Measurement of cell death by oxidative stress in three-dimensional spheroids from trophoblast and in fragments of decidua tissue.
We report a new morphometric method for measurement of the amount of cell death in three-dimensional multicellular spheroids of the trophoblast-like cell line AC1-M59 and of cultured pieces of decidua tissue (decidua spheroids) in response to a cytotoxic agent. The viability of the spheroids was assessed by adding propidium iodide to the culture medium at the end of the toxic treatment. On fluorescence and brightfield images of serial cryosections the areas of propidium iodide fluorescence and the entire corresponding spheroids were measured by applying digital image processing and ratiometrical quantification. As an example, we evaluated the cytotoxic effect of hydrogen peroxide on both types of spheroids. The relative potency of hydrogen peroxide to induce tissue damage was assessed quantitatively for determination of the minimal concentration that leads to an increase in cytotoxicity. The method presented suggests general applicability for in vitro determination of toxicity against tissues.
Institut für Zellbiologie, Histologie und Embryologie, Medizinische Universität Graz, Harrachgasse 21, A-8010 Graz, Austria.
Regine-Susanne
RS
Theuerkauf
Helmut
H
Ahammer
Monika
M
Siwetz
Christine
C
Helige
Gottfried
G
Dohr
Wolfgang
W
Walcher
José Ramón
JR
Palacio
Paz
P
Martinez
Peter
P
Sedlmayr
D016923:Cell Death / Q000187:drug effects*; D045744:Cell Line, Tumor; D019411:Clinical Laboratory Techniques; D003656:Decidua / Q000187:drug effects / Q000473:pathology*; D005260:Female; D006801:Humans; D006861:Hydrogen Peroxide / Q000633:toxicity; D007091:Image Processing, Computer-Assisted; D007202:Indicators and Reagents / Q000494:pharmacology; D008856:Microscopy, Fluorescence; D016877:Oxidants / Q000633:toxicity; D018384:Oxidative Stress / Q000502:physiology*; D011419:Propidium / Q000494:pharmacology; D018874:Spheroids, Cellular / Q000187:drug effects / Q000473:pathology*; D014327:Trophoblasts / Q000187:drug effects / Q000473:pathology*
D007202:Indicators and Reagents; D016877:Oxidants; D011419:Propidium; D006861:Hydrogen Peroxide