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| Name of Products | hTert-Human Epidermal Keratinocytes |
| Catalog Number | cAP-0212-hTert-CDK4 |
| Product Format | Frozen Vial |
| Cell Number | > 5x10[5] cells/vial |
| Human Epidermal Keratinocytes (cAP-0212) are isolated from normal human adult epidermal tissue. hTert-CDK4-Human Epidermal Keratinocytes were selected with 2ug/ml puromycin and Hygromycin (100ug/ml) after being infected sequentially with lentiviral particles expressing hTert or CDK4 under the control of CMV and the hTert-CDK4-Human Epiermal Keratinocytes are shipped in frozen vials (the cells are provided @ passage 3). Human Keratinocyte Growth Medium (KGM, cAP-67) is recommended for cell culture and these cells have a minimum average population doubling levels >40 when cultured following the detailed protocol described below). Human Epidermal Keratinocytes are negative for HIV-1, HBV, HCV, and mycoplasma |
| Cells are offered for Research Use Only. |
| Frozen Vials in a Dry Ice Package. |
| When you receive the dry ice package with cells in frozen vials, transfer the frozen vials of cells into a -80C freezer for short period storage or a liquid nitrogen tank for long-term storage. |
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| A) | Pre-coating of T75 flasks- Add 4ml each Quick Coating Solution (cAP-01) into a T75 flask to cover the whole surface of the flask, 5 mins later, dispose the excessive coating solution by aspiration and the flask is ready to be used (although solution containing other extracellular matrix, i.e. gelatin, collagen, and fibronectin, can be used, make sure to optimize the conditions in advance). |
| B) | Thaw the frozen cell vial in a 37C water bath first, and then transfer the cells into 10ml full medium in a 15 ml Falcon tube and spin down the cells @ 1000RPM for 8mins and transfer the cell pallet into the pre-coated T75 flask with 15ml of cAP-67 medium. Change the medium every two days until the cells become 85% confluent. |
| C) | To passage the cells (do not allow the cells to >90% confluent before passage), rinse the cells in a T25 flask with 10ml HBSS (RT) twice; then add 2ml Trypsin/EDTA (RT) (cAP-23) into one T75 flask; gently dispose the excessive Trypsin/EDTA solution within 20 seconds by aspiration. |
| D) | Leave the T75 flask with the cells at RT or 37C for 1 min (most cells usually will detach from the surface within 1-2 mins; or monitor the cells under a microscope until most of cells become rounded up, and then gently tap the flask against the bench surface, and the cells will move on the surface of the flask when monitoring under microscope. |
| E) | Add 5ml Trypsin Neutralization Buffer and spin down the cells @ 1000RPM centrifugation for 8 mins. |
| G) | Re-suspend the cell pellet with 30 or 45ml cAP-67 medium and transfer 15 ml each into 2 or 3 pre-coated T75 flasks (for 1/2 to 1/3 subculture ratio). |
| H) | Change medium every 2 or 3days and the cells usually become 85% confluent within 7 days (when split at a 1/3 ratio). |
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| Quick Coating Solution | cAP-01 |
| Keratinocyte Growth Medium | cAP-67 |
| HBSS w/o Ca2+, Mg2+ | cAP-11 |
| Cell Freezing Solution (FBS) | cAP-22 |
| Cell Freezing Solution (Non-FBS) | cAP-22B |
| Trypsin/EDTA Solution | cAP-23 |
| Trypsin Neutralization Solution | cAP-28 |
| ITS (100x) | cAP-26 |
| L-Glutamine-MAXIMUM (100x) | cAP-27 |
| Human Plasma Fibronectin Solution | cAP-42 |
| Handling human tissue-derived products is potentially bio-hazardous, despite tested negative for HIV, HBV and HCV DNA; nonetheless, proper precautions must be taken to avoid inadvertent exposure. |
For customers using our primary cell products, we strongly advise you using our own medium, although similar cell growth medium can be purchased and used for our primary cell products. |
The quality of cells, in term of their growth features stated in our datasheets, will not be guaranteed if NON-ANGIO-PROTEOMIE's Cell Growth Medium is used. |
Due to the sensitive nature of primary cells and cell lines, all quality related issues about the cells products must be reported back to us within ONE month period after receiving the products, no quality warranty (i.e. replacement of cells) will be provided after the ONE-Month period. Thank you for your understanding. |
