title: A comparison of high-content screening versus manual analysis to assay the effects of mesenchymal stem cell-conditioned medium on neurite outgrowth in vitro.
abstract: Bone marrow mesenchymal stem cells (MSCs) promote nerve growth and functional recovery in animal models of spinal cord injury (SCI) to varying levels. The authors have tested high-content screening to examine the effects of MSC-conditioned medium (MSC-CM) on neurite outgrowth from the human neuroblastoma cell line SH-SY5Y and from explants of chick dorsal root ganglia (DRG). These analyses were compared to previously published methods that involved hand-tracing individual neurites. Both methods demonstrated that MSC-CM promoted neurite outgrowth. Each showed the proportion of SH-SY5Y cells with neurites increased by ~200% in MSC-CM within 48 h, and the number of neurites/SH-SY5Y cells was significantly increased in MSC-CM compared with control medium. For high-content screening, the analysis was performed within minutes, testing multiple samples of MSC-CM and in each case measuring >15,000 SH-SY5Y cells. In contrast, the manual measurement of neurite outgrowth from >200 SH-SY5Y cells in a single sample of MSC-CM took at least 1 h. High-content analysis provided additional measures of increased neurite branching in MSC-CM compared with control medium. MSC-CM was also found to stimulate neurite outgrowth in DRG explants using either method. The application of the high-content analysis was less well optimized for measuring neurite outgrowth from DRG explants than from SH-SY5Y cells.
authors.affiliation: ISTM Keele University at the RJAH Orthopaedic Hospital, Oswestry, Shropshire, UK. Karina.Wright@rjah.nhs.uk
authors.forename: Karina T
authors.initials: KT
authors.lastname: Wright
authors.forename: Gareth J
authors.initials: GJ
authors.lastname: Griffiths
authors.forename: William E B
authors.initials: WE
authors.lastname: Johnson
mesh_terms: D000465:Algorithms; D000818:Animals; D045744:Cell Line, Tumor; D002642:Chick Embryo; D017077:Culture Media, Conditioned / Q000494:pharmacology*; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000737:chemistry*; D016501:Neurites / Q000187:drug effects*
chemical_list: D017077:Culture Media, Conditioned