The HEK293 suspension-adapted cell line is a variant of the human embryonic kidney 293 (HEK293) cells that has been modified to grow in suspension culture rather than adherent culture. This adaptation is significant for industrial applications where large-scale protein production is required. The cells maintain many of the characteristics of the original HEK293 line, including a robust transient transfection efficiency and the ability to post-translationally modify expressed proteins in a manner similar to that of native human cells.
The HEK293 suspension-adapted cell line is a variant of the human embryonic kidney 293 (HEK293) cells that has been modified to grow in suspension culture rather than adherent culture. This adaptation is significant for industrial applications where large-scale protein production is required. The cells maintain many of the characteristics of the original HEK293 line, including a robust transient transfection efficiency and the ability to post-translationally modify expressed proteins in a manner similar to that of native human cells.
These cells are particularly valued in the biotechnology and pharmaceutical industries for the production of recombinant proteins and viruses for gene therapy and vaccine development. The adaptation to suspension culture allows for easier scalability and simplifies the harvesting process, making it more suitable for commercial-scale bioprocessing. The HEK293 suspension-adapted cell line supports various viral production systems, including adenovirus, lentivirus, and adeno-associated virus (AAV), which are pivotal in therapeutic applications and research.
Overall, the HEK293 suspension-adapted cell line is a crucial tool in the fields of molecular biology and bioprocessing, providing a versatile platform for the production of various biologically active molecules. Its ease of genetic manipulation and ability to produce proteins that are correctly folded and post-translationally modified according to human cell patterns make it an indispensable resource in many advanced therapeutic and research settings.
Gender
Female
Age
Fetus
Growth Properties
Suspension
Tissue
Kidney
Biosafety level
1
Citation
HEK293 suspension-adapted (Cytion catalog number 300686)
Protein Expression
CEA negative, p53 positive
Tumorigenic
In nude mice
Virus Susceptibility
Transformed with adenovirus 5 DNA adenovirus 5 DNA
Receptors Expressed
Vitronectin
Culture Medium
Panserin 293S (from PanBiotech)
Freeze Medium
CM-1 (Cytion catalog number 800100) or CM-ACF (Cytion catalog number 806100)
Handling of Cryopreserved Cultures
1. Confirm that the vial remains deeply frozen upon delivery, as cells are shipped on dry ice to maintain optimal temperatures during transit.
2. Upon receipt, either store the cryovial immediately at temperatures below -150°C to ensure the preservation of cellular integrity, or proceed to step 3 if immediate culturing is required.
3. For immediate culturing, swiftly thaw the vial by immersing it in a 37°C water bath with clean water and an antimicrobial agent, agitating gently for 40-60 seconds until a small ice clump remains.
4. Perform all subsequent steps under sterile conditions in a flow hood, disinfecting the cryovial with 70% ethanol before opening.
5. Carefully open the disinfected vial and transfer the cell suspension into a 15 ml centrifuge tube containing 8 ml of room-temperature culture medium, mixing gently.
6. Centrifuge the mixture at 300 x g for 3 minutes to separate the cells and carefully discard the supernatant containing residual freezing medium. Optionally, skip centrifugation but remove any remaining freezing medium after 24 hours.
7. Gently resuspend the cell pellet in 10 ml of fresh culture medium. For adherent cells, divide the suspension between two T25 culture flasks; for suspension cultures, transfer all the medium into one T25 flask to promote effective cell interaction and growth.
8. Adhere to established subculture protocols for continued growth and maintenance of the cell line, ensuring reliable experimental outcomes.
Subculturing
Maintain cultures by periodically adding or replacing the medium. Initiate cultures with a density of 2 x 10^5 cells/ml and keep the cell concentration within the range of 1 x 10^5 to 1 x 10^6 cells/ml for optimal growth.
Seeding Density
3 x 10^5 cells/ml
Fluid Renewal
2 to 3 times per week
Sterility
Mycoplasma contamination is excluded using both PCR-based assays and luminescence-based mycoplasma detection methods.
To ensure there is no bacterial, fungal, or yeast contamination, cell cultures are subjected to daily visual inspections.
This cell line is available in the U.S. For non-profit users, please sign and return the Supply Agreement to [email protected] when placing an order. For commercial users, please complete the CLEAR Form as additional usage fee may apply based on the given intended usage. For further details, please contact us at [email protected].
What products or services do you offer? We offer high-quality cell lines including human cells, animal cell lines, fluorescence-tagged cell lines, iPSC lines, as well as different human primary cells (human mesenchymal stem cells, fibroblasts, follicular dendritic cells, HUVEC, oral cells). In addition, you can acquire more than 30 different serum-containing media and 10 serum-free media for the cultivation and maintenance of various cell types. Our service includes viral testing, mycoplasma detection service, cell line authentication, cell banking and species identification. Moreover, we offer bulk and custom manufacturing. Where can I find references from other researchers that have used your products? If available, you will find the references with the respective product. How to determine the concentration of cells in a suspension? Multiply the average cell count from each of the hemocytometer's 16 corner squares by 10,000. Final viability counts per milliliter of sample are obtained by multiplying the initial count by 5, to account for the 1:5 dilution caused by the addition of trypan blue. Example: (50 + 40 + 45 +52) ÷ 4 = 46.75 75 x 10,000 (10^4) = 467,500 467,500 x 5 = 2,337,500 live cells/mL in original cell suspension Alternatively use an automated cell counting device such as the CellDrop from DeNovix or the NucleoCounter NC-200 from Chemometec. How to store the purchased cells? We strongly recommend to store the cells after receipt on dry ice for a very short time (1 day at the most) at -80°C and start culturing as soon as possible. If immediate culturing is not possible, the cryovial(s) must be stored at temperatures below -150°C. The quality of the permanent cell lines should be controlled within 60 days after receipt. Approach: Thaw the cells Determine cell viability Culture the cells Make up your own master stock. How to store the purchased media and supplements? All media available in our shop must be stored at 4 °C in the refrigerator. The supplements available have different storage conditions. Please refer to the information on the data sheets supplied. How do I establish a culture from cryopreserved cells? Bring water to a temperature of 37 °C in a beaker. Remove a vial of cells from liquid nitrogen storage with care, making sure to shield your hands and eyes. Loosen the vial's cap for 10 seconds to allow any liquid nitrogen to escape the threads, and then re-tighten the top. To thaw, just submerge the vial's lower half in the 37 °C water for four minutes. Wipe the outside of the vial with disinfectant solution and place it in a laminar flow culture hood of Class II, type A, once the contents have thawed until just a little piece of ice remains. To disperse the cells, open the vial and pipette the suspension up and down with a 1 mL pipette. Take out 20 µl, then add 20 µl of trypan blue solution to the cell suspension (e.g., Cat. No. 15250-061). Use a hemocytometer or an automatized cell count system to calculate the number of viable cells per milliter. Dilute the vial's contents (1 mL) to the concentration suggested in the product instructions. After inoculation, agitate the medium in the flasks to disperse the cells. Numerous cell types adhere rapidly to culture surfaces; If the medium is not evenly distributed immediately after inoculation, the cells may grow in an uneven pattern. Maintain the cultures in a cell culture incubator at 37 degrees Celsius with 5% carbon dioxide and 95% air humidity. Ideally, the culture should be left alone for at least 24 hours after it has been started. How do I know what kind of cell culture medium i need to use for my specific cell line? The information on the cell culture medium we do recommend for a specific cell line is given on the product page for the cell line as well as in the product information. The most common cell culture media are available as basic as well as ready-to-use (containing e.g., FBS and other supplements) formulations. Some cell lines such as NFS-60 or TF-1 need special ingredients in their culture medium. Our Conditioned media are a good supply of these factors. In order to cryopreserve the cells in liquid nitrogen, we recommend to use either CM-1, which contains serum, or the serum free freeze medium CM-ACF. What are living or growing cultures? Our product range also includes living cell cultures. Depending on the cell type, these are adherent or suspended cells. How often should I change the media? After thawing and plating cryopreserved cells, the first medium change should be performed after 24 hours or overnight to remove any remaining DMSO and dead cells. The medium should then be replaced every 24-48 hours until the cells are ready to be passed. Protocols for each cell line contain more specific instructions. Can I expand your cells and re-freeze them? Our cryopreserved cell lines or growing cultures may be expanded and cryopreserved again. However, the process of cryopreservation may alter the growth performance of cells. Do I need to spin the cells out of the cryopreservation medium to plate them? We do not advise spinning cells out of cryopreservation medium before plating. When centrifugation is performed at too high speeds, it might cause damage to cells. In our cell culture labs, we have found that at low concentrations, DMSO is safe for the cells. We advise diluting the cells in culture medium until the final DMSO content is less than 0.4% (v/v). Do I need a surface matrix to culture my cells in 2D? For the most of our cells you do not need a special surface matrix for 2D culture. Only our induced pluripotent stem cells (hiPSCs) require a culture surface substrate that maintains pluripotency. Here we recommend to use iMatrix or Matrigel. How can I dissociate my cells from the cell culture plates? For the detachment of most of the cells provided by CLS (except for e.g., for some keratinocyte cells such as HaCaT), we recommend to use Accutase which gently detaches the cells from the bottom of the cell culture flask. Accutase works with most cell lines. In case of the need of alternative detachment solutions, these are indicated in the specific product information of the cell line. Answers to frequently asked questions on cell lines, strains, and types When cells are separated from a tissue to generate a primary culture, a confluent monolayer or dense cell suspension is formed due to the cells' ability to proliferate in vitro. In accordance with the traditional concept, a cell line is created after the initial collection and subculturing of this population (Freshney, R.I. (1987). Methods used to grow new tissues from animal cells. (New York: Alan R. Liss, Inc.) A primer on the fundamentals. Over the course of a cell line's existence, those cells with the greatest growth potential will prevail, leading to some genetic and behavioral homogeneity. With this terminology, a population of cells that has undergone a genetic alteration that allows for infinite growth is called a continuous cell line. Constant cell lines are usually aneuploid. Even while it is possible to culture continuous cell lines through a large number of subcultures, there is a risk that additional genotypic and, by extension, phenotypic changes will occur at these very high passage numbers. One can achieve immortality in a variety of ways, including naturally or with the help of viruses or chemicals. Understand that different research teams may use slightly different operational definitions of these concepts. Many researchers avoid calling a population a "cell line" until they can prove that it has undergone some sort of genetic mutation. Positively selected from a culture, either by cloning or some other means, a small subset of cells is called a cell strain. Since its original parent lineage was established, most cell strains have experienced significant genetic modifications. Some subsets of cells may have become more tumorigenic than the original line, or the transfection process may have resulted in the creation of a new strain. Cell types, which include keratinocytes and melanocytes, are collections of cells that have a common phenotype. That's why it's safe to say that every donor's keratinocytes are the same cell type. What are stem cells? Stem cells aid in the production of new cells in healthy tissues and may aid in the repair of injured or damaged tissues. They serve as the foundation for the distinct cell types that comprise each organ in the body. Stem cells are distinguished from other cells by their ability to self-renew, proliferate for an extended period of time, and, under specific conditions, differentiate into specialized cells with distinct functions (phenotypes), including but not limited to cardiac cells, liver cells, fat cells, bone cells, cartilage cells, nerve cells, and connective tissue cells. Multipotency is the capacity of cells to differentiate into a variety of other cell types. What scientists discover regarding the regulation of stem cell differentiation can serve as the foundation for the development of novel treatments for a variety of life-threatening diseases and injuries. What are induced pluripotent stem cells? Somatic (adult, non-germline) cells that have been reverted to an embryonic stem cell-like state give rise to induced pluripotent cells. As with embryonic stem cells, iPS cells are pluripotent because they can differentiate into any cell in the body. The process of creating these cells, commonly referred to as "reprogramming," involves the introduction of three to four genes for transcription factors into a somatic cell via retroviruses. Recent techniques have replaced and reduced the number of genes required for transformation, utilized alternative gene delivery methods, or sought to replace genes with chemical factors. Patients with diseases such as ALS, Parkinson's, or cardiovascular disease can have their cells induced to form iPS cells. When differentiated into more specialized cell types, iPS cells have multiple applications, including the development of assays for studying disease processes, screening drug candidates for safety and efficacy, and use in regenerative medicine. What is the shelf life of your culture media? You can find the information about shelf life of our products in the material safety data sheets at the respective product as pdf-file under product information/downloads. Is it possible to freeze and thaw your culture media and supplements? It is not recommended to store our cell culture media products under 4 °C. Regarding our supplement products please use the individual safety data sheets to get information’s about storing.
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