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-100°C or colder, preferably in the vapor phase of a liquid nitrogen freezer
Quantity limit per order for this item is 1. This item can be ordered twice a year. Orders over this limit will be sent to NIAID for approval before shipment. ARP-166 is a hypoxanthine-aminopterin-thymidine (HAT)-sensitive derivative of CEM, a human T-cell line derived from the peripheral blood buffy coat of a four-year-old Caucasian female with acute lymphoblastic leukemia. Strain A3.01 was selected by growth in hypoxanthine and aminopterin-containing medium. It is suitable for human T-lymphocyte fusions. Cells are Leu-3+, Leu-8+, Leu-1+,tac-, transferrin receptor+, sensitive to infection with lymphadenopathy associated virus (LAV), and susceptible to cytopathic effects when infected. Growth Characteristics: When thawing ARP-166, DMSO should be gently washed out with 37°C medium and the initial culture should be seeded at 1 x 106 cells/mL. Cells quickly recover viability over 3 days. Cells should be passaged every three days to give a concentration of 1 x 106 cells/mL. Cells grow in single-cell suspension with a mature lymphocyte morphology and have a doubling time of 24 hours. A3.01 has also been grown successfully in OPTI-MEM® Reduced Sera Medium containing 2.5% fetal bovine serum, 2.0 mM L-glutamine, 100 U/mL penicillin and 100 μg/mL streptomycin. Propagation Medium: The recommended propagation medium is 90% RPMI 1640, 10% heat-inactivated fetal bovine serum and 2.0 mM L-glutamine. Tests for bacteria, fungi and mycoplasma were negative. Each vial of ARP-166 contains approximately 3.52 × 106 cells in 0.8 mL of freeze medium. Post-thaw viability was 52%.
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