Isolating clones of HL60 cells

David Steffen steffen at hawk.bcm.tmc.edu
Fri Aug 10 09:43:30 EST 1990


  Has anyone had any luck cloning HL60 cells?  We are trying to isolate stably 
transfected HL60 cells, and are having good luck transfecting the cells by 
electroporation (assayed by transient transfection) but are unable to isolate 
single cell-derived clones by selection after transfection.  We have been 
unsuccessful even if we just try to isolate clones without transfection or 
selection.

Things we have tried:
--------------------
  We have tried isolating clones either as colonies in soft agar or by limiting 
dilution in 96 well plates.  We have tried using conditioned medium.  Any other 
suggestions?  (We have been successful isolating single cell clones of either 
adherent or non-adherent cells from other lines using these techniques.)

  I suggest that people reply to me by email, and I will post a summary of any 
responses to this newsgroup.  Even if I receive no response, I will summarize 
that fact (after about two weeks).  Obviously, responses directly to this 
newsgroup are infinitely preferable to no response.

  Thanks in advance!

-David Steffen- (steffen at mbir.bcm.tmc.edu)



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