Ex vivo Culture of Mouse Embryonic Skin and Live-imaging of Melanoblast Migration

The overall goal of the following experiment is to understand the behavior of the Melano blasts population as it migrates s doso laterally through the developing embryonic epidermis. This is achieved by dissecting skin from E 14.5 mouse embryos to allow ex vivo organotypic culture. As a second step, the dissected skin is mounted in a custom built culture apparatus, which allows culture at an eliquid interface.

Next, the chamber is mounted onto the stage of a confocal microscope to image the migrating melano blasts population. Imaging the six embryonic skin cultures in parallel over an 18 hour period demonstrates the migration of Mel Oblasts as well as the development of primary hair follicles. The main advantage of this technique over older methods to visualize Mel Oblasts like immunohistochemistry in seizure hybridization, or staining lactase, is that we can do live imaging and so view individual cells to assay migration and proliferation over time.