| 產品名稱 |
2A3 |
| 商品貨號 |
B185404 |
| Organism |
Homo sapiens, human |
| Tissue |
pharynx |
| Product Format |
frozen |
| Morphology |
epithelial-like |
| Culture Properties |
adherent |
| Biosafety Level |
2 [Cells contain human papillomavirus (HPV) DNA sequences]
Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country. |
| Disease |
squamous cell carcinoma |
| Age |
56 years old |
| Gender |
male |
| Ethnicity |
Caucasian |
| Storage Conditions |
liquid nitrogen vapor phase |
| Karyotype |
(P16) hypodiploid to hypertriploid with modal number = 64 |
| Derivation |
The 2A3 cell line was derived by transfecting ATCC HTB-43 (FaDu) with E6 and E7 genes using PA317 LXSN 16E6/E7. The pLXSN16E6E7 vector contained the human papilloma virus (HPV) type 16 E6 and E7 genes under control of the Moloney murine leukemia virus (MoMuLV) promoter-enhancer sequences. The vector also contained a gene encoding resistance to neomycin transcribed from the SV40 promoter. |
| Clinical Data |
56 years
Caucasian
male
|
| Tumorigenic |
Yes |
| Effects |
Yes, in nude mice; forms well differentiated epidermoid carcinoma (grade I) |
| Comments |
The 2A3 cell line was derived by transfecting ATCC HTB-43 (FaDu) with E6 and E7 genes using PA317 LXSN 16E6/E7. The pLXSN16E6E7 vector contained the human papilloma virus (HPV) type 16 E6 and E7 genes under control of the Moloney murine leukemia virus (MoMuLV) promoter-enhancer sequences. The vector also contained a gene encoding resistance to neomycin transcribed from the SV40 promoter. This stably transfected head and neck cell line may be used as a basis for animal models for drug development as it was 100% tumorigenic in nude mice while continuing to express E6 oncoprotein in vivo.
|
| Complete Growth Medium |
The base medium for this cell line is ATCC-formulated Dulbecco's Modified Eagle's Medium, Catalog No. 30-2002.
To make the complete growth medium, add the following components to the base medium:
0.2 mg/ml G -418
fetal bovine serum to a final concentration of 10%
|
| Subculturing |
Volumes used in this protocol are for 75 cm2 flasks; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.
- Remove and discard culture medium. Briefly rinse the cell layer with Ca++/Mg++ free Dulbecco's phosphate-buffered saline (D-PBS) (ATCC 30-2200) or 0.25% Trypsin – 0.53mM EDTA (ATCC 30-2101) solution to remove all traces of serum which contains trypsin inhibitor.
- 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.
- Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.
- Add appropriate aliquots of the cell suspension to new culture vessels. Incubate cultures at 37°C.
Subcultivation Ratio: 1:3 to 1:6 is recommended
Medium Renewal: 2 to 3 times per week |
| Cryopreservation |
Culture medium, 95%; DMSO, 5% |
| Culture Conditions |
Temperature: 37°C |
| STR Profile |
Amelogenin: None detected
CSF1PO: 12
D13S317: 8, 9
D16S539: 11
D5S818: 12
D7S820: 11, 12
THO1: 8
TPOX: 11
vWA: 15, 17 |
| Isoenzymes |
AK-1, 1
ES-D, 1
G6PD, B
GLO-I, 2
Me-2, 2
PGM1, 2
PGM3, 1 |
| Population Doubling Time |
51 hours |
| Name of Depositor |
E Dadachova, Ph.D. |
| Year of Origin |
June 2010 |
| References |
Harris M, et al. Radioimmunotherapy of experimental head and neck squamous cell carcinoma (HNSCC) with E6-specific antibody using a novel HPV-16 positive HNSCC cell line. Head Neck Oncol. 3(1): 9, 2011. PubMed: 21314983
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