Comparison of double fluorescence staining and LDH-test for monitoring cell viability in vitro.
To examine glutamate-mediated toxic effects in dispersed cultures of the rat cerebral cortex we compared the utility of parameters of cell viability in parallel. A 1 h exposure to glutamate and glutamate analogues (kainate, quinolinate, NMDA) was found to produce typical morphological changes in matured cell cultures. The double-labelling fluorescence technique (fluorescein diacetate/propidium iodide) reflected the degeneration process vividly. A significant increase in LDH-activity released into the medium was noted only when the cells were stressed but still living. When the phase of progressive cell death was running, LDH activity in the medium decreased markedly. Obviously, increasing LDH release in the culture medium must be considered rather to be an indicator for a slowly evolving degenerative process than for cell death.
Keilhoff
G
G
Institute of Medical Neurobiology, Medical Academy of Magdeburg, Germany.
Wolf
G
G
D000818:Animals; D015415:Biomarkers / Q000032:analysis; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000166:cytology / Q000187:drug effects* / Q000196:embryology; D017077:Culture Media, Conditioned / Q000737:chemistry; D003584:Cytological Techniques; D005452:Fluoresceins / Q000378:metabolism; D005456:Fluorescent Dyes / Q000378:metabolism; D005971:Glutamates / Q000633:toxicity*; D018698:Glutamic Acid; D006868:Hydrolysis; D007608:Kainic Acid / Q000633:toxicity; D007770:L-Lactate Dehydrogenase / Q000032:analysis*; D008856:Microscopy, Fluorescence; D016202:N-Methylaspartate / Q000633:toxicity; D009410:Nerve Degeneration / Q000187:drug effects; D009419:Nerve Tissue Proteins / Q000032:analysis*; D009457:Neuroglia / Q000187:drug effects* / Q000378:metabolism; D009474:Neurons / Q000187:drug effects* / Q000378:metabolism; D009498:Neurotoxins / Q000633:toxicity*; D011419:Propidium / Q000378:metabolism; D017378:Quinolinic Acid / Q000633:toxicity; D051381:Rats; D017208:Rats, Wistar
D015415:Biomarkers; D017077:Culture Media, Conditioned; D005452:Fluoresceins; D005456:Fluorescent Dyes; D005971:Glutamates; D009419:Nerve Tissue Proteins; D009498:Neurotoxins; D011419:Propidium; D018698:Glutamic Acid; D016202:N-Methylaspartate; D007770:L-Lactate Dehydrogenase; D017378:Quinolinic Acid; D007608:Kainic Acid; C018506:diacetylfluorescein