Lactate dehydrogenase assay for assessment of polycation cytotoxicity.
Cellular toxicity and/or cell death entail complex mechanisms that require detailed evaluation for proper characterization. A detailed mechanistic assessment of cytotoxicity is essential for design and construction of more effective polycations for nucleic acid delivery. A single toxicity assay cannot stand alone in determining the type and extent of damage or cell death mechanism. In this chapter we describe a lactate dehydrogenase (LDH) assay for high-throughput screening that can be used as a starting point for further detailed cytotoxicity determination. LDH release is considered an early event in necrosis but a late event in apoptosis. An accurate temporal assessment of the toxic responses is crucial as late apoptosis may convert into necrosis as well as in situations where cell death is initiated without any visible cell morphological changes or responses in assays measuring late events, resulting in early ongoing toxicity being overlooked.
Centre for Pharmaceutical Nanotechnology, University of Copenhagen, Copenhagen Ø, Denmark.
Ladan
L
Parhamifar
Helene
H
Andersen
S Moein
SM
Moghimi
D002452:Cell Count; D016923:Cell Death / Q000187:drug effects; D002460:Cell Line; D003470:Culture Media / Q000378:metabolism; D003603:Cytotoxins / Q000633:toxicity*; D057075:Enzyme Assays / Q000379:methods*; D007770:L-Lactate Dehydrogenase / Q000378:metabolism*; D011073:Polyamines / Q000633:toxicity*; D000071228:Polyelectrolytes
D003470:Culture Media; D003603:Cytotoxins; D007770:L-Lactate Dehydrogenase; D011073:Polyamines; D000071228:Polyelectrolytes