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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-3519 are HOS cells, a human osteosarcoma cell line, expressing CCR5. ARP-3519 are CD4 negative. The parental cell line is available as ARP-3313.
ARP-3519 was generated by subcloning cDNA encoding CCR5 into the retroviral vector pBABE-puro. Amphotropic virus stocks were prepared by cotransfecting 293T cells with the resulting pBABE-puro construct, a VSV-G envelope expression vector, and pSV-gag-pol.?-env-. Supernatants were collected after 48 hours and used to infect HOS CD4+ Cells (ARP-3313). After another 48 hours, cells were selected in medium containing 1 µg per mL puromycin. ß-chemokine receptor expression can be maintained by culturing the cells in medium containing 0.5 µg per mL to 1.0 µg per mL puromycin.
ARP-3519 is also referred to as HOS.CCR5.
Growth Characteristics: Thaw cells quickly at 37°C and immediately place them in 10 mL culture medium. Centrifuge at 400 × g to wash out DMSO, resuspend the cells in 10 mL fresh culture medium, and plate them onto a 10 cm2 cell culture dish. Cells normally require a minimum of 3 to 4 days to recover, but should be checked daily to see if they need to be split. Cells split at 1:10 should become confluent after 3 days. Trypsinize and split at least twice per week; do not allow them to become over confluent.
Growth Medium: DMEM supplemented with 10% fetal bovine serum and 0.5 µg per mL to 1.0 µg per mL puromycin
ARP-3519 is negative for bacteria, fungi and Mycoplasma.
Each vial of ARP-3519 contains frozen cells in DMEM supplemented with 20% fetal bovine serum and 10% dimethyl sulfoxide (DMSO). Please refer to the appropriate data sheet for lot-specific information.
Deng, H., et al. “Identification of a Major Co-Receptor for Primary Isolates of HIV-1.” Nature 381 (1996): 661-666. PubMed: 8649511. Landau, N. L. and D. R. Littman. “Packaging System for Rapid Production of Murine Leukemia Virus Vectors with Variable Tropism.” J. Virol. 66 (1992): 5110-5113. PubMed: 1321291.
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