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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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- Isoenzymes: G6PD, A
- Reverse transcriptase: Negative
- Products: Keratin
- cultureMedium: EMEM (MEM Eagle), w: 2 mM L-Glutamine, w: 2.2 g/L NaHCO3, w: EBSS (Cytion article number 820100a)
- supplements: Supplement the medium with 10% FBS and 1% NEAA
- dissociationReagent: 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.
- seedingDensity: 1 x 104 cells/cm2
- fluidRenewal: 2 to 3 times per week
- postThawRecovery: After thawing, plate the cells at 5 x 104 cells/cm2 and allow the cells to recover from the freezing process and to adhere for at least 24 hours.
- freezeMedium: 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.
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