Journal of Immunological Methods 1987-12-04

A fluorometric assay for determining cell growth in lymphocyte proliferation and lymphokine assays.

E N Dotsika, C J Sanderson, Eleni N. Dotsika, Colin J. Sanderson

Index: J. Immunol. Methods 105(1) , 55-62, (1987)

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Abstract

A microplate method for assessing cell growth and viability based on the hydrolysis of fluorogenic substrates by cell esterases has been investigated. Living cells incubated with fluorescein diacetate or 4-methylumbelliferyl heptanoate generate a fluorescent product which is proportional to the number of cells. This can be used as a simple and economical readout for various bioassays such as, for example, the assessment of IL-2 and IL-3 on factor-dependent cell lines, and on antigen- and mitogen-stimulated proliferation of lymphocytes. This fluorometric assay has similar sensitivity to the measurement of [3H]thymidine uptake and greater sensitivity than standard colorimetric assays. Incubation with MUH for a period of 30-60 min at 22 degrees C is adequate.