Authors: M. Fischer, M. Klett, U. Fernekorn, C. Augspurger, A. Schober
Affilation: Technische Universität Ilmenau, Germany
Pages: 227 - 230
Keywords: co-culture system, multi-electrode-array, neuro-muscular communication
Development of a sensor-connected in vitro neuromuscular co-culture system may provide a useful tool for interpreting mutual signal transfers between the cell species and for future applications in drug screening. A glass-based, autoclavable, micro fluidic culturing system consisting of two chambers connected by 50µm micro capillaries with a diameter of 5 µm was designed. Micro capillaries were generated by two technological approaches. First reactive ion etching was used for structuring the glass capillaries as well as a subsequent bonding to a glass carrier with electrodes by using a UV-activatable adhesive. As second approach a structured SU8-layer was used simultaneously for generating the capillaries and as an intermediate layer for bonding. Chambers and fluidic luer connectors were realized by means of a cost effective micro-sandblasting process. The co-culture system was combined with a commercially available multi electrode array. This setup allows stimulation of adherent neuronal cells in one chamber and measurement of action potentials induced in myotubes derived from the myogenic C2C12 cell line in the other. Neuronal processes growing through micro capillaries were discriminated between axons and dendrites by immuno-fluorescence staining against microtubule-associated protein and growth associated protein. This hybride technology represents a new approach for electrophysiological recordings.