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

Citation
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
Citations number
27
Categorie Soggetti
Onconogenesis & Cancer Research
Journal title
JAPANESE JOURNAL OF CANCER RESEARCH
ISSN journal
09105050 → ACNP
Volume
92
Issue
7
Year of publication
2001
Pages
813 - 819
Database
ISI
SICI code
0910-5050(200107)92:7<813:ASMFCO>2.0.ZU;2-J
Abstract
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.