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The life and death of sponge cells
Author(s) -
Sipkema Detmer,
Snijders Ambrosius P. L.,
Schroën Carin G. P. H.,
Osinga Ronald,
Wijffels René H.
Publication year - 2003
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.10886
Subject(s) - sponge , propidium iodide , viability assay , biology , microbiology and biotechnology , cell culture , fluorescence microscope , flow cytometry , cell , biochemistry , fluorescence , programmed cell death , botany , apoptosis , genetics , physics , quantum mechanics
Cell viability is an essential touchstone in the study of the effect of medium components on cell physiology. We developed a flow‐cytometric assay to determine sponge‐cell viability, based on the combined use of fluorescein diacetate (FDA) and propidium iodide (PI). Cell fluorescence measurements based on incubation of cells with FDA or PI resulted in a useful and reproducible estimate of the viability of primary sponge‐cell cultures. We studied the effects of temperature, ammonium, and the fungicide amphotericin B on the viability of a primary‐cell culture from the marine sponge Suberites domuncula using the aforementioned flow‐cytometric assay. S. domuncula cells die rapidly at a temperature of ≥22°C, but they are insensitive to ammonium concentrations of up to 25 m M . Amphotericin B, which is frequently used in sponge‐cell culture media, was found to be toxic to S. domuncula cells. © 2004 Wiley Periodicals, Inc.