| 產品名稱 |
CCD 1102 KERTr |
| 商品貨號 |
B164142 |
| Organism |
Homo sapiens, human |
| Tissue |
skin |
| Cell Type |
keratinocyte; human papillomavirus 16 (HPV-16) E6/E7 transforme |
| Product Format |
frozen |
| Morphology |
epithelial |
| Culture Properties |
adherent |
| Biosafety Level |
2 Cells may contain the human papilloma viral (HPV) 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 |
112 days gestation fetus |
| Applications |
6E7 sequences were detected by PCR in cells at passage 18.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. 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. |
| Storage Conditions |
liquid nitrogen vapor phase |
| Karyotype |
hyperdiploid; about 55% of cells contain 45 to 50+ chromosomes |
| Derivation |
Rockville Maryland, United States |
| Antigen Expression |
epithelial specific antigen; Homo sapiens, expressed (Epithelial specific antigen was detected using antibody produced by the Ep16 hybridoma (ATCC HB-155).) (Epithelial specific antigen was detected using antibody produced by the Ep16 hybridoma (ATCC HB-155).) |
| Oncogene |
E6/E7 + |
| Genes Expressed |
E6/E7 +,epithelial specific antigen; Homo sapiens, expressed (Epithelial specific antigen was detected using antibody produced by the Ep16 hybridoma (ATCC HB-155).) |
| Comments |
After 50 population doublings, the cells continue dividing and retain cuboidal morphology.
E6E7 sequences were detected by PCR in cells at passage 18.
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.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.
|
| Complete Growth Medium |
These cells are grown in Keratinocyte-Serum Free Medium (Gibco 17005-042) with added Keratinocytes Supplements (Gibco 37000-015) including Bovine Pituitary Extract (BPE; Gibco 13028-014) and human recombinant epidermal growth factor (EGF; Gibco 10450-013) further supplemented with:
- Additional 30 ng/ml human recombinant epidermal growth factor (EGF; BD cat# 354052)
Do not filter the complete medium.
This medium is formulated for use with a 5% CO2 in air atmosphere.
|
| 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 suspension to new culture vessels.
- Incubate cultures at 37°C.
Subcultivation Ratio: 1:3 to 1:6
Medium Renewal: Every 2 to 3 days
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 |
Ham?s F12 medium, 85%; fetal bovine serum,10% ; 5% DMSO. Cell culture tested DMSO is available as ATCC Catalog No. 4-X. |
| Culture Conditions |
Temperature: 37°C
Atmosphere: Air, 95%; Carbon dioxide (CO2), 5% |
| Isoenzymes |
G6PD, A-B (This lines has a rare human G6PD phenotype (A-B).) |
| Name of Depositor |
L Vilner, A Thompson |
| Passage History |
After 50 population doublings, the cells continue dividing and retain cuboidal morphology.
E6E7 sequences were detected by PCR in cells at passage 18.
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.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.
|
| Year of Origin |
November, 1995 |
| 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.
Fetal skin was digested with a collagenase-trypsin mixture. The digestion products were plated on collagen- fibronectin-BSA coated flasks in Keratinocyte Serum-Free Medium with epidermal growth factor and bovine pituitary extract. Passage 3 cells were transformed with a retrovirus vector (LXSN16E6E7 produced by ATCC CRL-2203) in the presence of polybrene.
This lines has a rare human G6PD phenotype (A-B).
Epithelial specific antigen was detected using antibody produced by the Ep16 hybridoma (ATCC HB-155).
Hay, R. J., Caputo, J. L., and Macy, M. L., Eds. (1992), ATCC Quality Control Methods for Cell Lines. 2nd edition, Published by ATCC.
Caputo, J. L., Biosafety procedures in cell culture. J. Tissue Culture Methods 11:223-227, 1988.
Fleming, D.O., Richardson, J. H., Tulis, J.J. and Vesley, D., (1995) Laboratory Safety: Principles and Practice. Second edition, ASM press, Washington, DC.
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