Materials and Reagents

1. CD4⁺ T cell line H9 cells (ATCC HTB-176 )
   
2. Fetal bovine serum (FBS) (Thermo Fisher Scientific, catalog number: SH30071.03 HI )
   Note: This particular FBS has been tested by the author, but may be substituted with FBS from different suppliers as desired by users.
   
3. Penicillin-Streptomycin liquid (Life Technologies, Gibco^®, catalog number: 15070-063 )
   
4. Cell culture media: RPMI1640 (Life Technologies, Gibco^®, catalog number: 11875-093 ) (see Recipes)
   

Equipment

1. Bench-top centrifuges
   
2. CO₂ incubator
   
3. -80 °C freezer
   
4. 50 ml conical tubes (BD Biosciences, Falcon^®, catalog number: 35-2070 )
   
5. 5, 10, 25-ml pipet (BD Biosciences, Falcon^®, catalog number: 35-7501 , 35-7554 , 35-7556 )
   
6. T-75, T-175 cell culture flask (BD Biosciences, Falcon^®, catalog number: 35-3136 , 35-3112 )
   
7. CryoTube Vials (NUNC, 377267)
   

Procedure