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| Name of Products | ACE2-GFP Expressing Human Pulmonary Lobar Bronchial Epithelial Cells (HPuLBEPCs-ACE2-GFP) |
| Catalogue Number | cAP-0123 |
| Product Format | Frozen Vial |
| Cell Number | >5x10[5] cells/vial |
| Huma Pulmonary Lobar Bronchial Epithelial Cells (HPuLBEPCs, cAP-0123) cells were isolated from normal human lung Lobar Bronchial tissue. ACE2-GFP Expressing Human Pulmonary Lobar Bronchial Epithelial Cells were selected from puromycin resistant HPuLBEPCs after infected with lentivirses expressing ACE2-GFP. The cells are shipped in frozen vials (the cells are provided @ passage 3). Human Airway Epithelial Growth Medium (HAEPGM, composed of Airway Epithelial Cell Basal Medium (ATCC Cat NO PCS-300-030) and Bronchial Epithelial Cell Growth Kit (ATCC CAT NO PCS-300-040) is recommended for cell culture and these cells have a minimum average population doubling levels > 8 when cultured following the detailed protocol described below). |
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| 1. | Pan-Cytokeratin Positive |
| 2. | E-7 negative |
| 3. | Typical cobblestone monolayer morphology in culture |
| 4. | HPuLBEPCs are negative for HIV-1, HBV, HCV, and mycoplasma |
| Cells are offered for Research Use Only. |
| Frozen Vials in 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 T25 flasks- Add 2-3ml each Quick Coating Solution (cAP-01) into a T25 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 a 15ml falcon tube with 10ml of Universal Growth Medium (cAP-01B) and spin @1000rpm for 10 mins. Dispose the supernatant and resuspend the cells in 5 ml of HAEPGM and transfer the cells into the pre-coated T25 flask, the cells usually become 90% confluent within 3- 5 days and ready to be passaged. |
| C) | To passage the cells, rinse the cells in a T25 flask with 5ml HBSS (RT) twice; then add 2ml Trypsin/EDTA (RT) (cAP-23) into one T25 flask; gently dispose the excessive Trypsin/EDTA solution within 20 seconds by aspiration. |
| D) | Leave the T25 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 with 800g centrifugation for 5 mins. |
| G) | Re-suspend the cell pellet with 15ml HAEPGM and transfer 5 ml each into 3 pre-coated T25 flasks (for 1/3 subculture ratio). |
| H) | Change medium every 2 or 3days and the cells usually become confluent within 7 days (when split at a 1/3 ratio). |
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| Quick Coating Solution | cAP-01 |
| Universal Growth Medium | cAP-01B |
| 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 |
| Airway Epithelial Cell Basal Medium | ATCC Cat NO PCS-300-030 |
| Bronchial Epithelial Cell Growth Kit | ATCC CAT NO PCS-300-040 |
| 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. |
