I'm hoping to do a clonal expansion of Jurkat, expressing my construct which is currently transduced virally. From the FACS data I currently have roughly 30-40% expression in my culture but I need it to be close to 100% as the effect I'm looking for I believe to get diluted by the reduced presence of my molecule of interest in the total culture. I can do cell sorting but usually what happens is that the 5% cell population that do not express the construct eventually take over after a few passages.
I'm planning to do a 96 well plate format single cell seeding and expansion of Jurkats but it has been done before by another member of our lab and the person suggests that single cell Jurkats (unlike CHO cells) do not survive a single cell state although they are suspension cells and do not necessarily require contact to survive although they clump. Hence I'm wondering if anyone has any protocols they could suggest such as contents of the media (e.g. conditioned media) or any other recommendations for Jurkat single cell clonal expansion.
P.S. I am fully aware that there might not be a single right answer to this and the answer requires a degree of experience/opinion on the working method but I would fully appreciate any helpful suggestions/responses.