The life and death of sponge cells.
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 >or=22 degrees C, 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.
Sipkema
Detmer
D
Food and Bioprocess Engineering Group, Wageningen University, PO Box 8129, 6700 EV Wageningen, The Netherlands. detmer.sipkema@wur.nl
Snijders
Ambrosius P L
AP
Schroën
Carin G P H
CG
Osinga
Ronald
R
Wijffels
René H
RH
D000666:Amphotericin B / Q000494:pharmacology*; D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects / Q000502:physiology*; D002470:Cell Survival / Q000187:drug effects / Q000502:physiology; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D005434:Flow Cytometry / Q000379:methods*; D011161:Porifera / Q000166:cytology* / Q000187:drug effects; D000644:Quaternary Ammonium Compounds / Q000494:pharmacology*; D013050:Spectrometry, Fluorescence / Q000379:methods; D013696:Temperature
D000900:Anti-Bacterial Agents; D000644:Quaternary Ammonium Compounds; D000666:Amphotericin B