Q. How does FUJIFILM Irvine Scientific define serum-free, animal-component free, protein-free, and chemically-defined media?
A. Here is how FUJIFILM Irvine Scientific defines these types of media:
- Serum-free media do not contain or require the addition of serum.
- Protein-free media do not contain any proteins but can contain undefined peptides from plant hydrolysates.
- FUJIFILM Irvine Scientific animal-component free products are manufactured with raw materials that do not contain any animal- or human-derived products to the tertiary level and are manufactured using equipment that does not come in contact with any animal or human products. The raw material handling, storage and final product manufacture and packaging are performed in dedicated human- and animal-component free facilities and equipment.
- FUJIFILM Irvine Scientific chemically-defined media do not contain any undefined animal-derived raw materials and are free of any animal-derived components, including human plasma or human blood-derived components. All raw materials are defined and their exact concentrations are known, which can include recombinant proteins.
Q. Why should I use serum-free/chemically-defined media?
A. Serum-free and chemically-defined media allow for a more consistent culture system (reducing lot-to-lot variation) because they do not contain undefined components that may be introduced by the addition of serum. Serum-free media are lower in protein content than medium supplemented with serum, which can simplify the purification process and increase the yield of the end-product. In addition, the end-user also does not need to take time to qualify and secure lots of FBS for activity.
Q. What does FUJIFILM Irvine Scientific have to offer in terms of serum-free, animal-component free, and chemically-defined media?
A. FUJIFILM Irvine Scientific offers many serum-free media, including products for three main types of cells currently used for bioproduction and bioprocessing: Chinese Hamster Ovary (CHO) cells, HEK293 cells, and Hybridoma cells. Within each cell line, we offer a suite of growth media and feeds.
BalanCD CHO Growth Medium and Feeds are chemically-defined, animal-component free and formulated without L-glutamine, Hypoxanthine, and Thymidine to be used in a variety of applications. BalanCD CHO Feeds are specifically designed to increase process yields of antibodies and recombinant proteins in CHO cells.
- BalanCD CHO Growth A Medium - Liquid
- BalanCD CHO Growth A Medium - Powder
- BalanCD CHO Feed 1 - Liquid
- BalanCD CHO Feed 1 - Powder
- BalanCD CHO Feed 2 - Liquid
- BalanCD CHO Feed 2 - Powder
- BalanCD CHO Feed 3 - Liquid
- BalanCD CHO Feed 3 - Powder
- BalanCD CHO Feed 4 - Powder
- Anti-Clumping Supplement
BalanCD Transfectory CHO System is chemically-defined and animal-component free. The system is designed for transient transfection in CHO cells to achieve higher titers and ensure process consistency.
- BalanCD Transfectory CHO - Liquid
- Transfectory Supplement, contains animal-component free concentrated hydrolysate supplement
- Anti-Clumping Supplement
The BalanCD HEK293 system platform includes a growth medium, feed and anti-clumping supplement. The scalable platform, which is chemically-defined and animal-component free, is optimized for the production of viral vectors and transient protein expression.
IS MAB-CD is a chemically-defined, low protein medium for the growth of hybridoma and myeloma cell lines for recombinant monoclonal antibody production. This chemically-defined medium is regulatory friendly and allows for improved lot-to-lot consistency compared to other formulations that are not chemically-defined.
Q. When adapting cells from serum-containing to serum-free media, is the direct or sequential adaptation method better?
A. The sequential adaptation method is recommended. Sequential adaptation allows the cells to gradually adapt to their new serum-free environment. Below is an example protocol. Cell types vary, and contact us for a tailored solution.
Suggested Sequential Adaptation Protocol:
- Start with cultures (suspension or adherent) that are at maximum cell density and high viability (>80%). If cells are attachment dependent, trypsinization procedures should be used.
- Split cells 1:2 using the desired final serum-free medium as the diluent.
- Incubate cells until the maximum cell density is achieved.
- Split cells again. The ratio may vary depending on whether the cells are growing attached or in suspension culture. For attached cells, try 1:2 or 1:3 split based on confluency. For suspensions, try 1:3 to 1:5 or approximately 3-4x105 cells per mL.
- Continue incubation until optimal cell density is achieved. (Maximum may imply over confluency which should be avoided generally.)
- Monitor the growth and viability of the cells relative to cells cultured in the original or control medium. Cells that have been successfully adapted to serum-free media should have the viability greater than 85%, with a doubling time equivalent to the control medium (serum-containing or other serum-free medium) from which the culture was adapted.
- Low viability or poor doubling times indicate that the culture may not yet be adapted. 3 to 5 successive splits using 1:2 to 1:3 may be necessary. It is not uncommon for some cells to continue to require 0.1 to 0.5% serum.
- Time for adaptation to serum-free conditions will vary with the growth characteristics of the particular cell type. If at any time during the weaning process the culture viability drops below 80%, or the time to double increases significantly, weaning may have occurred too quickly. The cells may require several passages at the previous dilution before renewal of the sequential weaning process resumes.
Q. Why was L-Glutamine eliminated from many of your formulas?
A. L-Glutamine is easily hydrolyzed in liquid and byproducts of hydrolysis can be toxic to the cell. Additionally, the necessary concentration of L-Glutamine may be compromised after a period of time. Since L-Glutamine is an essential amino acid, it is best for the end users to supplement media with the recommended concentration of fresh L-Glutamine.
Q. How do we prevent mycoplasma contamination in cell culture?
A. Good aseptic techniques should be used in all laboratory applications. The best method for prevention is to test all materials that come in contact with the cells. This should be done on a regular basis to ensure that mycoplasma is not introduced into the cell culture and to identify any potential sources of contamination.
Q. What are the minimum requirements for a custom media formula?
A. We generally require a 100L minimum order to manufacture a custom formula. Our Manufacturing Support and Service (MSS) can make smaller non-GMP quantities to meet special investigational or development needs for our customers.
Q. What is the timeline on a custom formula?
A. Customs are usually delivered to the customer between 8 to 10 weeks after we receive the final signed documents with a purchase order.