When I read through the method of some journals, they will mention the number of cells they used, for example 1x10^6 cells for immunoblotting.
My question is how do we ensure that there is 1x10^6 cells? As what I (or we) usually do is to plate 0.3x10^6 cells into 6 wells plate and incubating them for overnight. How do we know that in the next day, the number of cells will reach 1x10^6? How do we know that all cell lines are growing at the same rate?
Hello Wai Kit Cheng,
Cell division can be evaluated by the cell doubling time. As an example, if you plate 0.3x10^6 cells you ll get approximately 0.6x10^6 after 24 hours but this is depends on culture conditions and cell health.
Each cell line has its own doubling time.
Hope it helps.
Best,
Hamadi
Hi Wai,
You may not have to worry so much about the number of cells as in any case you will quantify your total protein before loading, and then load the same amount regardless your experimental conditions.
Hi Hamadi, I heard of doubling time before in some of the cell culture textbook, but I still do not understand the meaning of it.. Do you mind explaining to me in layman term?
My understanding is the time for them to double the cell numbers.
Hi Norbert, yes. You got the point. I will run bradford assay to determine the concentration of my protein
Dear Wai Kit Cheng,
Cell doubling time can be calculated in the following way using growth rate (amount of doubling in one unit of time):
- Growth rate:
- N(t)=N(0) e^gr*t
- N(t) = the number of cells at time t
- N(0) = the number of cells at time 0
- gr = growth rate
- t = time (usually in hours)
- For example: the doubling time for T24 cell line(bladder cancer cell line) is approximately 24 hours.
This parameter is very important for experiments because each cell line has it own doubling time.
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