Seeding volume in cell culture flasks - (Mar/25/2005 )
What are the typical seeding volumes you have for these T-25, T-75, T-150 flasks?
THanks for any ideas.
justwonder on Mar 25 2005, 12:10 PM said:
T-25: 6ml, T-75: 15ml, T-150: 30-40 ml according to:
McAtier, J.A. and Daview, J., Basic Cell Culture. A Practical Approach., Davies, J.M. (Ed) p. 153 (Oxford University Press Inc, New York, 1998).
THanks for any ideas.
Hi there,
Do you mean, how much volume of media can you have in T25, T75s?
If so, then you are right.
T25 - about 5mls
T75 - about 10 - 15 mls
T-150 - about 20-40 mls.
The stated values are good volume ranges for adherent cells however, if you are asking for seeding volumes for suspension cultures try ...
T25: 4-12 ml
T75: 8-40 ml (can do 50 ml if required)
T150: 15-100 ml
with cells at 1 x 10e5 to 2 x 10e5 cells/ml.
AussieUSA on Mar 29 2005, 10:37 AM said:
T25: 4-12 ml
T75: 8-40 ml (can do 50 ml if required)
T150: 15-100 ml
with cells at 1 x 10e5 to 2 x 10e5 cells/ml.
will it be ok if the seeding volume is upto 40ml in T-75 flask
In my hands, no.
The reason is when you lay the flask on its side during incubation, the amount of media flows too closely to the neck which could lead to contamination.
For suspension cultures, the largest volume I've used is 16 mL.
If you need larger volume, try a spinner flask or roller bottle.
labrat612 on Feb 25 2009, 08:06 AM said:
The reason is when you lay the flask on its side during incubation, the amount of media flows too closely to the neck which could lead to contamination.
For suspension cultures, the largest volume I've used is 16 mL.
If you need larger volume, try a spinner flask or roller bottle.
yes your are correct i used only 15ml but i saw that we can make upto 40ml
i want to try...as it is not possible to keep the flask flat ...could it be possible to stand the flask ?
do the cells still be happy in that conditions with less length?
Are you dealing with suspension cells?
yes i working with suspension cells
swarna on Feb 25 2009, 08:31 AM said:
labrat612 on Feb 25 2009, 08:06 AM said:
The reason is when you lay the flask on its side during incubation, the amount of media flows too closely to the neck which could lead to contamination.
For suspension cultures, the largest volume I've used is 16 mL.
If you need larger volume, try a spinner flask or roller bottle.
yes your are correct i used only 15ml but i saw that we can make upto 40ml
i want to try...as it is not possible to keep the flask flat ...could it be possible to stand the flask ?
do the cells still be happy in that conditions with less length?
I don't think your cell will be happy.
If you stand the flask, the cell will accumulate at the bottom of the flask and reduce the area for them to grow.
The cells at the very bottom will not get nutrient from the medium. Unless you gently shake the flask every hour.
This is why people invented rolling bottle.
Hope this may help.