A comparison of high-content screening versus manual analysis to assay the effects of mesenchymal stem cell-conditioned medium on neurite outgrowth in vitro.
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.
Wright
Karina T
KT
ISTM Keele University at the RJAH Orthopaedic Hospital, Oswestry, Shropshire, UK. Karina.Wright@rjah.nhs.uk
Griffiths
Gareth J
GJ
Johnson
William E B
WE
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*
D017077:Culture Media, Conditioned