Use of weak DC electric fields to rapidly align mammalian cells

Objective.The ability to modulate cell morphology has clinical relevance in regenerative biology. For example, cells of the skeletal muscle, peripheral nerve and vasculature have specific oriented architectures that emerge from unique structure-function relationships. Methods that can induce similar cell morphologiesin vitrocan be of use in the development of biomimetic constructs for the repair or replacement of damaged tissues. In this work, we demonstrate that direct current (DC) electric fields (EFs) can be used as a tool to globally align cell populationsin vitro. Approach.Using a 2D culture chamber system, we were able to quickly (within hours) align Schwann cells at different culture densities with an application of steady EFs at 200-500 mV mm^-1.Main results.Cellular alignment was perpendicular to the field vector and varied proportionately as a function of field magnitude. In addition, the degree of cellular alignment was also dependent on cellular density. Even well-established Schwann cell monolayers were responsive to the applied DC fields with cells retracting parallel oriented processes (with respect to the imposed field) and re-extending them along the perpendicular axis. When the DC field was removed, monolayers retained the aligned morphology for many days afterwards, likely due to contact inhibition. We further show the method is applicable to other field-responsive cells, such as 3T3 fibroblasts.Significance.The patterned cells provided nanoscale haptotactic cues and can be subsequently used as a basal layer for co-culturing or manipulated for other applications. DC fields represent a rapid, simple, and efficient technique compared to other cell patterning methods such as substrate manipulation.