Learn More
Gibco™ FreeStyle™ 293 Expression Medium
Beschreibung
Gibco FreeStyle 293 Expression Medium is an animal origin–free, chemically defined, protein-free medium specifically developed to support the growth and transfection of 293-F cells in suspension without adaptation. The medium is complete and ready-to-use, with no supplementation required.
Features of FreeStyle 293 Expression Medium include:
- Complete and ready-to-use format containing GlutaMAX supplement
- Ability to support suspension growth and transfection of 293-F cells
- Animal origin–free, protein-free and chemically defined formulation
- Easy use with the FreeStyle 293 expression systems
- Scalability in spinner flasks and bioreactors
Complete and ready-to-use format containing GlutaMAX supplement
FreeStyle 293 Expression Medium is ready to use. It does not require the addition of serum, glutamine, or surfactants. It contains GlutaMAX supplement to minimize toxic ammonia build-up and improve cell viability and growth in an easy-to-use format.
Ability to support suspension growth and transfection of 293-F cells
Gibco 293-F, 293-H, and FreeStyle 293-F cells in suspension can be cultured directly into FreeStyle 293 Expression Medium without adaptation. Cells adapted to other commercially available serum-free media can be subcultured directly into FreeStyle 293 Expression Medium, usually without any further adaptation. Cells usually require adaptation from serum-containing formulations. Cells cultured in FreeStyle 293 Expression Medium can be transfected using lipid based transfection reagents, such as 293fectin reagent (Cat. No. 12347019) and Freestyle MAX reagent (16447500).
Animal origin–free, protein-free and chemically defined formulation
FreeStyle 293 Expression Medium is animal origin–free, protein-free and chemically defined, allowing for easier purification of your protein of interest. Gibco chemically defined media contain no proteins, hydrolysates, or components of unknown composition.
Easy use as part of the FreeStyle 293 Expression System and FreeStyle MAX Expression System for protein production
The FreeStyle 293 Expression System (Cat. No. K900001) is a complete, suspension cell culture system for generating large amounts of mammalian recombinant protein (see figure). It combines FreeStyle 293 Expression Medium with 293fectin reagent, a cationic-lipid formulation designed to transfect suspension FreeStyle 293-F cells at high efficiency. The FreeStyle MAX Expression System is a breakthrough technology for rapid and high-yield mammalian protein production. It combines FreeStyle media, FreeStyle MAX Reagent, and either FreeStyle CHO-S cells or FreeStyle 293-F cells.
Scalability in spinner flasks and bioreactors
Protein production using FreeStyle 293 Expression Medium can be scaled up in spinner flasks or bioreactors. The appropriate spinner or impeller speed and seeding density should be optimized for each system. Depending on the impeller design and speed, it may be necessary to supplement the medium with additional Pluronic F-68 to avoid sheer stress in the culture.
Universal bag packaging
Gibco universal bags offer flexible packaging that allows you to integrate a single-use, disposable large volume media container with a wide variety of automated cell culture instruments, disposable bioreactor systems, and sterile connection processes, minimizing the need for costly customization.
Product use
Customers using FreeStyle 293 Expression Medium in a manufacturing process, who have a submission with the FDA, may request a letter of authorization from us to reference our Type II Drug Master File (DMF).
cGMP manufacturing and quality system
FreeStyle 293 Expression Medium is manufactured at a cGMP-compliant facility located in Grand Island, New York. The facility is registered with the FDA as a medical device manufacturer and is certified to ISO 13485 standards.
Spezifikation
Spezifikation
| Cell Line | FreeStyle™ 293-F Cells |
| Classification | Animal Origin-free, Chemically-defined, Protein-free, Serum-free |
| Form | Liquid |
| Product Type | Expression Medium |
| Serum Level | Serum-free |
| Product Line | Freestyle |
| Quantity | 1 L |
| Shipping Condition | Room Temperature |
| Content And Storage | Storage conditions: 2°C to 8°C. Protect from light Shipping conditions: Ambient Shelf life: 12 months from date of manufacture |
| Culture Type | Suspension Cell Culture |
Frequently Asked Questions (FAQs)
With FreeStyle 293 Expression Medium, we would recommend harvesting the cells a little earlier to reduce any nonspecific binding by host cell proteins and then filtering the culture supernatant through a 0.2 or 0.1 µM membrane before loading onto the column. On the other hand, FreeStyle CHO Expression Medium contains EDTA that will strip the nickel column. The medium can be subjected to dialysis or buffer exchange prior to loading on the column, or the EDTA can be chelated by adding 1-3 mg/L NiSO4 or NiCl to the supernatant. Iron or cobalt salts can also be used.
You should be able to use the ExpiFectamine293 Reagent for transfection of Freestyle 293-F cells grown in FreeStyle 293 Expression Medium; however, the enhancers will provide only a little boost in expression as they are designed to work with higher density cultures that FreeStyle 293 Expression Medium cannot support.
When cells meant to be grown in suspension are grown in static culture, they may form clumps. These clumps will severely limit transfection efficiency and protein expression. It is suggested that FreeStyle 293 cells in FreeStyle media and CHO-S cells in CD-CHO or CHO-SFM are grown in agitated suspension to reduce the appearance of clumps. However, if clumps do form, you can try the following protocol to select for cells that don't form clumps:
- Transfer cells into an appropriate size centrifuge tube that will hold the entire cell suspension.
- Allow cells to sit undisturbed for about 5 minutes. The time can vary depending on the specific cell line. Larger cell clumps will settle to the bottom of the tube.
- Collect cells from the upper portion of the tube to passage into a new flask.
Our recommendation of 8% CO2 for the optimal growth of Freestyle 293 cells in Freestyle 293 Expression Medium is based on the bicarbonate content in the media and growth conditions of the cells observed by R&D. That said, you may use lower concentrations of CO2 if necessary (5% is common) to grow these cells. You may observe slightly slower growth kinetics or lower cell concentrations, but functionally the entire system will still work.
Yes, the FreeStyle 293 Expression Medium contains Pluronic F-68 and lipids that would interfere with ammonium sulfate precipitation. Dialysis or tangential flow filtration can be used to remove lipids and Pluronic F-68 from the medium. Then you can concentrate the medium with ammonium sulfate.
For Research Use or Further Manufacturing. Not for diagnostic use or direct administration into humans or animals.
Indem Sie auf Absenden klicken, erklären Sie sich damit einverstanden, dass Fisher Scientific sich mit Ihnen in Verbindung setzen kann, um Ihr Feedback in diesem Formular zu bearbeiten. Wir werden Ihre Informationen nicht für andere Zwecke weitergeben. Alle bereitgestellten Kontaktinformationen werden in Übereinstimmung mit unserer Datenschutzrichtlinie aufbewahrt. Datenschutzrichtlinie.