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Are normal or immortalized human epithelial cells clonogenic?

Tissue culture Isogenic clonogenic Human epithelial cells

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#1 5280

5280

    Veteran

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Good

Posted 08 July 2015 - 05:17 AM

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!

 

 



#2 bob1

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    Thelymitra pulchella

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Excellent

Posted 08 July 2015 - 12:33 PM

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.







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