A simple method for classification of cell death by use of thin layer collagen gel for the detection of apoptosis and/or necrosis after cancer chemotherapy
A. Matsuo et al., A simple method for classification of cell death by use of thin layer collagen gel for the detection of apoptosis and/or necrosis after cancer chemotherapy, JPN J CANC, 92(7), 2001, pp. 813-819
To assess the efficacy of cancer chemotherapy, an important index is apopto
sis of the target cells, which can usually be confirmed by electron microsc
opy (EM). We established a new experimental technique, whereby cancer cells
(MKN45) were distributed in thin collagen gel as one or two cell layers, a
nd cultured with anti-cancer drugs (5-FU and CDDP). The cells were stained
with fluorescent Hoechst 33258 (Ho) and photographed, then with hematoxylin
and eosin (H&E) and again photographed, and processed for EM. This approac
h allowed us to characterize the patterns of death of single cells in detai
l. There were six patterns of cell damage: two patterns of apoptosis, early
peripheral condensation of chromatin and late apoptotic bodies, two patter
ns of necrosis, cytoplasmic swelling and washed-out images, and two further
patterns, with morphological features of both apoptosis and necrosis, neit
her classified into necrosis nor apoptosis. The results show that cell deat
h patterns can be mostly determined by combining observations of Ho and H&E
-stained cells without the necessity for EM observation.