Each Advanced medium is the standard published basal formulation, supplemented with NEAA and sodium pyruvate. We’ve also added these ingredients to allow for serum reduction:
- Ethanolamine
- Glutathione (reduced)
- Ascorbic acid phosphate
- Insulin
- Human (holo) transferrin
- AlbuMAX albumin (a lipid-rich bovine serum albumin)
- Trace salts sodium selenite, ammonium metavanadate, cupric sulfate, and manganese chloride
Actual serum concentration reductions will vary with the cell line and basal medium used.
While the recommended concentration of serum is 1–5%, the concentration must be adjusted for each individual cell line to obtain optimal results. For most applications, no weaning procedures are necessary to reduce serum supplementation by at least 50%. The conversion is simple:
- Centrifuge the cells
- Decant the supernatant
- Resuspend the cells in reduced-serum supplemented medium
For FBS reductions greater than recommended levels, we suggest you titer your FBS concentration to determine maximum reduction. In some cases, additional serum reduction may be obtained by minimal weaning. If you are using antibiotics to control contamination, it is important to reduce the quantity to correspond with the reduced amount of serum. For example, if you reduce serum by half, then reduce antibiotics by half; if you reduce the serum 10-fold, then reduce the antibiotics 10-fold. No reduction is required for selective antibiotics.
Applications for Advanced DMEM/F-12
Cell types | Recommended seed density (viable cells/cm2) | %FBS in Advanced DMEM/F12 |
---|---|---|
Vero | 1.0 x 105 | 0.5–2 |
Jurkat | 5.0 x 108 | 1–2 |
Sp2/01-Ag | 1.5 x 105 | 0.5 |
MRC-5 | 5.0 x 105 | 2 |
WI-38* | 5.0 x 105 | 2 |
*For best results, use Advanced MEM (Cat. No. 12492-013 ) with the WI-38 cell line. |
Applications for Advanced RPMI 1640
Cell types | Recommended seed density (viable cells/cm2) | %FBS in Advanced RPMI |
---|---|---|
Vero | 1.0 x 105 | 0.5–2 |
Jurkat | 5.0 x 108 | 0.5–2 |
Sp2/01-Ag | 1.5 x 105 | 0.5 |
Raji | 1.5 x 106 | 0.5 |
Daudi | 1.0 x 106 | 0.5 |
Applications for Advanced DMEM
Cell types | Recommended seed density (viable cells/cm2) | %FBS in Advanced DMEM |
---|---|---|
A549 | 2.5 x 105 | 1–2 |
COS-7 | 2.0 x 105 | 1–2 |
MDBK | 1.0 x 105 | 1–2 |
MRC-5 | 5.0 x 105 | 2 |
VERO | 1.0 x 105 | 1–2 |
Applications for Advanced MEM
Cell types | Recommended seed density (viable cells/cm2) | %FBS in Advanced DMEM |
---|---|---|
HEp-2 | 3.0 x 105 | 2 |
MDCK | 1.0 x 105 | 2 |
PK-15 | 2.0 x 105 | 1–2 |
VERO | 1.0 x 105 | 1–2 |
WI-38 | 4.0 x 105 | 2 |
For Research Use Only. Not for use in diagnostic procedures.