Department of Biological Chemistry ,The Alexander Silberman Institute of Life Sciences ,The Hebrew University , Jerusalem 91904, Israel School of Pharmacy ,Faculty of Medicine ,The Hebrew University , Jerusalem 91120, Israel Accepted 7 April 2000 Abstract Maintaining cells in culture is essential for studying many aspects of cell biology and physiology. Cell culturing is dependent on proper anchorage of cells to the growth surfaces. For most cell lines, and especially for post-mitotic neurons, coated tissue culture plates are prerequisite for seeding. The monly used coating reagents are positively charged polymers such as poly- L-lysine or biologically puri?ed adhesive molecules such as collagen. In this report, we present a simple procedure for synthesizing and for coating cell culture surfaces. The reagent is a biologically inert hydrophobized polyethyleneimine (PEI), which provides adequate adhesive properties for cultured cell lines including those of neuronal lineage. The hydrophobized PEI is branched PEI modi?ed by octadecanyl groups bound to 2 mol% of the amino groups of the PEI. Unlike the native PEI that is water soluble, the modi?ed PEI is soluble in ethanol, and th