HEp-2 cell line    【中文名稱】：    人喉表皮樣癌細胞    

The HEp-2 cell line contains HeLa marker chromosomes and were derived via HeLa contamination. This line was originally thought to be derived from an epidermoid carcinoma of the larynx but was subsequently found – based on isoenzyme analysis, HeLa marker chromosomes, and DNA fingerprinting – to have been established via HeLa cell contamination. The cells are positive for keratin by immunoperoxidase staining. BioVector confirmed this cell line is positive for the presence of human papilloma viral DNA sequences via PCR.

Product category

Human cells

Organism

Homo sapiens, human

Morphology

epithelial

Disease

Carcinoma

Applications

3D cell culture

Characteristics

Growth properties

Adherent

Derivation

Cells of this line contain HeLa marker chromosomes, and were derived via HeLa contamination. This line was originally thought to be derived from an epidermoid carcinoma of the larynx, but was subsequently found, based on isoenzyme analysis, HeLa marker chromosomes, and DNA fingerprinting, to have been established via HeLa cell contamination. The cells are positive for keratin by immunoperoxidase staining.

Karyotype

Occasional polyploids. Several marker chromosomes were observed along with frequent minutes, and often 2 large chromosomes with subterminal centromeres.HeLa Marker Chromosomes: One copy of M2, two-four copies of M3 and one copy of M4 as revealed by G-banding patterns.

Virus susceptibility

Human adenovirus 3

Human poliovirus 1

Vesicular stomatitis virus

Genes expressed

keratin

Isoenzymes

G6PD, A

Comments

Cells of this line contain HeLa marker chromosomes, and were derived via HeLa contamination. This line was originally thought to be derived from an epidermoid carcinoma of the larynx, but was subsequently found, based on isoenzyme analysis, HeLa marker chromosomes, and DNA fingerprinting, to have been established via HeLa cell contamination. The cells are positive for keratin by immunoperoxidase staining. BioVector confirmed this cell line is positive for the presence of human papilloma viral DNA sequences via PCR.

Handling information

Unpacking and storage instructions

1. Check all containers for leakage or breakage.

2. Remove the frozen cells from the dry ice packaging and immediately place the cells at a temperature below -130°C, preferably in liquid nitrogen vapor, until ready for use.

Complete medium

The base medium for this cell line is BioVector-formulated Eagle's Minimum Essential Medium, Catalog No. 30-2003. To make the complete growth medium, add the following components to the base medium: fetal bovine serum to a final concentration of 10%.

Temperature

37°C

Atmosphere

95% Air, 5% CO2

Handling procedure

To insure the highest level of viability, thaw the vial and initiate the culture as soon as possible upon receipt. If upon arrival, continued storage of the frozen culture is necessary, it should be stored in liquid nitrogen vapor phase and not at -70°C.  Storage at -70°C will result in loss of viability.

1. Thaw the vial by gentle agitation in a 37°C water bath. To reduce the possibility of contamination, keep the O-ring and cap out of the water.  Thawing should be rapid (approximately 2 minutes).

2. Remove the vial from the water bath as soon as the contents are thawed, and decontaminate by dipping in or spraying with 70% ethanol. All of the operations from this point on should be carried out under strict aseptic conditions.

3. It is recommended that the cryoprotective agent be removed immediately.  Centrifuge the cell suspension at approximately 125 x g for 5 to 10 minutes.  Discard the supernatant and resuspend the cell pellet in an appropriate amount of fresh growth medium.

4. Transfer the vial contents to an appropriate size vessel.  It is important to avoid excessive alkalinity of the medium during recovery of the cells.  It is suggested that, prior to the addition of the vial contents, the culture vessel containing the growth medium be placed into the incubator for at least 15 minutes to allow the medium to reach its normal pH (7.0 to 7.6).

5. Incubate the culture at 37°C in a suitable incubator.  A 5% CO2 in air atmosphere is recommended if using the medium described on this product sheet.

Subculturing procedure

Volumes used in this protocol are for 75 cm2 flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.

1. Remove and discard culture medium

2. Briefly rinse the cell layer with 0.25% (w/v) Trypsin-0.53mM  EDTA solution to remove all traces of serum which  contains trypsin inhibitor.

3. Add 2.0 to 3.0 mL of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 15 minutes). Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach.  Cells that are difficult to detach may be placed at 37°C to facilitate dispersal.

4. Add 6.0 to 8.0 mL of complete growth medium and aspirate  cells by gently pipetting.

5. Add appropriate aliquots of the cell suspension to new culture vessels.

6. Incubate cultures at 37°C.

Subcultivation Ratio: A subcultivation ratio of 1:4 to 1:10 is recommended

Medium Renewal: 2 to 3 times per week

Quality control specifications

Mycoplasma contamination

Not detected

Virus testing

Human papillomavirus (HPV): Detected

STR profiling

Amelogenin: X

CSF1PO: 9,10

D13S317: 12,13.3

D16S539: 9,10

D5S818: 11,12

D7S820: 8,12

THO1: 7

TPOX: 8,12

vWA: 16,18

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