| 產(chǎn)品名稱 |
CCD 1103 KIDTr |
| 商品貨號(hào) |
B164143 |
| Organism |
Homo sapiens, human |
| Tissue |
kidney |
| Cell Type |
human papillomavirus 16 (HPV-16) E6/E7 transformed |
| Product Format |
frozen |
| Morphology |
epithelial |
| Culture Properties |
adherent |
| Biosafety Level |
2 [Cells contain Human Papilloma viral 16 (HPV16) 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. |
| Age |
115 days gestation |
| Derivation |
Fetal kidney was digested with collagenase: trypsin mixture. The digestion products were plated in REGM (Renal Epithelial Cell Growth Medium which includes 0.5% FBS) from Clonetics. Passage 3 cells were transformed with a retrovirus vector (LXSN16E6E7 produced by ATCC CRL-2203) in the presence of polybrene. The transformed cells are resistant to G418 and sensitive to Hygromycin B. After 55 population doublings, the cells continue dividing and retain cuboidal morphology. E6E7 sequences were detected by PCR in cells at passage 15. |
| Oncogene |
E6/E7 + |
| Genes Expressed |
keratin; acid phosphatase |
| Cellular Products |
keratin; acid phosphatase |
| Virus Susceptibility |
Human Coxsackievirus B 5
Human Coxsackievirus A 9
Measles virus
Human poliovirus 1
Human respiratory syncytial virus
|
| Comments |
This line stains positively with antibody for cytokeratin and is acid phosphatase positive.
After 55 population doublings, the cells continue dividing and retain cuboidal morphology. E6E7 sequences were detected by PCR in cells at passage 15.
Major Histocompatibility Complex class I or II molecules were not expressed on these cells, but PCR analyses revealed presence of the genes for directing synthesis of HLA antigens.
CCD 1103 KIDTr is sensitive to: Coxsackievirus A9 and B5, measles virus, poliovirus 1 and respiratory syncytial virus (RSV). |
| Complete Growth Medium |
The base medium for this cell line is ATCC-formulated F-12K Medium, Catalog No. 30-2004. To make the complete growth medium, add the following components to the base medium: fetal bovine serum to a final concentration of 10%.
|
| Subculturing |
Volumes used in this protocol are for 75 cm2 flask; 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 0.25% (w/v) Trypsin-0.53mM EDTA 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 supension to new culture vessels.
- Incubate cultures at 37°C.
Subcultivation Ratio: 1:3 to 1:5.
Medium Renewal: Two to three times weekly.
Note: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 10 in Culture of Animal Cells, a manual of Basic Technique by R. Ian Freshney, 3rd edition, published by Alan R. Liss, N.Y., 1994.
|
| Cryopreservation |
culture medium 95%; DMSO, 5% |
| Culture Conditions |
Temperature: 37°C |
| Name of Depositor |
A Thompson |
| References |
Zabrenetzky V, et al. The isolation, immortalization and characterization of human fetal keratinocytes. In Vitro Cell. Dev. Biol. 33: Part II, p. 34A, 1997.
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