Viability measurements of hybridoma cells in suspension cultures |
| |
Authors: | José M. Coco-Martin Jan W. Oberink Tiny A. M. van der Velden-de Groot E. Coen Beuvery |
| |
Affiliation: | (1) National Institute of Public Health and Environmental Protection, P. O. Box 1, 3720 BA Bilthoven, The Netherlands;(2) Division of Experimental Therap, The Netherlands Cancer Institute, Plesmanlaan 121, 1066 CX Amsterdam, The Netherlands |
| |
Abstract: | Several methods were applied to determine the viability of hybridoma cells in suspension. These methods include dye inclusion and exclusion assays such as the classical trypan blue exclusion assay, the propidium iodide (PI) exclusion assay and the fluorescein diacetate (FDA) inclusion assay. Furthermore, the relation was studied between release of lactate dehydrogenase (LDH) by hybridoma cells and their viability. Also the ATP content of the cells and cellular heterogeneity as measured with a flow cytometer were determined in relation to cellular viability. The dye inclusion and exclusion assays using trypan blue, FDA, PI were shown to be useful methods to determine cellular viability. With the FDA and PI methods it was possible to obtain additional information about cells which are in a transition state between viable and non-viable. The viability according to the scatter properties of the cells appears to reflect the overall condition of the cells, although interpretation of the results is difficult. Measurement of LDH release in the culture fluid or the cytoplasmic ATP content could not be used as parameters for cell viability. |
| |
Keywords: | viability hybridoma fluorescein diacetate propidium-iodide flow cytometry |
本文献已被 SpringerLink 等数据库收录! |
|