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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

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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