Organotypic Slice Culture of Salamander Limbs: Identification and Investigation of Nerve Derived BMP2 in an Organotypic In Vitro Assay

Author(s): Lehrberg, Jeffrey | Advisor(s): Gardiner, David M | Abstract: Salamander limb regeneration is a nerve-dependent process. In the absence of nerves, blastema cells (i.e. cells that give rise to the new structure) will not proliferate. A major question in regeneration biology is how the nerve exerts its growth promoting effect on blastema cells. To better understand the cellular and molecular signals responsible for growth and pattern formation during regeneration, I have adapted the use organotypic slice culture for axolotl blastemas. Using microarray, I have identified BMP2 as a possible nerve derived factor responsible for growth and proliferation during regeneration. Here I will demonstrate how organotypic slice cultures are responsive to culture conditions and the candidate neurotrophic factor BMP2 in both terms of levels of proliferation and gene expression.