1. Cells undergoing first passage should form confluent monolayers 5-10 d from initial seeding.

2. The media should be changed at 24 h after which time small islands of attached cells are visible under phase-contrast microscopy.

3. The media should be exchanged every 48 h thereafter, until the monolayers are 50-60% confluent. Then the media should be changed every 24 h.

4. Once confluent, the cells can be trypsinised and passaged or cryopreserved.


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.