Notes on Technic: A simple procedure for autoradiography of keratinocyte cultures

Abstract

Although cell cultures usually grow as a monolayer, there are some circumstances in which they become multilayered or stratified. This is the case with epidermal cell cultures where skin keratinocytes grow attached to the bottom of the culture flask, but as they differentiate lose contact with the bottom and pile up on top of the cell colony to create a situation resembling the structure of skin in vivo (Rheinwald 1980). In this way, proliferative cell pools are attached to the bottom of the culture flask and are partly covered by nonproliferative, differentiated cells. This particular arrangement makes cell kinetic studies difficult because such studies are usually carried out by autoradiographic evaluation of the uptake of tritium-labeled DNA precursors (mainly thymidine) into the nucleus of cells synthesizing DNA.