A new methodology for plant cell viability assessment using intracellular esterase activity

A plant cell suspension culture of Alfalfa (Medicago sativa L.) was grown in a bioreactor using a batch procedure. The cytoplasmic esterase activity (EC 3.1) was extracted from the cells and measured during cultivation using fluorescein diacetate as the fluorogenic substrate. This enzymatic activity...

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Bibliographic Details
Published inPlant cell reports Vol. 19; no. 2; pp. 171 - 176
Main Authors Steward, N, Martin, R, Engasser, J M, Goergen, J L
Format Journal Article
LanguageEnglish
Published Germany Springer Nature B.V 01.12.1999
Subjects
Alfalfa
Bioreactors
Cells
Enzymatic activity
Microbiology
Viability
Cytoplasmic esterase activity
Key words Plant cell culture
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Summary:A plant cell suspension culture of Alfalfa (Medicago sativa L.) was grown in a bioreactor using a batch procedure. The cytoplasmic esterase activity (EC 3.1) was extracted from the cells and measured during cultivation using fluorescein diacetate as the fluorogenic substrate. This enzymatic activity was conclusively found to be correlated to cell viability assessed with the membrane integrity test using the trypan blue dye. This new viability determination method is convenient, simple and can be reproduced because: (1) the difficult step of counting the cells when using the trypan blue exclusion method is avoided and (2) the esterase activity level per viable cell constituted of numerous enzymes depends on cell viability but is independent of cellular metabolism.
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ISSN:0721-7714
1432-203X
DOI:10.1007/s002990050729