title: Surpassing light-induced cell damage in vitro with novel cell culture media.
abstract: Light is extensively used to study cells in real time (live cell imaging), separate cells using fluorescence activated cell sorting (FACS) and control cellular functions with light sensitive proteins (Optogenetics). However, photo-sensitive molecules inside cells and in standard cell culture media generate toxic by-products that interfere with cellular functions and cell viability when exposed to light. Here we show that primary cells from the rat central nervous system respond differently to photo-toxicity, in that astrocytes and microglia undergo morphological changes, while in developing neurons and oligodendrocyte progenitor cells (OPCs) it induces cellular death. To prevent photo-toxicity and to allow for long-term photo-stimulation without causing cellular damage, we formulated new photo-inert media called MEMO and NEUMO, and an antioxidant rich and serum free supplement called SOS. These new media reduced the detrimental effects caused by light and allowed cells to endure up to twenty times more light exposure without adverse effects, thus bypassing the optical constraints previously limiting experiments.
authors.affiliation: Wellcome Trust-Medical Research Council Cambridge Stem Cell Institute & Department of Veterinary Medicine, University of Cambridge, Cambridge, CB2 1QR, United Kingdom. js810@medschl.cam.ac.uk.|Wellcome Trust-MRC Cambridge Stem Cell InstituteDepartment of Paediatrics, Cambridge Biomedical Campus, The Clifford Allbutt Building, Level 6, Hills Road, Cambridge, CB2 0AH, UK. js810@medschl.cam.ac.uk.
authors.forename: John H
authors.identifier: http://orcid.org/0000-0002-7385-8310
authors.initials: JH
authors.lastname: Stockley
authors.affiliation: Wellcome Trust-Medical Research Council Cambridge Stem Cell Institute & Department of Veterinary Medicine, University of Cambridge, Cambridge, CB2 1QR, United Kingdom.
authors.forename: Kimberley
authors.initials: K
authors.lastname: Evans
authors.affiliation: Wellcome Trust-Medical Research Council Cambridge Stem Cell Institute & Department of Veterinary Medicine, University of Cambridge, Cambridge, CB2 1QR, United Kingdom.
authors.forename: Moritz
authors.initials: M
authors.lastname: Matthey
authors.affiliation: Wellcome Trust-Medical Research Council Cambridge Stem Cell Institute & Department of Veterinary Medicine, University of Cambridge, Cambridge, CB2 1QR, United Kingdom.
authors.forename: Katrin
authors.initials: K
authors.lastname: Volbracht
authors.affiliation: Wellcome Trust-Medical Research Council Cambridge Stem Cell Institute & Department of Veterinary Medicine, University of Cambridge, Cambridge, CB2 1QR, United Kingdom.
authors.forename: Sylvia
authors.initials: S
authors.lastname: Agathou
authors.affiliation: MRC Centre for Developmental Neurobiology, King's College London, London, SE1 1UL, United Kingdom.
authors.forename: Jana
authors.initials: J
authors.lastname: Mukanowa
authors.affiliation: MRC Centre for Developmental Neurobiology, King's College London, London, SE1 1UL, United Kingdom.
authors.forename: Juan
authors.initials: J
authors.lastname: Burrone
authors.affiliation: Wellcome Trust-Medical Research Council Cambridge Stem Cell Institute & Department of Veterinary Medicine, University of Cambridge, Cambridge, CB2 1QR, United Kingdom. rk385@cam.ac.uk.
authors.forename: Ragnhildur T
authors.identifier: http://orcid.org/0000-0001-9675-2722
authors.initials: RT
authors.lastname: Káradóttir
mesh_terms: D000818:Animals; D000975:Antioxidants / Q000032:analysis / Q000494:pharmacology; D002478:Cells, Cultured; D003470:Culture Media / Q000737:chemistry* / Q000494:pharmacology; D005434:Flow Cytometry / Q000379:methods; D006801:Humans; D008027:Light / Q000009:adverse effects*; D009457:Neuroglia / Q000166:cytology / Q000187:drug effects / Q000528:radiation effects*; D009474:Neurons / Q000166:cytology / Q000187:drug effects / Q000528:radiation effects*; D061848:Optical Imaging / Q000379:methods; D051381:Rats
chemical_list: D000975:Antioxidants; D003470:Culture Media