We have been growing primary human sinonasal epithelial cells in trans-wells at p2, until confluence. They are grown to confluence in BEGM (Lonza), and switched to a 50:50 mixture of BEGM and DMEM with an addition of Retinol placed in the basolateral side of the trans-well. Theoretically, these cultures should be able to last for about 6 months, but we have experienced considerable leaching about a month and a half into the ALI. We tried making a new batch of Retinol, but are still experiencing leaching. Has anyone else struggled with this, and if so what did you do about it?
Edited by wendy245, 10 October 2012 - 09:00 AM.