title: Histological characterization and quantification 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 beneficial effects of cell grafting on various diseases or traumata of the central nervous system (CNS). In this chapter, we describe a histological workflow to characterize and quantify cellular events following neural and fibroblast(-like) stem cell grafting in healthy and demyelinated CNS tissue. First, we provide standardized protocols to isolate and culture eGFP(+) neural and fibroblast(-like) stem cells from embryonic mouse tissue. Second, we describe flow 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 workflow for the quantitative histological analysis of cell graft survival and endogenous astroglial and microglial responses is provided.
authors.affiliation: Laboratory of Experimental Hematology, Vaccine and Infectious Disease Institute (Vaxinfectio), University of Antwerp, Campus Drie Eiken (CDE-S6.51), Universiteitsplein 1, 2610, Antwerp (Wilrijk), Belgium.
authors.forename: Jelle
authors.initials: J
authors.lastname: Praet
authors.forename: Eva
authors.initials: E
authors.lastname: Santermans
authors.forename: Kristien
authors.initials: K
authors.lastname: Reekmans
authors.forename: Nathalie
authors.initials: N
authors.lastname: de Vocht
authors.forename: Debbie
authors.initials: D
authors.lastname: Le Blon
authors.forename: Chloé
authors.initials: C
authors.lastname: Hoornaert
authors.forename: Jasmijn
authors.initials: J
authors.lastname: Daans
authors.forename: Herman
authors.initials: H
authors.lastname: Goossens
authors.forename: Zwi
authors.initials: Z
authors.lastname: Berneman
authors.forename: Niel
authors.initials: N
authors.lastname: Hens
authors.forename: Annemie
authors.initials: A
authors.lastname: Van der Linden
authors.forename: Peter
authors.initials: P
authors.lastname: Ponsaerts
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D064987:Cell- and Tissue-Based Therapy; D003711:Demyelinating Diseases / Q000628:therapy*; D004195:Disease Models, Animal; D005260:Female; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism; D005434:Flow Cytometry; D015870:Gene Expression; D017930:Genes, Reporter; D006085:Graft Survival; D007150:Immunohistochemistry; D051379:Mice; D058953:Neural Stem Cells / Q000166:cytology* / Q000378:metabolism; D044968:Regenerative Medicine; D033581:Stem Cell Transplantation; D019076:Transgenes