A new methodology for plant cell viability assessment using intracellular esterase activity |
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Authors: | N Steward R Martin J M Engasser J L Goergen |
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Institution: | (1) Laboratoire des Sciences du Génie Chimique, Institut National Polytechnique de Lorraine – CNRS, BP 172, F-54505 Vandoeuvre-lès Nancy, France e-mail: Jean-Louis.Goergen@ensaia.inpl-nancy.fr Fax: +33-383-59-58-04, FR |
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Abstract: | 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.
Received: 28 January 1999 / Accepted: 1 April 1999 |
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Keywords: | Plant cell culture Viability Cytoplasmic esterase activity |
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