The life and death of sponge cells
Sipkema, D. ; Snijders, A.P.L. ; Schroën, C.G.P.H. ; Osinga, R. ; Wijffels, R.H. - \ 2004
Biotechnology and Bioengineering 85 (2004)3. - ISSN 0006-3592 - p. 239 - 247.
marine natural-products - in-vitro - growth-inhibition - primary culture - ammonium ion - heat-stress - trehalose - sea - identification - cytotoxicity
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 greater than or equal to22degreesC, but they are insensitive to ammonium concentrations of up to 25 mM. Amphotericin B, which is frequently used in sponge-cell culture media, was found to be toxic to S. domuncula cells. (C) 2004 Wiley Periodicals, Inc.