| 產品名稱 |
HEK001 |
| 商品貨號 |
B164589 |
| Organism |
Homo sapiens, human |
| Tissue |
skin |
| Cell Type |
keratinocyte; human papillomavirus 16 (HPV-16) E6/E7 transformed |
| Product Format |
frozen |
| Morphology |
epithelial |
| Culture Properties |
adherent |
| Biosafety Level |
2 [Cells contain human papilloma viral 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. |
| Age |
65 year old |
| Gender |
male |
| Storage Conditions |
liquid nitrogen vapor phase |
| Derivation |
The HEK001(human epidermal keratinocyte 001) cell line was established in 1996. Cells from the scalp of a 65 year old male were transfected with plasmid p1321(a generous gift from Dr. Karl Munger, Harvard Medical School). Plasmid p1321 contains human papillomavirus (HPV) 16 E6 and E7 genes. |
| Clinical Data |
male
65 years |
| Genes Expressed |
keratin |
| Cellular Products |
keratin |
| Comments |
HEK001 cells express keratin 14 but not keratin 10 suggesting that they are proliferating basal-type keratinocytes. |
| Complete Growth Medium |
Keratinocyte-Serum Free medium (GIBCO-BRL 17005-042) with 5 ng/ml human recombinant EGF and 2mM L-glutamine (without bovine pituitary extract and without serum)
|
| 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 Ca++/Mg++ free Dulbecco's phosphate-buffered saline (D-PBS) or 0.05% (w/v) Trypsin - 0.53 mM EDTA solution to remove all traces of serum, which contains trypsin inhibitor.
- Add 2.0 to 3.0 mL of Trypsin-EDTA solution to the 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 medium with 10% FBS added and aspirate cells by gently pipetting.
- To remove trypsin-EDTA solution, transfer cell suspension to centrifuge tube and spin at approximately 125 x g for 5 to 10 minutes.
- Discard supernatant and resuspend cells in fresh growth medium. Add appropriate aliquots of cell suspension to new culture vessels.
- Incubate cultures at 37°C.
Subcultivation Ratio: A subcultivation ratio of 1:3 to 1:4 is recommended
Medium Renewal: Add fresh medium twice a week.
Note: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 13 in Culture Of Animal Cells: A Manual Of Basic Technique by R. Ian Freshney, 5th edition, published by Wiley-Liss, N.Y., 2005.
|
| Cryopreservation |
Freeze medium: Dulbecco's Modified Eagle's Medium, 70%? fetal bovine serum, 20%? DMSO, 10%. Storage temperature: liquid nitrogen vapor phase |
| Culture Conditions |
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 37°C |
| STR Profile |
Amelogenin: X,Y CSF1PO: 11 D13S317: 8 D16S539: 12,13 D5S818: 10,13 D7S820: 12,13 THO1: 6,9 TPOX: 8,9 vWA: 14 |
| Name of Depositor |
PB Sugerman, M Bigby |
| Deposited As |
human |
| Year of Origin |
1996 |
| References |
Sugerman PB, Bigby M. Preliminary functional analysis of human epidermal T cells. Arch. Dermatol. Res. 292: 9-15, 2000. PubMed: 10664009
|
