4 layers instead of a monolayer in cell culture
Posted 19 February 2003 - 08:27 AM
I am fairly new at cell culture, so I hope you will forgive me any stupid question!
I have a primary culture of epithelial cells from the bull epididymis seeded out in Nunc 0.4Ám polycarbonate culture inserts. As I know from electron microscopy, the cells do not stop to divide, when they reach confluency, but rather grow in up to 4 layers over each other. That is bad, because I want to do some transport studies with a monolayer.
Culture medium is RPMI-1640 plus 1mM sodium pyruvate, 100nM Insuline, 200nM hydrocortisone, 200nM testosterone, 1 ÁM dihydrotestosterone and 10% FBS.
Cells growing over each other are definitely epithelial cells, for contamination with other cells is rare.
What can I do to get a monolayer?
Thanks a lot,
Posted 28 April 2003 - 05:57 AM
Posted 09 June 2003 - 12:20 PM