FIBERCELL™ FAST FACTS

The primary applications for hollow fiber cell culture are:

•  Monoclonal antibody production

•  Recombinant protein production

•  Adenoviral vector production

Other applications include:

•  Cytokine and growth factor production

•  Lymphocyte expansion

•  Endothelial cell culture under shear stress.

Generally, FiberCell™ hollow fiber cell culture modules are used to MAKE something of biological interest. Cells can be grown using standard mediums, however low glucose mediums such as RPMI should be avoided as there is not sufficient glucose to support high density culture. Any medium adaptations can be performed in the cartridge.

Monoclonal antibody production

Cat # 4300-C2011 or 4300-C2018 should be used. This MWCO allows TGF Beta to diffuse away while retaining the produced antibody.

4300-C2011 will support up to 2 X10 9 cells; this is equal to a two-liter culture, 20 roller bottles or 50-60 T175 flasks

° Produce 5-50mg of antibody every 2 days, average is 20mgs per harvest

° Continue to produce antibody for up to 6 months of continuous culture

° Consumes about 1 liter of cell culture medium every two days. To reduce medium consumption harvest more cells out.

° Inoculate 10 8 cells to start.

° A single mouse used for ascites fluid production will produce 10-20mgs total antibody; each harvest is equal to a single mouse.

° Endotoxin burden is 1/10 th that of ascites fluid production.

° Cannot be re-used but can be stored and re-inoculated with the same cell line.

° Hybridomas grown in the FiberCell™ cartridge can be more easily adapted to serum free cell culture medium or adapted to as low as 2% FBS.

 

4300-C2018 Will support up to 5 X10 10 cells; this is equal to a ten to fifty liter culture.

° Produce 50-150mgs of antibody every 2 days, average is 70mgs per harvest

° Consumes about 2 liters of cell culture medium every day.

Recombinant Protein Production

The molecular weight of the protein to be produced determines the fiber MWCO to use. For proteins larger than 100kd use 4300-C2011 or 4300-C2018

For proteins of 20kd to 100kd use 4300-C2008 or 4300-C2003

For adherent cell types such as CHO and 293 cells flush the cartridge with two changes of PBS for at least two days prior to pre-culturing with cell culture medium. Inoculate the cells with 20% FBS in the cell inoculum only.

 

4300-C2011 Has 2200cm 2 of surface area, equal to 12.5 T175 flasks. Because of the way cells grow when attached to a fiber the total number of cells will be equal to 50-60 T175 flasks or 20 roller bottles.

° Cells should be inoculated at a number equal to 50% of the surface area of the fiber.

° If the protein of interest can be trapped by the molecular weight cut-off of the fiber it will reach a concentration 100 times that of the same cell line grown in flask culture when collected from the ECS (extra-capillary space)

° Average productivity is around 100µg/ml (of ECS volume) or 1mg per day or more.

° Proteins greater than 100kd in molecular weight will be trapped by this fiber.

° If the protein of interest is too small to be trapped (like cytokines and cell growth factors) it will reach a concentration of 10 times that of the same cell line grown in flask culture when collected from the reservoir bottle.

° Harvest every day instead of every two days

° Will consume 1 liter of medium every two days.

° Reduction in serum concentration or easier adaptation to serum-free medium makes purification of proteins or identification of low concentration growth factors easier.

 

4300-C2019 Has 1.2m 2 of surface area. This is equal to 68 T175 flasks. Because of the way that cells grow on hollow fibers this will be equal to over 400 T175 flasks or 100-150 roller bottles.

° Medium consumption will be as high as 4 liters per day

° Can harvest 5-10 mg of protein per day

Adenovirus production

Use 2011 cartridge. 293 cells must be adapted to suspension culture. This is done using serum-free medium or specialty suspension 293 cell lines. Infection is performed directly in the cartridge.

° 4300-C2011 will support up to 10 9 cells

° C2011 produces 5-9 X10 11 pfu (plaque forming units i.e. active virus), 10 13 virus particles. about 70 T175 flasks, or 20 roller bottles

° Closed, bio-safe system.

° Cartridges can be re-used for the same virus up to 5 times.

° Please see references

Endothelial Cell Culture 

Use C2025. This cartridge allows extra-cellular matrix proteins to be attached to the fiber. One of the few ways to grow endothelial cells under conditions of chronic shear stress. When grown under these conditions they behave in a much more in vivo like manner. They lay down flat and form a monolayer, form tight junctions, and certain genes are turned on in response to this shear stress that are not expressed in static culture. Please check the reference list on the FiberCell™ Video CD manual for more information. Other cell types such as vascular smooth muscle and brain glial cells can be co-cultivated with these endothelial cells.

° Inner surface area of the cartridge is 75cm 2

° Load with 2 T75 flasks of cells. Excess cells will be collected in the reservoir bottle.

° C4300-2025 will yield about 100µg of total RNA for gene expression analysis.

° Shear stress of 1-20 dynes/cm 2 can be generated.