I am interested in studying the role of a certain gene in the expression of Collagen type II and chondrogenesis per se in ATDC5 cells. But as per the papers for proper chondrogenesis to happen it takes approximately 10 days. Whereas we transfect the cells probably 2 days after seeding around 60-70% confluency. I am interested to know whether I can transfect at Day 2 and then replace the media with chondrogenic media and allow chondrogenesis to happen for the next few days and expect my transfected cells to have their effect sustained for these many days.
Hi Ankita,
What is the purpose of the transfection? I assume either for silencing or over-expressing a gene. If that the case, you may try stable transfection methods instead of a transient transfection. Then, you can investigate your gene of interest for longer time.
Ahmed E. Dhamad Thank you! Well I am planning to make a stable cell line in case I get some prelimnary data. Apart from that I am planning to transfect with a Tol2 based vector which is a transposon mediated vector system which allows integration into the genome. But as I have seen several papers in which people have done a transient transfection and then gone for chondrogenesis but the material method section is not very clear to me.
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