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Drosophila S2 Cells in Schneider's Medium (Gibco™)

Gibco® Drosophila S2 cells are used for heterologous protein expression in the DrosophilaM Expression System (DES®). The S2 cell line was derived from a primary culture of late stage (20-24 hours old) Drosophila melanogaster embryos. Many features of the S2 cell line suggest that it is derived from a macrophage-like lineage. S2 cells grow at room temperature in ambient CO2 as a loose, semi-adherent monolayer in tissue culture flasks and in suspension in spinners and shake flasks. Drosophila S2 cells (frozen in Gibco® Schneider's Medium) feature:
• Expression of protein transiently or through generation of stable cell lines
• Quality and performance testing

Expression of protein transiently or through generation of stable cell lines
Drosophila S2 cells can be transfected with the recombinant expression vector alone for transient expression studies or in combination with a selection vector (e.g. pCoHygro or pCoBlast) to generate stable cell lines. A variety of DES® kits are available to allow expression of the recombinant protein of interest in S2 cells.

Quality and performance testing
Each lot of Gibco® S2 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.

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.

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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293FT Cell Line (Invitrogen™)

The 293FT Cell line is ideal for generating high-titer lentivirus using the ViraPower™ Lentiviral Expression System. The 293FT Cell Line is a fast-growing, highly transfectable clonal isolate derived from human embryonal kidney cells transformed with the SV40 large T antigen. When a ViraPower™ expression vector and the ViraPower™ Lentiviral Packaging Mix are co-transfected into 293FT cells, high levels of the viral RNA and the gag⁄pol and rev proteins required for packaging are produced.

Advantages
• Fast growing
• Highly transfectable
• Produces high viral titers
• Allows very high levels of protein

Key Features
• Presence of the SV40 large T antigen allows very high levels of protein to be expressed from vectors containing the SV40 origin. Higher, in fact, than other 293 cells or the T antigen-transformed COS cells classically used.

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

HepaRG™ Cells, Cryopreserved (Gibco™)

The HepaRG™ cell line is an immortalized hepatic cell line that retains many characteristics of primary human hepatocytes. HepaRG™ cells are terminally differentiated and provided in a convenient cryopreserved format. For scientists who need reproducible metabolism data, HepaRG™ cells are an in vitro tool that provides reproducible results in a metabolically complete and scalable system.
• Obtain biologically relevant results from a metabolically complete system
• Assess the drug-drug interaction potential of your compound
• Experience reproducible results from a single population of cells

Biologically Relevant
HepaRG™ cells exhibit many characteristics of primary human hepatocytes including morphology, expression of key metabolic enzymes, expression of nuclear receptors, and drug transporters. Unlike HepG2 and Fa2N-4 cells, HepaRG™ cells have high P450 activity and complete expression of all nuclear receptors.

Predict Metabolism-based Drug-drug Interaction
HepaRG™ cells respond to prototypical P450 inducers and inhibitors to the same extent as primary hepatocytes, allowing HepaRG™ cells to be used to evaluate potential drug-drug interactions.

Infinitely Reproducible
HepaRG™ cells exhibit many characteristics of primary human hepatocytes and are essentially a single donor. These features allow users to obtain physiological relevant results for metabolism-based drug-drug interactions without the concern of donor variability and limited lot sizes that come with relying on donor tissue. (Note: The cells we provide are terminally differentiated).

HepaRG™ is a trademark of BioPredic International.

KM71H, Pichia pastoris Yeast Strain (Invitrogen™)

Several Pichia pastoris strains are available to allow you to optimize expression and recovery of your protein of interest. Table 1 provides information to help you choose the appropriate strain.

Flp-In™ T-REx™ 293 Cell Line (Invitrogen™)

Flp-In™ 293 T-REx cell lines is designed for rapid generation of stable cell lines that ensure homogenous expression of your 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. 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.

The Flp-In™-293 work well with Flp-In™ vectors that express a gene from the CMV promoter (e.g., pcDNA™5⁄FRT, pcDNA5⁄FRT⁄V5-His-TOPO®, and pSecTag⁄FRT⁄V5-His-TOPO®).

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.

Mimic™ Sf9 Insect Cells (Gibco™)

Mimic™ Sf9 Insect Cells are a derivative of the Sf9 insect cell line. These cells have been modified to stably express a variety of mammalian glycosyltransferases. Typically, insect cells are unable to process N-glycans to the extent that mammalian cells do. This can affect protein structure, function, antigenicity, and enzymatic activity. The addition of mammalian glycosyltransferases to the Mimic™ Sf9 Insect Cells allows for production of biantennary, terminally sialyated N-glycans from insect cells (1). The cells can be used to produce more mammalian-like proteins in both baculovirus and stable insect expression systems.

1 ml of Mimic™ Sf9 Insect Cells are provided frozen in 60% complete TNM-FH, 30% FBS, 10% DMSO at a concentration of 107 cells/ml. Store frozen Mimic™ Sf9 Insect Cells in liquid nitrogen. Cells are guaranteed stable for 6 months when properly stored.

ExpiCHO-S™ Cells (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 20 passages after thaw
• Maintain high cell viability and productivity for greater than 10 days post-transfection as part of the ExpiCHO expression system
• Also available from a cGMP bank (Cat. No. A37785)

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.

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.

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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GM1013 Human NTCP Transporter Cells
GM1014 Human MATE1 Transporter Cells
GM1015 Human MATE2K Transporter Cells
GM1001 Mock Cells

Flp-In™-293 Cell Line (Invitrogen™)

Flp-In™ 293 cell line is designed for rapid generation of stable cell lines that ensure high level expression of your 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. Flp-In™-293 cell line was created by transfecting the parent cell lines with pFRT⁄lacZeo and selecting for stable Zeocin™- resistant clones.

Flp-In™-293 cells work well with Flp-In™ vectors that express a gene from the CMV promoter (e.g., pcDNA™5⁄FRT, pcDNA5⁄FRT⁄V5-His-TOPO®, and pSecTag⁄FRT⁄V5-His-TOPO

SMD1168H, Pichia pastoris Yeast Strain (Invitrogen™)

Several Pichia pastoris strains are available to allow you to optimize expression and recovery of your protein of interest. Table 1 provides information to help you choose the appropriate strain.

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.

FreeStyle™ 293-F Cells (Gibco™)

The FreeStyle™ 293-F cell line is derived from the 293 cell line and is intended for use with the FreeStyle™ MAX 293 Expression System or FreeStyle™ 293 Expression System. FreeStyle™ 293-F cells are adapted to suspension culture in FreeStyle™ 293 Expression Medium. Frozen cells are supplied in and may be thawed directly into FreeStyle™ 293 Expression Medium. The FreeStyle™ 293-F cell line is supplied in a vial containing 1 mL of cells at 1 x 107 viable cells/mL in 90% FreeStyle™ 293 Expression Medium and 10% DMSO. Characteristics of FreeStyle™ 293-F cells:

• Prepared from low-passage Master Cell Bank cultures derived from parental 293-F cells that were re-cloned by limiting dilution. The 293 clonal derived cultures are maintained in serum-free conditions for only 30 to 35 total passages.
• Adapted to high-density, serum-free, suspension growth and may be maintained in FreeStyle™ 293 Expression Medium.
• Demonstrate high transfection efficiencies with FreeStyle™ MAX reagent.
• Suspension cultures may be transfected in FreeStyle™ 293 Expression Medium without the need to change media.
• Permits transfection of cells at large volumes.