journal homepage: www.elsevier.com/locate/yexcr Available online at www.sciencedirect.com Research Article High content and high throughput screening to assess the angiogenic and neurogenic actions of mesenchymal stem cells in vitro Jennifer J. Bara a,n , Sarah Turner b , Sally Roberts b , Gareth Griffiths c , Rod Benson c , Jayesh M. Trivedi b , Karina T. Wright b a AO Research Institute Davos, Clavadelerstrasse 8, 7270 Davos Platz, Switzerland b Spinal Studies and Institute for Science and Technology in Medicine, Keele University, ARC Building, RJAH Orthopaedic Hospital, Oswestry SY10 7AG, UK c Imagen Biotech, Manchester, UK articleinformation Article Chronology: Received 26 September 2014 Received in revised form 16 December 2014 Accepted 30 December 2014 Available online 10 February 2015 Keywords: Mesenchymal stem cell High content Secretome Conditioned media Angiogenesis Neurogenesis abstract Mesenchymal stem cells are universally regarded across many fields of medicine, as one of the most promising cell types for use in cell-based therapies. Although not yet fully understood, the therapeutic effects of these cells are largely attributed to the trophic actions of growth factors and cytokines present in the cell secretome. Specifically, the angiogenic and neurogenic properties of these cells make them attractive for the repair of vascularised and innervated tissues. In this study, we investigate the effect of mesenchymal stem cell conditioned media on in vitro assays of angiogenesis and nerve growth. We describe the use of two state of the art high content and high throughput cell analysis systems and compare them against manual analysis techniques. Mesenchymal stem cell secretomes stimulated angiogenesis and nerve growth in vitro in a donor dependant manner. Levels of neuroregulin, platelet-derived growth factor-AA and glial- derived neurotrophic factor, positively correlated with the observed angiogenic effects of these cells. High content and high throughput cell analysis systems such as the ones used in this study, may provide rapid screening tools to assist not only with patient selection but the identification of predictive therapeutic markers to support clinical outcome monitoring for patients treated with stem cell therapies. & 2015 Elsevier Inc. All rights reserved. http://dx.doi.org/10.1016/j.yexcr.2014.12.019 0014-4827/& 2015 Elsevier Inc. All rights reserved. Abbreviations: BMSC, Bone marrow-derived mesenchymal stem cell; VEGF, Vascular endothelial growth factor; FGF, Fibroblast growth factor; PDGF, Platelet-derived growth factor; NRG1, Neuroregulin-1; GDNF, Glial-derived neurotrophic factor; BDNF, Brain-derived neurotrophic factor; IL-6/IL-1β, Interleukin-6, Interleukin-1β; PIGF, Placental growth factor; MMP-8/-9, Matrix metalloproteinase; MCP-1, Monocyte chemoattractant protein; DMEM F12, Dulbecco's Modified Eagle Medium: Nutrient Mixture F-12; FCS, Foetal calf serum; PBS, Phosphate buffered saline; SCF, Stem cell factor; NT4, Neurotrophin 4; DRG, Dorsal root ganglia; HUVEC, Human umbilical vein endothelial cell; SCI, Spinal cord injury n Corresponding author. Fax: þ41 81 414 22 88. E-mail address: jennifer.bara@aofoundation.org (J.J. Bara). EXPERIMENTAL CELL RESEARCH 333 (2015) 93 – 104