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Expi293F™ Cells (Gibco™)

Expi293F™ human cells are derived from the 293 cell line, and are a core component of the Expi293™ Expression System. They are maintained in suspension culture and will grow to high density in Expi293™ Expression Medium. Expi293F™ cells are highly transfectable and generate superior protein yields compared to standard 293 cell lines in transient protein expression.

Expi293F Cells attributes include:
Suspension culture cells pre-adapted to Expi293™ Expression Medium
Demonstrated higher protein expression levels than other 293 cells adapted to Expi293™ Expression Medium
Exhibit more rapid cell growth and higher culture viabilities than standard suspension-adapted 293 cells
Consistent transient expression performance of Expi293F™ cells for over 40 passages beyond thaw
• Also available from a cGMP bank (Cat. No. 100044202)

Frozen cells are supplied in a vial containing 1 ml of cells at 1 x 107 viable cells/ml in Expi293™ Expression Medium and 10% DMSO. The cells should be thawed directly into Expi293™ Expression Medium. The six-vial multipack eliminates the need to expand and rebank the cells in the lab.

Cells are typically grown in flasks on a shaker platform in a humidified, CO2 cell culture incubator. However, cultures are scalable from volumes of <1 mL in multiwell plates to >10 L in bioreactors. Protein harvest can be performed at 2 to 7 days post-transfection, depending on the protein expressed. For expression of recombinant antibodies, harvest is typically at 5 to 7 days post-transfection. There is no need for culture medium supplementation or batch feeding during expression.

Mock SLC Transporter Cells (Gibco™)

These cells are intended for use as a negative control in experiments utilizing TRANSiPORT solute carrier (SLC) transporter-overexpressing HEK293 cells. These mock cells do not overexpress the SLC transport protein and so can be used to determine levels of endogenous transport activity as well as non-specific binding of compounds being tested.

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GM1006 Human OATP1B3 Transporter Cells
GM1008 Human OCT1 Transporter Cells
GM1013 Human NTCP Transporter Cells
GM1014 Human MATE1 Transporter Cells
GM1015 Human MATE2K Transporter Cells

293 F Cells, in CD 293 (Gibco™)

Gibco® 293-F cells were cloned from the original 293 cell line and adapted to Gibco® CD 293 Medium. The 293 cell line is a permanent line established from primary embryonic human kidney and transformed with sheared human adenovirus type 5 DNA. The E1A adenovirus gene is expressed in these cells and participates in transactivation of some viral promoters, allowing these cells to produce very high levels of protein. Gibco® 293-F cells in Gibco® CD 293 Medium feature:
• Quick thaw into suspension culture directly in Gibco® CD 293 Medium or Gibco® 293 SFM II
• Faster growth, superior transfection efficiencies and high protein expression levels
• Documented lineage from a low passage Master Cell Bank
• Quality and performance testing

Quick thaw into suspension culture directly in Gibco® CD 293 Medium or Gibco® 293 SFM II
Each vial containing 7.5 × 106 cells can be thawed directly into suspension culture in either Gibco® CD 293 Medium or Gibco® 293 SFM II supplemented with 4 mM L-glutamine or GlutaMAX™ reagent. Protocols are provided in the product manual.

Faster growth, superior transfection efficiencies and high protein expression levels
Gibco® 293-F cells were prepared from a clone selected for fast growth in serum-free medium, superior transfection efficiencies and a high level of protein expression. Optimal transfection efficiencies are obtained with Lipofectamine™ 2000 or 293fectin™ transfection reagents.

Documented lineage from a low passage Master Cell Bank
The parental 293 cells were cloned by limiting dilution; the clone selected exhibited fast growth and high transfection efficiency. This clone was then re-cloned by limiting dilution; of these clones, the 293-F clone was then selected for its fast growth in serum free medium. The serum-free Master Cell Banks were prepared at approximately passage 25.

Quality and performance testing
Each lot of Gibco® 293-F cells is tested for cell growth and viability post-recovery from cryopreservation. In addition, the Master Seed Bank has been tested for contamination of bacteria, yeast, mycoplasma and virus and has been characterized by isozyme and karyotype analysis.

Product Use
For Research Use Only. Not for any animal or human therapeutic or diagnostic use. Caution: Handle as potentially biohazardous material under at least Biosafety Level 2 containment. This product contains Dimethyl Sulfoxide (DMSO), a hazardous material. Review the Material Safety Data Sheet before handling.

Viral Production Cells (Gibco™)

Gibco Viral Production Cells are a clonal derivative of the HEK 293F cell line, and are a core component of the LV-MAX Lentiviral Production System. Viral Production Cells have been adapted to a special chemically-defined, serum-free and protein-free LV-MAX Production Medium. They are maintained in suspension culture and will grow to high density in LV-MAX Production Medium. Viral Production Cells are highly transfectable by LV-MAX transfection reagent and generate superior lentiviral production compared to polyethyleneimine (PEI)-based lentiviral production methods.

• Suspension culture cells facilitate high-throughput screening or scale-up
• Higher viral production levels than other suspension HEK 293 cells
• Rapid cell growth and high culture viabilities

Frozen cells are supplied in a vial containing 1 mL of cells at 1 x 107 viable cells/mL in LV-MAX Production Medium and 10% DMSO. The cells should be thawed directly into LV-MAX Production Medium.

The cells are typically grown in flasks on a shaker platform in a humidified, CO2 cell culture incubator. However, cultures are scalable in volume from 96-deep well blocks to 2-L flasks and bioreactors. Lentiviral vector harvest is typically performed at two days post-transfection.

Flp-In™-Jurkat Cell Line (Invitrogen™)

Flp-In™ Cell Lines are designed for rapid generation of stable cell lines that express a protein of interest from a Flp-In™ expression vector. These cells contain a single stably integrated FRT site at a transcriptionally active genomic locus. Targeted integration of a Flp-In™ expression vector ensures high-level expression of your gene of interest. There are six Flp-In™ Cell Lines available for the generation of isogenic stable cell lines and a Flp-In™ T-REx™ Cell Line for the generation of tetracycline-regulated stable cell lines. Flp-In™-CV-1, Flp-In™-293, Flp-In™-BHK, Flp-In™-Jurkat, and Flp-In™-3T3 Cell Lines were created by transfecting the parent cell lines with pFRT/lacZeo and selecting for stable Zeocin™- resistant clones. The Flp-In™-CHO Cell Line was created by transfecting CHO cells with pFRT/lacZeo2 and selecting for Zeocin™-resistant clones. The Flp-In™ T-REx™-293 Cell Line contains pFRT/lacZeo and pcDNA™6/TR (from the T-REx™ System) stably integrated. Co-transfection of the Flp-In™ Cell Lines with a Flp-In™ expression vector and the Flp recombinase vector, pOG44, results in targeted integration of the expression vector to the same locus in every cell, ensuring homogeneous levels of gene expression.

Choosing your Flp-In™ Vector/Cell Line Combination
The Flp-In™-CV-1, Flp-In™-293, Flp-In™-CHO, and Flp-In™-Jurkat Cell Lines (Figure 1) work well with Flp-In™ vectors that express a gene from the CMV promoter (pcDNA™5/FRT, pcDNA5/FRT/V5-His-TOPO®, and pSecTag/FRT/V5-His-TOPO®). Flp-In™-BHK and Flp-In™-3T3 cells tend to down regulate the CMV promoter. Therefore, it is recommended that the Flp-In™ vectors containing the EF-1α promoter (pEF5/FRT/V5-DEST™ and pEF5/FRT/V5-D-TOPO®) be used with these cell lines.

293A Cell Line (Invitrogen™)

The 293A Cell Line is a subclone of the 293 Cell Line and has a relatively flat morphology. It facilitates the initial production, amplification and titering of replication-incompetent adenovirus. The cell line contains a stably integrated copy of the E1 gene that supplies the E1 proteins (E1a and E1b) required to generate recombinant adenovirus. The flat morphology of the cells makes the titering procedure easier.

Advantages
• Has flat cell morphology suitable for screening adenoviral constructs

Key Features
• Contains a stably integrated copy of the E1 gene that supplies the E1 proteins (E1a and E1b) required to generate recombinant adenovirus.

Kit includes
• 293A Cell Line provided as a frozen stock in 1 ml of 90% complete media and 10% DMSO

FreeStyle™ CHO-S Cells (Gibco™)

FreeStyle™ CHO-S Cells are part of the FreeStyle™ MAX Expression System, which is a breakthrough technology for rapid and high-yield mammalian protein production. This cell line is adapted to high density, serum-freesuspension culture in FreeStyle™ CHO Expression Medium and iscapable of producing high levels of recombinant protein.

ExpiCHO-S™ Cells, 6-vial "Cell Bank" pack (Gibco™)

ExpiCHO-S™ Cells are derived from a non-engineered subclone that has been screened and isolated from CHO-S Chinese hamster ovary (CHO) cells (1). They are a core component of the ExpiCHO™ Expression System Kit (Cat. No. A29133). ExpiCHO-S Cells are maintained in suspension culture, have minimal tendency to clump, and grow to high density in ExpiCHO™ Expression Medium (Cat. No. A2910001). As part of the ExpiCHO Expression System Kit, the cells generate superior protein yields by transient transfection compared to standard CHO, 293, and high-density Expi293F™ cells.

ExpiCHO-S™ Cells:
• Allow you to attain yields of up to 3 g of protein per L of transfected culture as part of the ExpiCHO expression system
• Enable rapid, high-yield transient protein production, allowing labs developing protein biotherapeutics to work with CHO-expressed proteins from start to finish in the drug development process
• Are adapted to high-density suspension growth in ExpiCHO Expression Medium
• Exhibit rapid doubling times of approximately 17 hours
• Enable consistent transient expression performance for over 30 passages after thaw
• Maintain high cell viability and productivity for greater than 10 days post-transfection as part of the ExpiCHO expression system

Frozen cells are supplied in a vial containing 1 mL of cells at 1 x 107 viable cells per mL in ExpiCHO Expression Medium and 10% DMSO. The cells should be thawed directly into ExpiCHO Expression Medium. The 6-vial multipack eliminates the need to expand and bank the cells in the lab.

Cells are typically grown in flasks on a shaker platform in a humidified, CO2 cell culture incubator. However, cultures are scalable from volumes of <1 mL in multi-well plates to >10 L in bioreactors. Protein harvest can be performed at 5 to 14 days post-transfection, depending on the protein expressed.

References
1. Wurm, FM. CHO Quasispecies—Implications for Manufacturing Processes (2013) Processes 1:296-311.

Human OATP1B1 SLC Transporter Cells (Gibco™)

TRANSiPORT Human OATP1B1 SLC Transporter Cells are HEK293 cells that transiently overexpress the OATP1B1 solute carrier (SLC) transport protein. Solute carriers are a super-family of membrane transporters that can affect pharmacokinetics and drug exposure by governing the transport of solutes in and out of cells. The OATP1B1 SLC transporter protein is found in hepatocytes.

• Assess potential for transporter-mediated drug metabolism
• Easy to use format
• Obtain high quality results with a large signal-to-noise ratio

Choice of measurement system
Cell-based assays can be performed using radioisotope-labeled compounds, fluorescence-labeled compounds, or non-labeled compounds. The amount of substrate transported into the cells can be measured directly using a liquid scintillation counter, fluorescence plate reader, or LC-MS/MS, thereby allowing direct evaluation of SLC transporter activity.

Assay reliability
Mock HEK293 cells (Cat. No. GM1001) that do not overexpress the SLC transporter are available for use as a negative experimental control. Some compounds may demonstrate high background levels of transport due to the presence of endogenous transporters or non-specific binding.

Rapid results
The convenient product format enables data generation in 2 days from thawing of the cells to final results.

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GM1003 Human OAT1 Transporter Cells
GM1004 Human OAT3 Transporter Cells
GM1005 Human OCT2 Transporter Cells
GM1006 Human OATP1B3 Transporter Cells
GM1008 Human OCT1 Transporter Cells
GM1013 Human NTCP Transporter Cells
GM1014 Human MATE1 Transporter Cells
GM1015 Human MATE2K Transporter Cells
GM1001 Mock Cells

T-REx™-293 Cell Line (Invitrogen™)

T-REx™ Cell Lines stably express the tetracycline repressor protein (Table 1). They save significant time and effort when using the T-REx™ System. The T-REx™ Cell Lines are functionally tested by transient transfection with the positive control vector pcDNA™4⁄TO⁄lacZ. T-REx™ Cell Lines exhibit extremely low basal expression levels in the repressed state and high expression upon induction with tetracycline (Figure 1).

Human OAT1 SLC Transporter Cells (Gibco™)

TRANSiPORT Human OAT1 SLC Transporter Cells are HEK293 cells that transiently overexpress the OAT1 solute carrier (SLC) transport protein. Solute carriers are a super-family of membrane transporters that can affect pharmacokinetics and drug exposure by governing the transport of solutes in and out of cells. The OAT1 SLC transporter protein is found in kidney proximal tubule cells.

• Assess potential for transporter-mediated drug metabolism
• Easy to use format
• Obtain high quality results with a large signal-to-noise ratio

Choice of measurement system
Cell-based assays can be performed using radioisotope-labeled compounds, fluorescence-labeled compounds, or non-labeled compounds. The amount of substrate transported into the cells can be measured directly using a liquid scintillation counter, fluorescence plate reader, or LC-MS/MS, thereby allowing direct evaluation of SLC transporter activity.

Assay reliability
Mock HEK293 cells (Cat. No. GM1001) that do not overexpress the SLC transporter are available for use as a negative experimental control. Some compounds may demonstrate high background levels of transport due to the presence of endogenous transporters or non-specific binding.

Rapid results
The convenient product format enables data generation in 2 days from thawing of the cells to final results.

Related products
GM1002 Human OATP1B1 Transporter Cells
GM1004 Human OAT3 Transporter Cells
GM1005 Human OCT2 Transporter Cells
GM1006 Human OATP1B3 Transporter Cells
GM1008 Human OCT1 Transporter Cells
GM1013 Human NTCP Transporter Cells
GM1014 Human MATE1 Transporter Cells
GM1015 Human MATE2K Transporter Cells
GM1001 Mock Cells

Expi293F™ Inducible Cells (Gibco™)

The Expi293F Inducible cell line is derived from engineered Expi293F cells that stably express the tetracycline repressor (TetR) protein, thus enabling tetracycline-regulated protein expression. The cell line was established by the stable integration of the regulatory plasmid pcDNA6/TR, which encodes the TetR gene under the control of the human CMV promoter. It is a powerful tool for tetracycline-inducible high-yield expression of recombinant proteins.

Features of Expi293 Inducible cells include:
• Suspension, high-density culture in Expi293 Expression Medium
• Tetracycline-inducible protein expression with extremely low basal expression levels
• Equivalent protein yields to parental Expi293F cells—up to 1 g/L
• Significantly higher protein yields than other HEK293 Tet-inducible cell line alternatives

For inducible expression of your protein of interest, pcDNA5/TO mammalian expression vector (Cat. No. V103320) or an equivalent inducible vector is required for tetracycline-regulated gene expression.

Frozen cells are supplied in a vial containing 1 mL of cells at 1 x 107 viable cells/mL in Expi293 Expression Medium and 10% DMSO. The cells should be thawed directly into Expi293 Expression Medium.

293 H Cells, in CD 293 (Gibco™)

Gibco® 293-H cells were cloned from the original 293 cell line and adapted to Gibco® CD 293 Medium. The 293 cell line is a permanent line established from primary embryonic human kidney and transformed with sheared human adenovirus type 5 DNA. The E1A adenovirus gene is expressed in these cells and participates in transactivation of some viral promoters, allowing these cells to produce very high levels of protein. Gibco® 293-H cells in Gibco® CD 293 Medium feature:
• Quick thaw into suspension culture directly in Gibco® CD 293 Medium or Gibco® 293 SFM II
• Faster growth, good adherence, superior transfection efficiencies and high protein expression levels
• Documented lineage from a low passage Master Cell Bank
• Quality and performance testing

Quick thaw into suspension culture directly in Gibco® CD 293 Medium or Gibco® 293 SFM II
Each vial containing 7.5 × 106 cells can be thawed directly into suspension culture in either Gibco® CD 293 Medium or Gibco® 293 SFM II supplemented with 4 mM L-glutamine or GlutaMAX™ reagent. Protocols are provided in the product manual.

Faster growth, good adherence, superior transfection efficiencies and high protein expression levels
Gibco® 293-H cells were prepared from a clone selected for fast growth in serum-free medium, good adherence during plaque assays, superior transfection efficiencies and a high level of protein expression. Optimal transfection efficiencies are obtained with Lipofectamine™ 2000 or 293fectin™ transfection reagents.

Documented lineage from a low passage Master Cell Bank
The parental 293 cells were cloned by limiting dilution; the clone selected exhibited fast growth and high transfection efficiency. This clone was then re-cloned by limiting dilution; of these clones, the 293-H clone was then selected for its good adherence during plaque assays. The serum-free Master Cell Banks were prepared at approximately passage 25.

Quality and performance testing
Each lot of Gibco® 293 H cells is tested for cell growth and viability post-recovery from cryopreservation. In addition, the Master Seed Bank has been tested for contamination of bacteria, yeast, mycoplasma and virus and has been characterized by isozyme and karyotype analysis.

Product Use
For Research Use Only. Not for any animal or human therapeutic or diagnostic use. Caution: Handle as potentially biohazardous material under at least Biosafety Level 2 containment. This product contains Dimethyl Sulfoxide (DMSO), a hazardous material. Review the Material Safety Data Sheet before handling.

ExpiCHO-S™ Cells (cGMP Banked) (Gibco™)

ExpiCHO-S Cells are derived from a non-engineered sub-clone that was screened and isolated from CHO-S Chinese hamster ovary (CHO) cells (1). They are a core component of the ExpiCHO Expression System Kit. ExpiCHO-S Cells are maintained in suspension culture, have minimal tendency to clump, and grow to high density in ExpiCHO Expression Medium. As part of the ExpiCHO Expression System Kit, the cells generate superior protein yields by transient transfection compared to standard CHO, 293, and high-density Expi293F cells.

Features of ExpiCHO-S Cells (cGMP Banked) include:
• Derived from the same Master Seed Bank as our standard ExpiCHO-S Cells
• Fully documented cGMP-banked cell line
• A component of the fully integrated, animal origin-free ExpiCHO Expression System for applications requiring cGMP-banked cells
• Flexible commercial licensing options without royalties

Frozen cells are supplied in a vial containing approximately 1.1 mL of cells at 1 x 107 viable cells per mL in ExpiCHO Expression Medium and 10% DMSO. The cells should be thawed directly into ExpiCHO Expression Medium.

Fully documented cGMP cell line
After execution and payment for a Commercial Production License, a Cell Line Documentation Package (>200 pages and specific to the lot of cells purchased) is provided that includes the entire lineage history of the cells starting at receipt of the initial vial at Thermo Fisher Scientific through delivery to the purchaser, as well as all cell line characterization testing reports.

Commercial licensing that is simple, flexible, and without royalties
ExpiCHO-S Cells (cGMP banked) can be purchased directly from our catalog for research use only during internal development. As you move towards commercial use, a flexible Commercial Production or Service License structure is available without the burden of royalties, keeping it simple as a one-time payment. For more information regarding commercial licensing, email us at outlicensing@thermofisher.com.

Reference
1. Wurm, FM. CHO Quasispecies—Implications for Manufacturing Processes (2013) Processes 1:296-311.

Human NTCP SLC Transporter Cells (Gibco™)

TRANSiPORT Human NTCP SLC Transporter Cells are HEK293 cells that transiently overexpress the NTCP solute carrier (SLC) transport protein. Solute carriers are a super-family of membrane transporters that can affect pharmacokinetics and drug exposure by governing the transport of solutes in and out of cells. The NTCP SLC transporter protein is found in hepatocytes.

• Assess potential for transporter-mediated drug metabolism
• Easy to use format
• Obtain high quality results with a large signal-to-noise ratio

Choice of measurement system
Cell-based assays can be performed using radioisotope-labeled compounds, fluorescence-labeled compounds, or non-labeled compounds. The amount of substrate transported into the cells can be measured directly using a liquid scintillation counter, fluorescence plate reader, or LC-MS/MS, thereby allowing direct evaluation of SLC transporter activity.

Assay reliability
Mock HEK293 cells (Cat. No. GM1001) that do not overexpress the SLC transporter are available for use as a negative experimental control. Some compounds may demonstrate high background levels of transport due to the presence of endogenous transporters or non-specific binding.

Rapid results
The convenient product format enables data generation in 2 days from thawing of the cells to final results.

Related products
GM1002 Human OATP1B1 Transporter Cells
GM1003 Human OAT1 Transporter Cells
GM1004 Human OAT3 Transporter Cells
GM1005 Human OCT2 Transporter Cells
GM1006 Human OATP1B3 Transporter Cells
GM1008 Human OCT1 Transporter Cells
GM1014 Human MATE1 Transporter Cells
GM1015 Human MATE2K Transporter Cells
GM1001 Mock Cells