The lines are based on the 293T cell line (a human embryonic kidney line transformed with adenovirus E1a and carrying a temperature sensitive T antigen co-selected with neomycin). The unique feature of this cell line is that it is highly transfectable with either calcium phosphate mediated transfection or lipid-based transfection protocols-- up to 50% or higher of cells can be transiently transfected. Chloroquine rheumatoid arthritis mechanism of action Plaquenil and pepto bismol Chloroquine fisher A. Thawing Phoenix™ Cells 1. Remove the vial containing frozen cells from liquid nitrogen or shipping box. Thaw rapidly at 37°C by hold-ing the vial and gently shaking in the water bath. Take out the vial from the water bath when the frozen cells start to thaw about 1-2 minutes. The key point is NOT to let the cells thaw completely. 2. A severe eye problem has happened with chloroquine. This may lead to lasting eyesight problems. The risk may be higher if you have some types of eye or kidney problems. The risk may also be higher with some doses of chloroquine, if you use chloroquine for longer than 5 years, or if you take certain other drugs like tamoxifen. Phoenix is a second-generation retrovirus producer cell line for the generation of helper-free ecotropic and amphotropic retroviruses. The unique feature of this cell line is that it is highly transfectable with either calcium phosphate mediated transfection or lipid-based transfection protocols-- up to 50% or higher of cells can be transiently transfected. The lines offered advantages over previous stable systems in that virus can be produced in just a few days. The lines were created by placing into 293T cells constructs capable of producing gag-pol, and envelope protein for ecotropic and amphotropic viruses. Chloroquine phoenix cells Retrovirus preparation using Phoenix cells, Chloroquine Indications, Side Effects, Warnings - Months for hydroxychloroquine to work on joints Seed 293T packaging cells at 3.8×10 6 cells per plate in DMEM complete in 10 cm tissue culture plates. Incubate the cells at 37 ℃, 5% CO 2 for ~20 hours. Gently aspirate media, add 10 mL fresh DMEM complete containing 25 μM cloroquine diphosphate and incubate ~5 hours. For 10 mL of DMEM complete, add 10 μL of 25 mM chloroquine diphosphate. Addgene Lentivirus Production Protocol. Phoenix-AMPHO ATCC ® CRL-3213™ Homo sapiens kidney. Can the phoenix cell be transfected using lipofectamine.. Phoenix is a second-generation retrovirus producer lines for the generation of helper free ecotropic and amphotropic retroviruses. The lines are based on the 293T cell line a human embryonic kidney line transformed with adenovirus E1a and carrying a temperature sensitive T antigen co-selected with neomycin. Chloroquine-treated ARPE-19 cells demonstrate a marked increase in vacuolation and dense intracellular debris. These are identified as chloroquine-dilated lysosomes and lipid bodies with LAMP-2 and LipidTOX co-localization, respectively. Dilation is an indicator of lysosomal dysfunction. Chloroquine is a 4-aminoquinoline with antimalarial, anti-inflammatory, and potential chemosensitization and radiosensitization activities. Although the mechanism is not well understood, chloroquine is shown to inhibit the parasitic enzyme heme polymerase that converts the toxic heme into non-toxic hemazoin, thereby resulting in the accumulation of toxic heme within the parasite.