Are normal or immortalized human epithelial cells clonogenic? - (Jul/08/2015 )
How do people establish isogenic clones from normal or immortalized human epithelial cells (non-transformed) when the cells are not clonogenic? I am referring to the ability of cells to form clones from single cells on plastic. I am not referring to cells being able to form colonies in soft agar which is a feature of transformed cells.
I want to introduce an inducible transgene into hTERT/p53 R175H immortalized cells then isolate isogenic clones but the cells will not grow as single cells. Will a richer media allow growth from single cells (ie add insulin or other growth factors)?Or do most people just work with polyclonal populations?
Thanks!
You can condition medium (grow cells in the medium for 24-48h, remove medium, filter, then store at -80 if needed), so that it contains the growth factors that cells release while growing as a population, then use this medium to help your clonogenic cells to get going.