title: Evaluation of a continuous quantification method of apoptosis and necrosis in tissue cultures.
abstract: In tissue-engineering and other life sciences, there is a growing need for real-time, non-destructive information on apoptosis and necrosis in 2D and 3D tissue cultures. Previously, propidium iodide was applied as a fluorescent marker for monitoring necrosis. In the current study this technique was extended with a fluorescent apoptosis marker, YO-PRO-1, to discriminate between both stages of cell death. The main goal was to evaluate the performance of YO-PRO-1 and propidium iodide during monitoring periods of up to 3 days. Apoptosis was induced in C2C12 cultures and the numbers of YP-positive and PI-positive nuclei were counted in time. The performance of the dual staining was evaluated with a metabolic measure and a probe intensity study. Cell metabolism was unaffected during the first 24 h of testing. In conclusion, the YP/PI dual staining method was found to be a powerful tool in obtaining real-time spatial information on viability in cell and tissue culture without culture disruption.
authors.affiliation: Department of Biomedical Engineering, Eindhoven University of Technology, WH 4.105, P.O. Box 513, 5600 MB, Eindhoven, The Netherlands, d.gawlitta@tue.nl.
authors.forename: Debby
authors.initials: D
authors.lastname: Gawlitta
authors.forename: Cees W J
authors.initials: CW
authors.lastname: Oomens
authors.forename: Frank P T
authors.initials: FP
authors.lastname: Baaijens
authors.forename: Carlijn V C
authors.initials: CV
authors.lastname: Bouten