I am having a problem getting H295R cells to produce and secrete aldosterone I am testing the aldosterone level via ELISA method. I have tried both KCL and AngII however, nothing seems to make the aldosterone level rise. I was thinking maybe something is wrong with my cells? They seem to grow really slowly ( I have been using the culture method recommended by ATCC) and seeing one article *Rainey 1993 subcultured the adhesive cells for 3 months before performing experiments? Or maybe the cells have to be at a certain confluent to secrete aldosterone? Any advice would help.