Functional Stem Cell Integration Assessed by Organotypic Slice Cultures

Re-formation or preservation of functional, electrically active neural networks has been proffered as one of the goals of stem cell-mediated neural therapeutics. A primary issue for a cell therapy approach is the formation of functional contacts between the implanted cells and the host tissue. Therefore, it is of fundamental interest to establish protocols that allow us to delineate a detailed time course of grafted stem cell survival, migration, differentiation, integration, and functional interaction with the host. One option for in vitro studies is to examine the integration of exogenous stem cells into an existing active neuronal network in ex vivo organotypic cultures. Organotypic cultures leave the structural integrity essentially intact while still allowing the microenvironment to be carefully controlled. This allows detailed studies over time of cellular responses and cell-cell interactions, which are not readily performed in vivo. This unit describes procedures for using organotypic slice cultures as ex vivo model systems for studying neural stem cell and embryonic stem cell engraftment and communication with CNS host tissue. Curr. Protoc. Stem Cell Biol. 23:2D.13.1-2D.13.26. © 2012 by John Wiley & Sons, Inc. Keywords: organotypic culture; striatum; brainstem; roller drum; Stoppini; neural stem cells; engraftment; transplantation; integration; interaction; neuroprotection