Hi, This message is related to the CHO cell sorting thread. I perform many sorts with HEK 293 and CHO transfected with different receptors. I generally perform single cell deposition into each well of 96 well plates to establish clonal cell lines. My problem is getting these single cell clones to grow especially with the HEK 293 (I have much better success generating CHO lines). I have tried all sorts of little tricks to get these cells to grow such as using conditioned media in the wells, and using collagen- or poly-D-lysine-coated plates. My success rate with single cell deposition with HEK 293 is quite poor, I am estimating its probably below 1%. Meaning, I am lucky to generate one clone per 96 well plate. Of course, success depends quite a bit on the gene transfected into the HEK 293 and the health of the cells at the time of sorting. Does anybody know of any conditions that would increase the number of clones I could generate using HEK 293? Thanks, Rich Richard C. Hastings Associate Scientist Target Expression/LDD 1800 Concord Pike Wilmington, DE 19803 302-886-2452 richard.hastings@astrazeneca.com
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