A fluorometric assay for determining cell growth in lymphocyte proliferation and lymphokine assays.
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.
Dotsika
E N
EN
National Institute for Medical Research, Mill Hill, London, U.K.
Sanderson
C J
CJ
D001681:Biological Assay / Q000379:methods; D002460:Cell Line; D004950:Esterases / Q000378:metabolism; D019793:Fluorescein; D005452:Fluoresceins; D005456:Fluorescent Dyes; D006923:Hymecromone / Q000031:analogs & derivatives*; D007158:Immunologic Techniques; D007376:Interleukin-2 / Q000032:analysis; D007377:Interleukin-3 / Q000032:analysis; D007700:Kinetics; D008213:Lymphocyte Activation; D008222:Lymphokines / Q000032:analysis*; D013050:Spectrometry, Fluorescence; D013329:Structure-Activity Relationship; D014468:Umbelliferones
D005452:Fluoresceins; D005456:Fluorescent Dyes; D007376:Interleukin-2; D007377:Interleukin-3; D008222:Lymphokines; D014468:Umbelliferones; C054268:4-methylumbelliferyl heptanoate; D006923:Hymecromone; D004950:Esterases; D019793:Fluorescein