is the media requirement and seeding cell density same for the suspension as for the adherent cells.
I am working with T cell and I am doubtful about how of media to be added and what is seeding cell density.
Suspension and adherent cell line are usually not exactly the same for seeding density.
It depends on the cell line and how fast they proliferate. 1x10e5 cells/mL is probably ok.
What kind of the cells (which cell line) and what media are you using? It is important to consider how fast they double it's number. Recommend to use a media with pH indicator, helps to know the best density. For example; jurkat cells in RPMI can be cultured at low density, 0.200x10^6 in 10mL dish.
Ok! That cells are harder to grow. It is better to seed them between 1-2.5x 10^5 cells/mL . Keep that concentrations also post activation. I assume you are using FBS at 10%, but if it is possible and if the cells are human, try to get a serum free media, but it is not absolutely necessary.
Xochitl Ambriz Peña
by the term 1-2.5x 10^6 cells/ml, do you mean to say that in suspension culture one has to look for the volume of media rather than looking for the surface area of culture vessel used. hence it is that I should keep 1-2.5x 10^6 cells/ml this standard value and use it according to the volume of media added.
And yes, I am also aware of the fact that volume of media is dependent on the volume of culture vessel.
I actually want to know how should one decide how much cells to be inoculated.
my cells are growing well with CD3/CD28 mAB, but the cell density was less.
thank you
Harish, at any final volumen you are working you can adjust the number of cells, just keep the density
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