Madin-Darby Bovine Kidney (MDBK) Media
One of the most important and well established systems for the production of vaccines is the growth of attenuated viruses in Madin-Darby kidney cells. Current manufacturing processes utilizing these cells rely on the use of serum supplemented medium. However, the use of serum in a manufacturing process introduces a number of concerns such as material costs, increased down-stream purification complexity, and biological contamination risks.
To address these concerns, Sigma has developed a two-medium serum-free cell
culturing system. Medium M 3553 is designed to provide rapid cell growth, and medium M 0682 is designed to provide high titer virus production in sustained high density cell culture.
MDBK cells can be grown to high densities in M 3553 in 2 to 5 days. At that time, the cells can be transitioned into medium M 0682, and subsequently
inoculated with the virus of interest.
It has also been found the mixing M 3553 and M 0682, in various ratios (i.e.
1:1), gives very favorable growth of MDBK cells and production of virus.
| Product # |
Product Name |
Description |
Application |
|
M 3553 |
Madin-Darby Bovine Kidney Growth Medium Serum-Free (MDBK-GM-SF) |
- Liquid;
- Contains inorganic salts, HEPES and sodium bicarbonate buffers, essential and non-essential amino acids, vitamins,
recombinant human insulin and growth factors, fetuin, transferrin, other organic compounds and trace elements.
- Contains 150 ug/ml of animal-derived protein.
- Without L-glutamine; supplement with 16.5 ml of 200 mM L-glutamine per liter of medium prior to use.
- Sterile-filtered
- Endotoxin tested;
- Cell culture tested
|
Media M 3553 and M 0682 comprise a system designed to support the production of viruses in two phases, the growth phase, and the high density
maintenance and virus production phase. This system provides higher viral titers than serum-supplemented media. M 3553 is formulated to support the optimal growth of Madin-Darby Bovine Kidney (MDBK) cells, in which viruses useful as vaccines can be propagated.
This serum free, low protein medium is designed to grow the MDBK cells before transfer into the protein-free medium, M 0682.
The MDBK cell growth, maintenance and virus production system is recommended for use in roller bottle and microcarrier bead based bioreactor systems. |
|
M 0682 |
Madin-Darby Bovine Kidney Maintenance Medium
Protein-free.
(MDBK-MM-PF)
|
- Liquid
- Protein-free medium containing organic salts, HEPES and sodium bicarbonate buffers, essential and non-essential amino acids, vitamins, recombinant insulin, other organic compounds, and trace elements.
- Without L-glutamine; Aseptically add 20 ml of 200 mM L-glutamine solution per liter of medium prior to use.
- Sterile-filtered
- Endotoxin tested;
- Cell culture tested
|
Media M 3553 and M 0682
comprise a system designed to support the production of viruses in two phases, the growth phase, and the high density
maintenance and virus production phase. This system provides higher viral titers than serum-supplemented media.
M 0682 is formulated to maintain high cell densities of MDBK cells over extended periods of time.
This zero protein medium is designed to optimize virus propagation in cells previously grown in M 3553.
The MDBK cell growth, maintenance and virus production system is recommended for use in roller bottle and microcarrier bead based bioreactor systems.
|
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Madin-Darby Canine Kidney (MDCK) Media
The Madin-Darby canine kidney cell line is an important, well established epithelial cell line. These cells have been used extensively as a model system for
studies of renal function, as well as the host system for the growth of attenuated viruses for vaccine production.
Current manufacturing processes utilizing these cells rely on the use of serum supplemented medium. However, the use of serum in a manufacturing process
introduces a number of concerns such as material costs, increased down-stream
purification complexity, and biological contamination risks. Sigma has developed serum-free and protein-free media that promote the growth of MDCK cells at rates and densities comparable to serum supplemented cultures.
These media offer a number of advantages including improved medium definition, reduced cost, improved down-stream processing, and reduced biological safety risks
associated with using traditional serum-supplemented media. In these media
the use of animal derived protein has been dramatically reduced. All hormones and growth factors used are recombinant proteins that were expressed in non-vertebrate systems.
MDCK Protein-Free medium (M 3678) exhibits growth performance in static cultures comparable to DMEM/F-12 (1:1) medium containing 5% fetal bovine serum (FBS). MDCK Serum-Free medium (M 3803) is supplemented with bovine serum albumin and it is used to facilitate the attachment of the MDCK cells to the surfaces of roller bottles and microcarrier beads in non-static cultures.
| Product # |
Product Name |
Description |
Application |
| M 3803 |
Madin-Darby Canine Kidney Medium, Serum-free
(MDCK-SF) |
- Liquid,
- Contains inorganic salts, essential and non-essential amino acids, vitamins, recombinant human insulin and growth factors, other organic compounds and trace elements.
- Contains 100 ug/ml of bovine serum albumin
- Without L-glutamine; supplement with 16.5 ml of 200 mM L-glutamine per liter of medium prior to use.
- Sterile filtered;
- Endotoxin tested;
- Cell culture tested.
|
Complete serum-free medium formulated
to support the growth of Madin-Darby canine kidney (MDCK) cells which support the production of viruses useful as vaccines.
Regulatory
friendly. The use of processed animal proteins, such as bovine serum albumin, in place of serum greatly reduces the risks of biological
contamination.
Completely animal component free custom formulations are available.
|
| M 3678 |
Madin-Darby Canine Kidney Medium, Protein-free
(MDCK-PF)
Performance Data
|
- Liquid,
- Contains inorganic salts, essential and non-essential amino acids, vitamins, recombinant human insulin and growth factors, other organic compounds and trace elements.
- Without L-glutamine; supplement with 16.5 ml of 200 mM L-glutamine per liter of medium prior to use.
- Sterile filtered;
- Endotoxin tested;
- Insect cell culture tested.
|
Complete protein-free medium formulated to support the growth of Madin-Darby canine kidney (MDCK) cells which support the production of viruses useful as vaccines.
Regulatory friendly. Completely free of biological contamination risks associated with the use of animal derived proteins.
Completely animal component free custom formulations are available.
|
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Titerhigh™ Chick Embryo Fibroblast (CEF) Medium
Chick embryo fibroblast cells are used in a number of research applications. In addition, they are the cell line of choice for the production of a number of viruses which are used to manufacture vaccines.
The vaccine market is highly competitive and price sensitive. A major cost factor in the production of vaccines using CEF cells has been fetal bovine serum (FBS). The following formulation has been developed to allow for the use of less expensive iron-supplemented calf serum in lieu of FBS.
| Product # |
Product Name |
Description |
Application |
|
C 3978 |
TiterhighTMChick Embryo Fibroblast
(CEF) Medium |
- Liquid;
- Proprietary formulation.
- Contains inorganic salts, bicarbonate buffer, essential and non-essential amino acids, vitamins, other organic compounds and trace elements.
- Without phenol red, antibiotics, antimycotics, or L-glutamine; supplement with 20 ml of 200 mM L-glutamine per liter of medium prior to use.
- Sterile filtered;
- Endotoxin tested;
- Cell culture tested.
|
Optimized for commercial virus production in chick embryo fibroblasts or fibroblast-like cells.
Comparable growth and virus production results are achieved when iron-supplemented calf serum (0.5-2.0%) is used
in place of FBS. This may result in significant cost savings.
Performs exceptionally well when used in microcarrier bead adherent cell culture systems. Even at serum concentrations as low as
0.5-2%. |
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Vaccine Companion Products and Medium Supplements
This table is a listing of Sigma products that may be used at various times in the processing of cells used for vaccine production.
Details for the use of these products may be found in literature published by Sigma which discusses the use of cell lines such a MDBK, MDCK and CEF for vaccine manufacturing applications.
| Product # |
Product Name |
Description |
Application |
| A 1345 |
Anti-oxidant Supplement (1000x) |
- Liquid
- Sterile filtered.
- Endotoxin tested.
- Cell culture tested.
|
Proprietary medium supplement specially formulated to boost cell growth for enhanced protein expression. May be used in combination
with Polyamine Supplement (P 8483)
|
| C 6295 |
Cell Freezing Medium-DMSO
Serum-free |
- Liquid, frozen
- Sterile filtered.
- Endotoxin tested.
- Cell culture tested.
|
Complete ready-to-use medium containing Minimum Essential Medium, dimethyl sulfoxide (8.7%) and methylcellulose.
Prepared according to the formulation of Waymouth (TCA Manual 2(1), 311-313 (1976). Use to cryo-preserve cells.
|
| D 8537 |
Dulbecco's Phosphate Buffered Saline Modified |
- Liquid
- Without calcium chloride and magnesium chloride
- Cell culture tested
- Sterile-filtered
- Endotoxin tested
|
Calcium and magnesium promote cell adhesion. Use this formulation of D-PBS when tissue
or cell dissociation or release is the objective.
|
| D 8662 |
Dulbecco's Phosphate Buffered Saline |
- Liquid
- With calcium chloride and magnesium chloride
- Cell culture tested
- Sterile-filtered
- Endotoxin tested
|
Known as D-PBS, this is a general use, isotonic saline solution for washing cells
and tissues. |
| G 7513 |
L-Glutamine Solution, 200 mM |
- Liquid
- Sterile filtered.
- Endotoxin tested.
- Cell culture tested.
|
Prepared in cell culture grade water. Useful for supplementing liquid CHO media.
|
| H 9394 |
Hanks' Balanced Salt Solution Modified |
- Liquid
- With sodium bicarbonate
- Without calcium chloride, magnesium sulfate,
- Sterile-filtered
- Endotoxin tested
- Cell culture tested
|
Same as H 9269, except that calcium and magnesium have been removed.
Calcium and magnesium support cell adhesion.
Recommended for washing adherent cells during passage.
|
| P 8483 |
Polyamine Supplement (1000x) |
- Liquid
- Sterile filtered.
- Endotoxin tested.
- Cell culture tested.
|
Medium supplement formulated to boost cell growth for enhancement of protein expression.
|
| T 3924 |
Trypsin-EDTA Solution (1x) |
- Liquid, frozen
- Contains 0.5 grams of porcine trypsin and 0.2 grams of EDTA-4Na per liter of Hank's Balanced
Salt Solution with phenol red.
- Trypsin powder is negative for parvovirus by 9 CFR.
- Sterile filtered.
- Cell culture tested.
|
Use to release adherent cells from supporting surfaces during passage.
|
| T 6414 |
Trypsin Inhibitor Solution (1x) |
- Liquid, frozen
- Contains chromatographically purified soybean trypsin inhibitor and iron supplemented calf serum.
- Prepared in Dulbecco's Phosphate Buffered Saline without calcium or magnesium.
- Sterile filtered.
- Endotoxin tested.
- Cell culture tested.
|
Use to arrest the enzymatic activity of trypsin during passage of adherent cell lines.
|
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