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Cells were plated in triplicate in 35mm plastic cards culture dishes at densities of 1 103 and 0.5 103 Culture dishes were transported at 37 in 100% humidified air with 5% CO2. Colonies comprising of more than 50 cells were scored after incubation for 14 days.Aliquots of 2 108 cryopreserved nucleated units (In 1 cubic centimeters) Were watered down in 9 ml of Xvivo 10 medium (Biowhittaker, Bohringer, australia) And cultured in 25 cm2 tissue culture flasks (Nunc, Oslo, Denmark, Final occurrence, 2 107 person's recombinant IL2 (which is available from Chiron, Ratingen, the netherlands) Was added at the start of the culture at a concentration of 1000 units All flasks were placed in a humidified incubator with 5% CO2 and maintained at 37 for 7 days. via days 2, 4 and as a result 7, Supernatant cells were harvested and adherent cells were detached by scraping with a rubber cop.
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