Culturing neuronal cells on surfaces coated by a novel polyethyleneimine-based polymer.
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 most commonly used coating reagents are positively charged polymers such as poly-L-lysine or biologically purified 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 modified by octadecanyl groups bound to 2 mol% of the amino groups of the PEI. Unlike the native PEI that is water soluble, the modified PEI is soluble in ethanol, and thus resistant to solubilization in biological media. The protocol of coating was optimized for tissue culture plates as well as glass surfaces and in many respects this polymer outperformed other routinely used coating reagents. Neuronal cell lines, plated on the polymer-treated surfaces are resistant to manipulations including repeated media changes and extensive washing. The advantage of coating surfaces with the developed PEI-based polymer compared to other commonly used coating reagents is discussed.
Bledi
Y
Y
Department of Biological Chemistry, The Alexander Silberman Institute of Life Sciences, The Hebrew University, 91904, Jerusalem, Israel.
Domb
A J
AJ
Linial
M
M
D000818:Animals; D002448:Cell Adhesion / Q000187:drug effects; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D005455:Fluorescent Antibody Technique; D005898:Glass; D006801:Humans; D007124:Immunoenzyme Techniques; D007668:Kidney / Q000166:cytology; D016501:Neurites / Q000187:drug effects* / Q000502:physiology; D009474:Neurons / Q000648:ultrastructure*; D016716:PC12 Cells; D011094:Polyethyleneimine / Q000737:chemistry / Q000494:pharmacology*; D011108:Polymers; D051381:Rats
D011108:Polymers; D011094:Polyethyleneimine