Serum-Free Media for Animal Cell Culture in Suspension

The use of serum-free media brings many advantages including: batch to batch uniformity, consistency in prices, freedom from concern regarding mycoplasma and other contaminants, as well as simplified product purification and downstream processing.

BI's serum-free media and supplement have been proven to be effective for a wide variety of applications such as:
  • Culture of murine myeloma cells (NS-1, NS-0/1, SP2)
  • Culture of mouse-mouse hybridoma cells (from NS-1, NS-0/1, SP2, X63-Aag8-653, BW-S147)
  • Culture of rat-mouse, rat-rat, and human-human hybridomas
  • Monoclonal antibody production
  • Culture of other human, murine and avian lymphocytes
  • Viral production (such as HEV virus from Turkey B-lymphoblastiod cell lines)
  • Culture of various anchorage-dependent cells (such as human osteosarcoma, human epithelial, and thyroid cells)
  • Maintenance of most anchorage-dependent cells during long production periods
DCCM-1: A totally defined high protein serum-free medium, designed for hybridoma cell growth and monoclonal antibody production. The product is ready to use and needs only the addition of L-Glutamine.

DCCM-2: The same as DCCM-1 but with low protein formulation. Glutamine must be added prior to use.

LPM (BSA-free): This formulation is free of Bovine Serum Albumin. Despite this very low protein content, LPM has proven to be remarkably effective for the growth of a wide variety of hybridomas and other lymphocytes.
Glutamine must be added prior to use.
Growth of Hybridoma 3D2 C2 (mouse-mouse from Myeloma X63-Ag 8-653)

hybridoma growth in serum-free media

Growth of Hybridoma 4-15 (mouse-mouse from Meyloma NS-1Ag 4.1)

growth of hybridoma in serum-free media

BIOGRO-2 Serum-free media supplement product
In addition to our line of ready to use serum-free media products, BIOGRO-2 (05-610-1) for those customers who prefer to prepare final media on their own.
The BIOGRO-2 formulation is totally free of serum albumin, and the concentration of total protein will therefore be less than 25 micrograms per ml in the final medium.
Method of use:
BIOGRO-2 is 50 fold concentrates and therefore are recommended for use at a concentration of 2% (although in some cases 1% may be sufficient). DMEM F12 (1:1) has found to be most generally effective as the basal media, but many cell lines grow well with RPMI medium, DMEM or IMDM. Glutamine and antibiotics should be added to the final formulation.

Cell Adaptation to serum-free medium

For many cell types no adaptation procedures are necessary and may even be detrimental. In other cases standard-weaning procedures may be necessary.

Sequential adaption
  • Subculture the cells growing in serum-supplemented medium into 1:1 (v/v) mixture of serum-free medim and serum supplemented medium.
  • When the cell density is >5x105 cells/ml, subculture the cells in an equal volume of SFM at a cell density 2.5x105 to 3x105 cells/ml.
  • Continue to subculture after the cell density is >5x105 cells/ml in serum-free medium until the serum is 0.1% with about 85% cell viability.
  • Subculture the cells into SFM with an inoculum of 2.5x105 to 3x105 cells/ml.
  • When the cell density is 1x106 to 3x106 cells/ml (4 to 6 days post planting) subculture the cells again.
  • Stock cultures of serum-free medium adapted cells should be subcultured is serum-free medium every 3 to 5 days when the cell density is 1x106 cells/ml with 90% viability.
Direct adaptation
Some cells can be directly adapted from serum containing medium to serum-free medium. For direct adaptation, the cell inoculum should be 1.5x105 to 3x105 cells/ml. Cells should be subcultured when the cells density is 1x106 cells/ml. Cells are fully adapted to SFM when the cell density is 2x106 to 4x106 cells/ml after 4 to 7 days in culture. Stock cultures of cells adapted to serum-free medium should be subcultured in serum-free medium every 3 to 5 days when the cell density is 1x106 to 3x106 cells/ml with 90% viability.

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