HEp-2 Cells
























Introduction to Hep-2 cells
Description | The HEp-2 cell line, originally believed to be derived from laryngeal cancer cells, was later identified through DNA fingerprinting and the presence of HeLa marker chromosomes as being contaminated with HeLa cells, a cell line that was derived from cervical cancer. Despite this, the HEp-2 cell line remains extensively utilized in indirect immunofluorescence to detect antinuclear antibodies (ANAs), which are key in diagnosing conditions like systemic lupus erythematosus and systemic sclerosis. The indirect immunofluorescence assay (IIFA) using HEp-2 cells, which provides clear positive or negative results, is the standard method for testing antinuclear antibodies. This straightforward approach is crucial for diagnosing and classifying different systemic autoimmune diseases. The patterns of autoantibodies observed in indirect immunofluorescence on HEp-2 cells, especially in the context of rheumatology, provide invaluable insights into various rheumatic diseases. Furthermore, the comprehensive review of antigens expressed by HEp-2 human cells under different culture conditions enables the identification of specific ANAs linked to diseases like lupus. In conclusion, while the contamination of cell lines like HEp-2 with HeLa cells has prompted concerns in cancer research about the accuracy and reliability of results and their clinical relevance, the utility of Hep-2 in the detection of antinuclear antibodies and its application across various research disciplines underscore its continued importance. The HEp-2 cell line serves as an essential tool in diagnosing and classifying autoimmune diseases, among other applications. |
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Organism | Human |
Tissue | Larynx |
Disease | Adenocarcinoma |
Applications | In rheumatology, indirect immunofluorescence using HEp-2 cells plays a crucial role in diagnosing autoimmune diseases, including systemic lupus erythematosus and systemic sclerosis |
Synonyms | Hep-2, HEP-2, HEp-2/HeLa, Hep 2, Hep2, HEp2, HEP2, H.Ep.-2, H.Ep. #2, H.Ep. No. 2, Hep II, Human Epidermoid carcinoma #2, Human Epithelioma-2 |
Details of the HEp-2 cell line
Age | 30 years |
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Gender | Female |
Ethnicity | African American |
Morphology | Epithelial-like |
Growth properties | Monolayer, adherent |
Documentation
Citation | HEp-2 (Cytion catalog number 300397) |
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Biosafety level | 1 |
Genetic profile
Isoenzymes | G6PD, A |
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Reverse transcriptase | Negative |
Products | Keratin |
Handling of Hep2 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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Medium supplements | Supplement the medium with 10% FBS and 1% NEAA |
Passaging solution | Accutase |
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 1:10 is recommended |
Seeding density | 1 x 10^4 cells/cm^2 |
Fluid renewal | 2 to 3 times per week |
Freezing 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. |
Handling of cryopreserved cultures |
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Quality control
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: 9,10
D13S317: 12,13.3
D16S539: 9,10
D5S818: 11,12
D7S820: 8,12
TH01: 7
TPOX: 8,12
vWA: 16,18
D3S1358: 15,18
D21S11: 27,28
D18S51: 16
Penta E: 7,17
Penta D: 8,15
D8S1179: 12,13
FGA: 18,21
PEZ6: WT51
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Required products
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