Method: 3-D collagen constructs (75x25x15 mm) were seeded with primary muscle precursor cells (MPC) and dorsal root ganglion cells-E7-10 (DRGs) or motor neurons, or a combination of both. Constructs were tethered at either end to develop an endogenous static load and allowing the re-alignment of cells parallel to lines of stress. This promoted formation of syncitial myotubes. Gene expression of myogenin and nAChR-epsilon were quantitated. Sucessful motorneurone isolation allowed us to seed these with MPCs within the the same type of 3-D construct. Immunostaining was used to image samples of the second model.
Results: MPC seeded constructs developed syncitial myotubes. DRG co-cultures demonstrated cell viability and alignment over the culture periods. Gene expression showed overall increases in both myogenin (0.26±0.03 for co-cultures vs. 0.08±0.01 for MPC alone) and nAChR-epsilon (0.85±0.26 for the co-cultures vs. 0.41±0.11 for MPC alone) mRNA expression when compared to single culture only.
Conclusion: Two chimaeric 3-D co-culture systems have been created with a cellular nerve-component.