Suspension cells - (Feb/14/2007 )
Can anyone tell me the purpose of adding F68 in the medium to get the adherent cells adapted to suspension media? All I know is to protect the cells from the shearing stress in rotation movement. anything else?
I have read in the literature that the surfactant F68 can under some conditions can :-
Enhance cell growth.
Enhance protein production.
Prevent damage to cells under Bioreactor stirring conditions.
The mechanism of action accounting for these observations are unknown.
Others in the literature have used F68 and have found it to ahve no effect. I have never used F68 and would try other methods to reduce cell damage. In my experiments, RPM of stirring is the most important variable when looking at cell viabilty.
Enhance cell growth.
Enhance protein production.
Prevent damage to cells under Bioreactor stirring conditions.
The mechanism of action accounting for these observations are unknown.
Others in the literature have used F68 and have found it to ahve no effect. I have never used F68 and would try other methods to reduce cell damage. In my experiments, RPM of stirring is the most important variable when looking at cell viabilty.
We are trying to adapt adherent cells to suspension medium, hence putting them in shakers. What do you mean by saying RPM of stirring? Do you mean to say that setting the rpm to low? These are not spinners but in shaker flasks. Can you please elaborate?
i searched the web, and found that F68 as a surfactant facilitate the release of cell membranes, reduce adsorption of membrane proteins in particular by decreasing surface tension
take a look here
Enhance cell growth.
Enhance protein production.
Prevent damage to cells under Bioreactor stirring conditions.
The mechanism of action accounting for these observations are unknown.
Others in the literature have used F68 and have found it to ahve no effect. I have never used F68 and would try other methods to reduce cell damage. In my experiments, RPM of stirring is the most important variable when looking at cell viabilty.
We are trying to adapt adherent cells to suspension medium, hence putting them in shakers. What do you mean by saying RPM of stirring? Do you mean to say that setting the rpm to low? These are not spinners but in shaker flasks. Can you please elaborate?
Dear Scifi
We routinely grow our suspension cells in Techne Stirrer bottles which are stirred using specialised sitirrer platforms, again made by Techne. These platforms allow either 2 or upto 4 stirrer flasks to be used at any one time. The bottles range from 125ml to 2 Litres and can be stirred at a Revolution Per Minute (RPM) between 0 to 80 RPM. They also have the capacity to stir on an intermittant mode i.e. 5 minutes stirring every hour (55 mins without stirring). We have used this mode to grow Endothelial Cells on Microcarrier beads. The intermittant mode stops aggregation of the cytodex and also reduces the shear stresses on the cells.
I take it that the shakers you are referring to are orbital shakers ? In my experience use the equipment that is SPECIFICALLY for cells and not shakers that are predominately used for Bacteria.
I have no experience in adapting adherence cells to suspension culture. Have you thought about Cytodex beads for example. The adherent cells attach to the beads and then the beads float in supension. The advantage to this method is to grow vast numbers of cells in a small footprint and also to use the beads/cells, no trypsinisation is required.
I hope this answers some of your questions.