We are growing these cells in 10% serum in RPMI media. After one week of sub-culturing these cells, they suddenly started clumping .Can anyone suggest how to avoid these clumps?
Tho' m not working on either, I used to get clumps in my cell type (HepG2) too.. We got rid of it over time by trying to make single cell suspensions while seeding.. C if it works for u too..
I also work on HepG2. Try pipetting the cells quite vigorously before plating, perhaps you could even try using a cell strainer. Also try a density titration, I found my cells tend to clump when 1) they are plated at very low densities 2) they are plated from (too) high density cultures 3) they are stressed or are a high passage.
Thanks for response.I have tried vigorous pipetting to get rid of these clumps and to make single cell suspension.But after 12 hours of pipetting again these cells are started to clump.i used to put these cells in 0.3x106 cells per ml as per literature of these cell lines.So any one suggest me what can be the possible factors that is responsible for clumping,so that i will try to avoid it or what else we add in supplementing media which can inhibit clumps.
Thanks :)
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