Cell counting - (Nov/14/2018 )

bob1 on Fri Nov 30 20:11:59 2018 said:

Primary neurons will grow quite slowly I would assume, probably slower than one replication in 24 h - if you know the doubling time, you could do the calculations yourself. I was assuming cultured cell lines rather than primary cells, so the replication figures I gave above don't really apply. Yes, the plating time makes a difference - the longer you give them, the more they will divide. However, I know primary neurons are slow growing and susceptible to damage fairly easily so you want to give them some time to recover after plating before you start drug treatment.

 

Glial cells replicate quite fast, so it could be that you have some glia in there which are replicating more rapidly. 

Thank you. It might be 10 days was too much for them, I could have started earlier. 

This was a mixed culture, so there must be Glial cells in them. Do you know how fast the Glial cells replicate?

A quick search of the literature tells me about 20-24 hours per replication cycle for glial cells from neonates, same time for astrocytes too. It looks like neural progenitor cells replicate faster than that 13-17h/cycle according to one paper I found.

jojo320 on Tue Feb 19 17:56:53 2019 said:

 

Mad Researcher on Thu Nov 15 01:26:00 2018 said:

Hi,
 
I plated my cells at a density of 50,000 cells/well in 1 ml of media.
 
I have treated my cells with drugs and have counted the cells. I wanted to know the cell density after drug treatment, how can I calculate that?
 
Thanks

What would be the unit and should I consider the dilution factor of 1:1 as I used trypan blue

 

The unit would be cells/unit volume. Usually this is considered to be cells per ml. Unless you are talking about cells per area (in a well or flask), in which case cells/cm² or cells/mm² are fine.

Yes, you should include a dilution factor of 2 in the calculation to account for your dilution of the cells with trypan blue