steffen@hawk.bcm.tmc.edu (David Steffen) (08/10/90)
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@mbir.bcm.tmc.edu)