Oral Combinational Antiretroviral Therapy to Treat HIV-1 Infection in a Humanized Mouse Model
Transcript
Begin by weighing individual drugs. Make 10 food cups with cART using sterile cell scrapers to weigh out 250 milligrams of FTC, 375 milligrams of TDF, and 500 milligrams of RAL or ELV into individual sterile 50-milliliter centrifuge tubes in a biosafety cabinet. Add DMSO to the FTC and TDF tube to make up a final concentration of 250 milligrams per milliliter, and into the ERL or ELV tube to a final concentration of 500 milligrams per milliliter.
Mix the drug solution by stirring and pipetting until a clear solution is obtained. Sterilize the solutions using a 0.22-micrometer hydrophilic PVDF membrane filter with a sterile syringe. Individual drug solutions can be stored at minus 20 degrees Celsius for 12 weeks.
Freshly thaw one aliquot of each drug solution before use at 37 degrees Celsius until the solution becomes clear. Mix well using a pipette. Combine the drugs dissolved in DMSO, and mix well to make up a master mix containing one milliliter of FTC, 1.5 milliliters of TDF, and 1 milliliter of ELV or RAL.
Add 350 microliters of cART master mixed solution into a to prepare one diet gel boost cART cup. Add 0.75 milliliters of trimethoprim-sulfamethoxazole into the cup. Stir thoroughly using 1-milliliter sterile pipette tips. Using a pipette tip, aliquot the food containing cART onto a 60-millimeter Petri dish.
Calculate the amount of cART in the food cup by weighing the food for each cage according to the number of mice. Remove regular chow from the cage, and replace it with a food cup containing cART. Refresh cART food three times per week. Weigh used cups to monitor intake. Then weigh the mice weekly to confirm consumption.
