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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-2091 is a culture of HLM30 cells that are CD4+ and negative for virus particle production, but can be stimulated to produce non-infectious virions. ARP-2091 was generated by transducing HeLa CD4+ cells (ARP-154, contributed by Dr. Richard Axel) with the tat-defective mutant pMtat30 (ARP-2087), which contains a TGT→GGT substitution at cysteine residue 30 in the tat reading frame . The HIV proviral DNA was derived from pHXB2gpt, an infectious molecular clone of HIV-1 IIIB (ARP-398, contributed by Dr. R. Gallo).
HLM30 cells are negative for virus particle production, but will produce HIV-1 after cocultivation with tat-expressing cells, or after exposure to UV light and subsequent coculture with H9 cells. The resultant virus is a mixture of infectious revertants and defective HIV-1 mutants.
Growth Characteristics: These cells can be maintained in culture by adding fresh medium to the adherent cells every 3 to 4 days. Trypsinization is necessary only once every month or so unless the cells become over-confluent or need to be transferred to an additional flask. These cells can be adapted for growth in richer media and in fetal bovine serum; however, maintaining them in MEM with 5% horse serum is less likely to cause the cells to spontaneously produce background virus.
Growth Medium: Minimal Essential Medium (MEM) supplemented with 5% horse serum and 10 µg per mL gentamicin
ARP-2091 is negative for bacteria, Mycoplasma and fungi. Each vial of ARP-2091 contains approximately 6 × 106 cells in MEM supplemented with 20% horse serum and 10% dimethyl sulfoxide (DMSO). Please refer to the attached file(s) for more information.
Sadaie, M. R., et al. “Activation oftat-Defective Human Immunodeficiency Virus by Ultraviolet Light.” New Biol. 2 (1990): 479-486. PubMed: 1981148.
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