1. As per subculture procedure steps 1-4.

2. Resuspend the cell pellet in complete growth medium containg 10% DMSO, and pipette 1 mL of the cell suspension into each cryovial.

3. Place cryovials into a styrofoam container (to slow freeze), and place container in -70°C for 1 h. Or place cryovials in Nalgene 1°C freezing container with isopropanol in -80°C freezer for 24 h.

4. Transfer vials to liquid nitrogen.


Note: This protocol was taken from "Primary Culture of Human Proximal Renal Tubular Epithelial Cells" by Paul A. Glynne. From: Methods in Molecular Medicine, Vol.36: Septic Shock. Humana Press Inc., Totowa, NJ.