Steward N, Martin R, Engasser J M, Goergen J L
Laboratoire des Sciences du Génie Chimique, Institut National Polytechnique de Lorraine - CNRS, BP 172, F-54505 Vandoeuvre-lès Nancy, France e-mail:
Plant Cell Rep. 1999 Dec;19(2):171-176. doi: 10.1007/s002990050729.
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.
采用分批培养法在生物反应器中培养紫花苜蓿（Medicago sativa L.）的植物细胞悬浮培养物。从细胞中提取细胞质酯酶活性（EC 3.1），并在培养过程中使用荧光素二乙酸作为荧光底物进行测量。通过使用台盼蓝染料进行膜完整性测试评估细胞活力，最终发现这种酶活性与细胞活力相关。这种新的活力测定方法方便、简单且可重复，原因如下：（1）避免了使用台盼蓝排除法时计数细胞这一困难步骤；（2）由多种酶组成的每个活细胞的酯酶活性水平取决于细胞活力，但与细胞代谢无关。
