Guided neural growth and axon pathfinding

2PP-DLW was introduced as an alternative approach to control various aspects in-vitro neuronal growth using mouse hippocampal neurons. Control over neuronal adhesion sites and neurite growth are particularly important in controlled in vitro neuronal networks that aim to understand the complex procedure of memory and the learning process. Prestructured cell environments featuring three-dimensional physical constraints proofed to be functional in neuron adhesion control and guided neurite growth within narrow channels interconnecting cell adhesion sites. Neurons readily aligned their processes along the desired paths and neurite extensions covered distances of more than 100 μm. Ormocomp biocompatibility, as well as the 2PP-DLW process and the chemical processing that comes along with it were found to satisfy neuronal demands regarding cleanness and cytotoxicity. The prestructured environment shows promising cell growth and guiding behavior with neurons readily aligning their growth along the channels.

Figure 1: Left: Fluorescence microscope image of mouse hippocampal neurons cultured on a sample featuring an array of cubes, fabricated by 2PP-DLW from Ormocomp. Glia cells (green, Phalloidin-Alexa 488 (actin marker)) form a carpet-like structure and grow without restrictions over the whole area, including the Ormocomp pattern (blue, autofluorescence at 561 nm excitation wavelength). Middle, right: Fluorescence microscope image of mouse hippocampal neurons cultured on a sample featuring walls defining channels and squares, fabricated by 2PP-DLW from Ormocomp. Neurites (red, MAP 2) are clearly guided over distances greater than 100 μm.