HEK293T Cells
























Essential facts about HEK293T cells
Description | HEK 293T, a highly transfectable derivative of the parental HEK 293 cell, stands out as a versatile and powerful tool in the field of biotechnology for the production of recombinant proteins and various types of vaccines. HEK-293T cells were generated by transfecting embryonic kidney 293 cells with a plasmid encoding the SV40 large T antigen. The original HEK293 cell line was developed from the epithelial cells of human embryonic kidney tissue, with its transformation occurring in what was notably the 293rd experiment conducted by the researchers. In the realm of vaccine development, the 293T embryonic kidney cells are pivotal for viral vector production, including adenovirus vectors. HEK293T cells, under specific culture conditions, are transfected with vectors carrying adenoviral and retroviral elements, including the SV40 origin of replication, leading to the production of virus-like particles (VLPs). The VLPs, devoid of viral genetic material, are key in forming the basis of subunit and VLP-based vaccines. The recombinant protein production in 293T cells is facilitated by various transfection methods, with an emphasis on the generation of AP fusion proteins and other protein types that form the antigenic component of vaccines. The 293T cell line's genome engineering capabilities allow for the customization of expression constructs, further boosting the production of viral vectors. This, coupled with the ability to produce proteins in suspension culture or adherent conditions, makes the 293T cell line a full-stack solution for modern vaccine development. |
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Organism | Human |
Tissue | Kidney |
Applications | Vaccine development |
Synonyms | Hek293T, HEK-293T, HEK 293T, HEK-293-T, HEK 293 T, 293-T, 293 T, 293T, Human Embryonic Kidney 293T, 293tsA1609neo |
Details about HEK293
Age | Fetus |
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Gender | Female |
Morphology | Epithelial-like |
Growth properties | Adherent |
Documentation
Citation | HEK293T (Cytion catalog number 300189) |
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Biosafety level | 1 |
NCBI_TaxID | 9606 |
CellosaurusAccession | CVCL_0063 |
Genetic profile
Receptors expressed | Vitronectin |
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Protein expression | CEA negative, p53 positive |
Tumorigenic | In nude mice |
Handling HEK 293T cells
Culture Medium | EMEM (MEM Eagle), w: 2 mM L-Glutamine, w: 2.2 g/L NaHCO3, w: EBSS (Cytion article number 820100a) |
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Supplements | Supplement the medium with 10% FBS and 1% NEAA |
Dissociation Reagent | Accutase |
Doubling time | 30 hours |
Subculturing | Remove the old medium from the adherent cells and wash them with PBS that lacks calcium and magnesium. For T25 flasks, use 3-5 ml of PBS, and for T75 flasks, use 5-10 ml. Then, cover the cells completely with Accutase, using 1-2 ml for T25 flasks and 2.5 ml for T75 flasks. Let the cells incubate at room temperature for 8-10 minutes to detach them. After incubation, gently mix the cells with 10 ml of medium to resuspend them, then centrifuge at 300xg for 3 minutes. Discard the supernatant, resuspend the cells in fresh medium, and transfer them into new flasks that already contain fresh medium. |
Split ratio | A ratio of 1:3 to 1:4 is recommended |
Seeding density | 1 x 10^4 cells/cm^2 will yield in a confluent layer in about 4 days |
Fluid renewal | 2 times per week |
Post-Thaw Recovery | After thawing, plate the cells at 5 x 10^4 cells/cm^2 and allow the cells to recover from the freezing process and to adhere for at least 24 hours. |
Freeze medium | As a cryopreservation medium, use complete growth medium (including FBS) + 10% DMSO for adequate post-thaw viability, or CM-1 (Cytion catalog number 800100), which includes optimized osmoprotectants and metabolic stabilizers to enhance recovery and reduce cryo-induced stress. |
Thawing and Culturing Cells |
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HEK cell quality verification
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. |
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STR profile |
Amelogenin: x,x
CSF1PO: 11,12
D13S317: 12,13,14
D16S539: 9,13
D5S818: 8,9
D7S820: 11
TH01: 7,9.3
TPOX: 11
vWA: 16,18,19,20
D3S1358: 15,16,17,18
D21S11: 28,30.2
D18S51: 17,18
Penta E: 7,15
Penta D: 9,1
D8S1179: 11,12,13,14
FGA: 22,23
D2S1338: 19
D19S433: 18
PEZ6: EB1
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Required products
What's in EMEM?
EMEM is a modified version of Eagle's minimum essential medium, containing Earle's Balanced Salt Solution, non-essential amino acids, L-glutamine, sodium pyruvate, and sodium bicarbonate. It's important to note that this level of sodium bicarbonate is intended for use in 5% CO2 in the air. To maintain its effectiveness, storing the medium at 2°C to 8°C in the dark when not in use is recommended.
What is EMEM used for?
Eagle's minimal essential medium (EMEM) is a cell culture medium that can maintain cells in tissue culture. The medium contains higher concentrations of amino acids, allowing for a more accurate approximation of the protein composition of cultured mammalian cells. EMEM may be used to cultivate various cells, including fibroblasts, human liver cancer cell line (HepG2) cells and human fetal brain progenitor-derived astrocyte cells (PDA). It is typically used in the presence of fetal bovine serum (FBS), calf, or horse sera.
How is EMEM different from other cell culture media?
While EMEM and Dulbecco's modified Eagle's medium (DMEM) share some similarities, they also differ. Both media lack protein and contain the amino acids, salts, glucose, and vitamins required to provide a cell with energy and maintain it in tissue culture. However, the DMEM formulation is modified to contain up to four times more vitamins and amino acids and two to four times more glucose than EMEM. It's worth noting that EMEM is also different from the original MEM formulation.
Quality Control
Sterile-filtered
Storage and Shelf Life
Store at +2°C to +8°C, protected from light.
Once opened, store at 4°C and use within 6–8 weeks.
Shipping Conditions
Ambient temperature
Maintenance
Keep refrigerated at +2°C to +8°C in the dark. Avoid freezing and frequent warming to +37°C, as it reduces product quality.
Do not heat the medium beyond 37°C or use uncontrolled heat sources such as microwave appliances.
If only part of the medium is to be used, remove the required amount and warm it to room temperature before use.
Composition
Category
Components
Concentration (mg/L)
Amino Acids
L-Arginine HCl
126.00
L-Cystine 2 HCl
31.30
L-Glutamine
292.00
L-Histidine HCl H2O
42.00
L-Isoleucine
52.00
L-Leucine
52.00
L-Lysine HCl
72.50
L-Methionine
15.00
L-Phenylalanine
32.00
L-Threonine
48.00
L-Tryptophan
10.00
L-Tyrosine 2 Na 2 H2O
51.90
L-Valine
46.00
Vitamins
Choline Chloride
1.00
Vitamins
D-Calcium Pantothenate
1.00
Folic Acid
1.00
myo-Inositol
2.00
Nicotinamide
1.00
Pyridoxal HCl
1.00
Riboflavin
0.10
Thiamine HCl
1.00
Inorganic Salts
CaCl2 2 H2O
265.00
Inorganic Salts
KCl
400.00
MgSO4
97.67
NaCl
6800.00
NaHCO3
2200.00
NaH2PO4
122.00
Other Components
D-Glucose
1000.00
Other Components
Phenol Red Sodium Salt
11.00
- A Gentle Alternative to Trypsin
Accutase is a cell detachment solution that is revolutionizing the cell culture industry. It is a mix of proteolytic and collagenolytic enzymes that mimics the action of trypsin and collagenase. Unlike trypsin, Accutase does not contain any mammalian or bacterial components and is much gentler on cells, making it an ideal solution for the routine detachment of cells from standard tissue culture plasticware and adhesion coated plasticware. In this blog post, we will explore the benefits and uses of Accutase and how it is changing the game in cell culture.
Advantages of Accutase
Accutase has several advantages over traditional trypsin solutions. Firstly, it can be used whenever gentle and efficient detachment of any adherent cell line is needed, making it a direct replacement for trypsin. Secondly, Accutase works extremely well on embryonic and neuronal stem cells, and it has been shown to maintain the viability of these cells after passaging. Thirdly, Accutase preserves most epitopes for subsequent flow cytometry analysis, making it ideal for cell surface marker analysis.
Additionally, Accutase does not need to be neutralized when passaging adherent cells. The addition of more media after the cells are split dilutes Accutase so it is no longer able to detach cells. This eliminates the need for an inactivation step and saves time for cell culture technicians. Finally, Accutase does not need to be aliquoted, and a bottle is stable in the refrigerator for 2 months.
Applications of Accutase
Accutase is a direct replacement for trypsin solution and can be used for the passaging of cell lines. Additionally, Accutase performs well when detaching cells for the analysis of many cell surface markers using flow cytometry and for cell sorting. Other downstream applications of Accutase treatment include analysis of cell surface markers, virus growth assay, cell proliferation, tumor cell migration assays, routine cell passage, production scale-up (bioreactor), and flow cytometry.
Composition of Accutase
Accutase contains no mammalian or bacterial components and is a natural enzyme mixture with proteolytic and collagenolytic enzyme activity. It is formulated at a much lower concentration than trypsin and collagenase, making it less toxic and gentler, but just as effective.
Efficiency of Accutase
Accutase has been shown to be efficient in detaching primary and stem cells and maintaining high cell viability compared to animal origin enzymes such as trypsin. 100% of cells are recovered after 10 minutes, and there is no harm in leaving cells in Accutase for up to 45 minutes, thanks to autodigestion of Accutase.
In summary
In conclusion, Accutase is a powerful solution that is changing the game in cell culture. With its gentle nature, efficiency, and versatility, Accutase is the ideal alternative to trypsin. If you are looking for a reliable and efficient solution for cell detachment, Accutase is the solution for you.
Phosphate-buffered saline (PBS) is a widely used buffer solution in biological and chemical research. It plays a crucial role in maintaining the pH balance and osmolarity during various experimental procedures, including tissue processing and cell culture. Our PBS solution is meticulously formulated with high-purity ingredients to ensure stability and reliability in every experiment. The osmolarity and ion concentrations of our PBS closely mimic those of the human body, making it isotonic and non-toxic to most cells.
Composition of Our PBS Solution
Our PBS solution is a pH-adjusted blend of ultrapure-grade phosphate buffers and saline solutions. At a 1X working concentration, it contains:
8000 mg/L Sodium chloride (NaCl)
200 mg/L Potassium chloride (KCl)
1150 mg/L Sodium phosphate dibasic anhydrous (Na2HPO4)
200 mg/L Potassium phosphate monobasic anhydrous (KH2PO4)
This composition ensures an optimal pH and ionic balance, suitable for a wide range of biological applications.
Applications of Our PBS Solution
Our PBS solution is ideal for various applications in biological research. Its isotonic and non-toxic properties make it suitable for substance dilution and cell container rinsing. PBS solutions containing EDTA are effective for disengaging attached and clumped cells. However, divalent metals such as zinc should not be added to PBS, as this can cause precipitation. In such cases, Good's buffers are recommended. Additionally, our PBS solution is an acceptable alternative to viral transport medium for the transport and storage of RNA viruses, including SARS-CoV-2.
Quality Control
Sterile-filtered
Storage and Shelf Life
Store at +2°C to +25°C, protected from light.
Once opened, store at 2°C to 25°C and use within 24 months.
Shipping Conditions
Ambient temperature
Maintenance
Keep refrigerated at +2°C to +8°C in the dark. Avoid freezing and frequent warming to +37°C, as it reduces product quality.
Do not heat the medium beyond 37°C or use uncontrolled heat sources such as microwave appliances.
If only part of the medium is to be used, remove the required amount and warm it to room temperature before use.
Composition
Category
Components
Concentration (mg/L)
Salts
Potassium chloride
200
Potassium phosphate monobasic anhydrous
200
Sodium chloride
8000
Sodium phosphate dibasic anhydrous
1150
Related products
Certificate of Analysis (CoA)
Lot Number | Certificate Type | Date | Catalog Number |
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300189-160824 | Certificate of Analysis | 26. May. 2025 | 300189 |
300189-050824 | Certificate of Analysis | 23. May. 2025 | 300189 |
Material Transfer Agreement
If you intend to use Cytion cell lines solely for internal research at a single research site, please complete and sign our Material Transfer Agreement (MTA) and submit it along with your order.
For any commercial applications - including but not limited to fee-for-service work, quality control testing, product release, diagnostic use, or regulatory studies - please complete the Intended Use Form so we can prepare a suitable agreement tailored to your project.
Please note: The MTA applies only to certain cell lines. If this notice and the MTA document appear on a product page, the agreement is applicable. For cell lines not covered by the MTA, no reference to the agreement will be shown. The MTA is not valid for customers in the Americas, China, or Taiwan. Please contact our U.S. entity to receive the appropriate agreement.