Histological characterization and quantifi cation of cellular events following neural and fibroblast(-like) stem cell grafting in healthy and demyelinated CNS tissue

Abstract

Preclinical animal studies involving intracerebral (stem) cell grafting are gaining popularity in many laboratories due to the reported benefi cial effects of cell grafting on various diseases or traumata of the central nervous system (CNS). In this chapter, we describe a histological workfl ow to characterize and quantify cellular events following neural and fi broblast(-like) stem cell grafting in healthy and demyelinated CNS tissue. First, we provide standardized protocols to isolate and culture eGFP + neural and fi broblast(-like) stem cells from embryonic mouse tissue. Second, we describe fl ow cytometric procedures to determine cell viability, eGFP transgene expression, and the expression of different stem cell lineage markers. Third, we explain how to induce reproducible demyelination in the CNS of mice by means of cuprizone administration, a validated mouse model for human multiple sclerosis. Fourth, the technical procedures for cell grafting in the CNS are explained in detail. Finally, an optimized and validated workfl ow for the quantitative histological analysis of cell graft survival and endogenous astroglial and microglial responses is provided.