HepG2 Cells
































Essential facts on HepG2 cells
Description | HepG2 cells, a hepatoblastoma cell line, are a cornerstone in biological science, particularly in liver cancer research. The HepG2 cell line was first isolated in 1975 and initially misclassified as hepatocellular carcinoma, with the HepG2 cell line origin as hepatoblastoma being recognized later, clarifying years of scientific ambiguity. Human hepatic cell lines such as HepG2 are commonly used as in vitro models for primary human hepatocytes. These cell lines offer advantages such as indefinite proliferation, stable phenotype, easy accessibility, and ease of manipulation. However, they exhibit reduced expression of some metabolic functions compared to primary hepatocytes. Derived from hepatocellular carcinoma, HepG2 cells proliferate quickly and have an epithelial-like morphology, performing many specialized hepatic functions. Despite these differences, HepG2 cells are widely used in studying drug metabolism and toxicity, thanks to their resemblance to hepatocellular carcinoma and hepatoblastoma cells in terms of drug metabolism and transport proteins. HepG2 is a human liver cancer cell line often used in research, including studies on drug metabolism and toxicity. However, one of the limitations of hepatoma HepG2 cells is their altered expression of certain liver-specific functions, including the expression of cytochrome P450 enzymes. Cytochrome P450 enzymes are essential for the metabolism of xenobiotics (foreign compounds such as drugs and carcinogens) in the liver. The altered or reduced expression of these enzymes in HepG2 cells can affect their ability to accurately model the metabolism and elimination of xenobiotics, which is a critical aspect of liver function. The HepG2 cell line, alongside other hepatoma cell lines such as the Hep3B and human hepatoma HepaRG cell lines, contributes to a broader understanding of human liver carcinoma cells. The cell line stands out for its versatility, serving as an optimal choice for stable cell line generation, transfection studies, drug metabolism, and hepatotoxicity studies. Furthermore, the HepG2 cell line is pivotal in a range of applications, from 3D cell culture to high-throughput screening and toxicology. |
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Organism | Human |
Tissue | Liver |
Disease | Hepatocellular carcinoma |
Applications | This cell line is an optimal choice for transfection. Further, the HepG2 cells offer an array of applications, ranging from 3D cell culture and cancer research to high-throughput screening and toxicology. |
Synonyms | HEP-G2, Hep G2, HEP G2, Hep-G2, HEPG2 |
Characteristics of the HepG2 cell line
Age | 15 years |
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Gender | Male |
Ethnicity | Caucasian |
Morphology | Epithelial-like |
Growth properties | Adherent |
Identifiers / Biosafety / Citation
Citation | HepG2 (Cytion catalog number 300198) |
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Biosafety level | 1 |
Genomics of HepG2 human hepatoma cells
Receptors expressed | insulin, insulin-like growth factor II (IGF II) |
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Protein expression | p53 positive |
Tumorigenic | no |
Products | Albumin, alpha-fetoprotein (alpha fetoprotein), alpha1 acid glycoprotein (alpha-1 acid glycoprotein), alpha1 antitrypsin (alpha-1-antitrypsin), alpha1 antichymotrypsin, (alpha-1-antichymotrypsin), alpha2 HS glycoprotein (alpha-2-HS- glycoprotein), alpha2 macroglobulin (alpha-2-macroglobulin), beta lipoprotein (beta-lipoprotein), ceruloplasmin, C4 and C3 activator, fibrinogen, haptoglobin, plasminogen, retinol binding protein (retinolbinding protein), transferrin |
Karyotype | modal number = 55 (range = 50 to 60), has a rearranged chromosome 1 |
Handling of Hep G2 cells
Culture Medium | Ham's F12, w: 1.0 mM stable Glutamine, w: 1.0 mM Sodium pyruvate, w: 1.1 g/L NaHCO3 (Cytion article number 820600a) |
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Medium supplements | Supplement the medium with 10% FBS |
Passaging solution | Accutase |
Doubling time | 48 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 1:4 to 1:6 is recommended |
Seeding density | 2 to 3 x 10^4 cells/cm^2 during routine culture |
Fluid renewal | 2 to 3 times per week |
Freezing recovery | Start culture using the complete contents of the cryovial in 2xT25 cell culture flasks. The cells will recover within 48 to 72 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. |
Handling of cryopreserved cultures |
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Quality control / Genetic profile / HLA
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,y
CSF1PO: 10,11
D13S317: 9,13
D16S539: 12,13
D5S818: 11,13
D7S820: 10
TH01: 9
TPOX: 8,9
vWA: 17
D3S1358: 15,16
D21S11: 29,31
D18S51: 13,14
D8S1179: 15,16,17
FGA: 22,25
D2S1338: 19,2
D19S433: 15. Feb
|
HLA alleles |
A*: '02:01:01, '24:02:01
B*: '35:14:01, '51:08:01
C*: '04:01:01, '16:02:01
DRB1*: '13:02:01, '16:02:01
DQA1*: '01:02:01, '05:05:01
DQB1*: '03:01, '06:04
DPB1*: '02:01:02, '04:02:01
E: '01:01:01
|
Required products
One of its notable advantages is the ability to support cell growth without the need for serum supplementation. This eliminates potential interference caused by serum components, ensuring consistent and reliable experimental results. By providing a serum-free culture environment, Ham's F-12 Medium offers researchers greater control over their investigations.
Another key feature of Ham's F-12 Medium is its suitability for single-cell plating. This makes it an excellent choice for a variety of cell lines, including CHO cells, lung cells, and mouse L cells. The medium's optimized nutrient composition facilitates efficient attachment and growth of individual cells, enabling the establishment of homogeneous cell cultures with improved reproducibility.
Moreover, Ham's F-12 Medium has gained recognition as the preferred medium for the Clonal Toxicity Assay (CTA). This assay plays a critical role in assessing the cytotoxic effects of substances on cells. By utilizing Ham's F-12 Medium in the CTA, researchers can accurately evaluate the impact of various compounds or treatments on individual cells, providing valuable insights into toxicological profiles.
Quality control
pH = 7.2 +/
- 0.02 at 20-25°C.
Each lot has been tested for sterility and absence of mycoplasma and bacteria.
Maintenance
Keep refrigerated at +2°C to +8°C in the dark. Freezing and warming up to +37° C minimize the quality of the product.
Do not heat the medium to more than 37° C or use uncontrollable sources of heat (e.g., microwave appliances).
If only a part of the medium is to be used, remove this amount from the bottle and warm it up at room temperature.
Shelf life for any medium except for the basic medium is 8 weeks from the date of manufacture.
Composition
Components
mg/L
Inorganic Salts
Calcium chloride x 2H2O
44,00
Copper(II) sulfate x 5H2O
0,00
Iron (II) sulfate x 7H2O
0,83
Magnesium chloride x 6H2O
122,00
Potassium chloride
223,65
Sodium chloride
7599,00
di-Sodium hydrogen phosphateanhydrous
142,04
Zinc sulfate x 7H2O
0,86
Other Components
D(+)-Glucose anhydrous
1801,60
Hypoxanthine
4,08
Linoleic acid
0,08
DL-α-Lipoic acid
0,21
Phenol red
1,20
Putrescine x 2HCl
0,16
Sodium pyruvate
110,00
Thymidine
0,73
NaHCO3
1176,00
Amino Acids
L-Alanine
8,91
L-Arginine x HCl
210,70
L-Asparagine x H2O
15,01
L-Aspartic acid
13,31
L-Cysteine x HCl x H2O
35,12
L-Alanyl-L-Glutamine
217,30
L-Glutamic acid
14,71
Glycine
7,51
L-Histidine x HCl x H2O
20,96
L-Isoleucine
3,94
L-Leucine
13,12
L-Lysine x HCl
36,54
L-Methionine
4,48
L-Phenylalanine
4,96
L-Proline
34,53
L-Serine
10,51
L-Threonine
11,91
L-Tryptophan
2,04
L-Tyrosine
5,44
L-Valine
11,71
Vitamins
D(+)-Biotin
0,01
D-Calcium pantothenate
0,24
Choline chloride
13,96
Folic acid
1,32
myo-Inositol
18,02
Nicotinamide
0,04
Pyridoxine x HCl
0,06
Riboflavin
0,04
Thiamine x HCl
0,34
Vitamin B12
1,36
- 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
Certificate of Analysis (CoA)
Lot Number | Certificate Type | Date | Catalog Number |
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300198-240624 | Certificate of Analysis | 23. May. 2025 | 300198 |
300198-260523 | Certificate of Analysis | 23. May. 2025 | 300198 |