title: Experimental survey of non-clonogenic viability assays for adherent cells in vitro.
abstract: Results of rapid cell viability assays were experimentally compared in order to reveal the most suitable test for in vitro investigations of the combination of photodynamic therapy (PDT) with chemotherapeutic drugs. meso-Tetra(3-hydroxyphenyl)-chlorin (m-THPC) accumulating in cell membranes and meso-tetra(4-sulfonatophenyl)-porphin (TPPS4) accumulating in lysosomes were used as photosensitisers. Doxorubicin that localises, mainly, to nucleus and vincristine that binds to microtubules were used as cytostatic drugs. Two adherent rodent cell lines, baby hamster kidney (BHK-21) and murine hepatoma (MH-22A), were used to examine the contribution of a cell. We tested cytotoxicity assays of the main groups of fast (non-clonogenic) methods of cell viability measuring. Plasma membrane integrity was estimated by trypan blue exclusion and LDH leakage, metabolic activity was tested by [3H]-thymidine incorporation and MTT assay, loss of monolayer adherence was measured by staining with crystal violet and CyQUANT. The most sensitive test in each case was the assay related to the site of the direct damage, and measurement of the loss of monolayer adherence proved to be as sensitive assay as the damage-specific one. All the assays applied, except for the LDH release, revealed a higher effect of combination of m-THPC-mediated phototreatment and doxorubicin compared to either of the single treatments.
authors.affiliation: Department of Biochemistry and Biophysics, Vilnius University, Ciurlionio 21, LT-03101 Vilnius, Lithuania.
authors.forename: Ingrida
authors.initials: I
authors.lastname: Mickuviene
authors.forename: Vida
authors.initials: V
authors.lastname: Kirveliene
authors.forename: Benediktas
authors.initials: B
authors.lastname: Juodka
mesh_terms: D000818:Animals; D001681:Biological Assay; D002448:Cell Adhesion / Q000187:drug effects; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D002470:Cell Survival / Q000187:drug effects*; D002999:Clone Cells; D006224:Cricetinae; D004317:Doxorubicin / Q000494:pharmacology*; D004338:Drug Combinations; D005562:Formazans / Q000378:metabolism; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008027:Light; D008652:Mesoporphyrins / Q000494:pharmacology*; D051379:Mice; D017319:Photosensitizing Agents / Q000494:pharmacology*; D011166:Porphyrins / Q000494:pharmacology*; D013778:Tetrazolium Salts / Q000378:metabolism; D013936:Thymidine / Q000378:metabolism
chemical_list: D004338:Drug Combinations; D005562:Formazans; D008652:Mesoporphyrins; D017319:Photosensitizing Agents; D011166:Porphyrins; D013778:Tetrazolium Salts; C079782:MTT formazan; C016136:tetraphenylporphine sulfonate; D004317:Doxorubicin; D007770:L-Lactate Dehydrogenase; C072269:temoporfin; D013936:Thymidine