A rapid micro-method for the phytohaemagglutinin-induced human lymphocyte transformation test.

摘要: A quantitative in vitro method for phytohaemagglutinin (PHA) induced lymphocyte transformation is described. This has the following advantages. Cultures require as few 105 lymphocytes and are performed microtrays, over a short incubation period (48 hr). A pulse time of 4 hr with [3H]thymidine utilized. enables high levels to be maintained throughout labelling period, ensures maximum incorporation thymidine into cellular DNA, diminishes damage by internal irradiation. The extraction process been simplified using communal washing procedure after drying cultures on glass-fibre filter discs. The both quicker more reproducible (coefficient variation 6%) than filtration manifolds 23%). The effect adjusting variables such number cells, time, concentration [3H]thymidine, PHA, specific activity [3H]-thymidine duration studied order approach optimal conditions assay system. The reproducibility investigated repeated testing normal individuals day-to-day basis extended periods. mean coefficient samples daily longer intervals (weeks or months between samples) was 15%.