I would like to produce synchronized adherent mammalian cell cultures for RNAseq analysis preferably using automated physical means, so the cells would be in most undisturbed and happy. I assume most of the synchronization means are very stressful for the cells and therefore RNAseq would be polluted with the cell stress response genes.
So I wonder, is there a way to gradually synchronize the cells maybe using programmed daily temperature cycle or CO2 cycle or something gentle of this kind.
Hi Max,
If the cells are primary cell culture, you can syn them by confluence. Simply grow them until confluent, and wait another 1 or 2 days. You may want to change media during the culture to avoid nutrient stress. You can get >90% cells in G0/G1 phase. This is not perfect, but may be the mildest way to syn cells.
Good luck!
Xiao Tian
By allowing cells to get confluent, they will be arrested in G0/G1. However, this synchronisation is product of contact inhibition (anchorage related, cadherins) and expansion severely decrease/ceases. Additionally, cells can suffer modifications or start differentiation and if confluence is sustained, your produced cells will be different from the originals. So be careful.
Alternatively, you can cause reversible arrest in different stages of the cell cycle by using drugs like DMSO.
Have a look to the following papers
http://www.ncbi.nlm.nih.gov/pubmed/10023662
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3694177/
hope this helps!
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