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ACE2-GFP Expressing Human Alveolar Epithelial Cells


Item#:
cAP-0208ACE2-GFP
Size Price Quantity
1.0 Frozen Vial $850.00
Name of Products ACE2-GFP Expressing Expressing Human Alveolar Epithelial Cells
Catalogue NumbercAP-0208ACE2
Product FormatFrozen Vial
Cell Number> 5x10[5] cells/vial
Human Alveolar Epithelial Cells (HAlvEPCs, cAP-0208) are isolated from normal human adult lung tissue. ACE2-GFP Expressing HAlvEPCs (under the control of CMV promoter) were selected from GFP-fused (at C-terminal) ACE2 expressing lentiviruses infected HAlvPCs by 4ug/ml puromycin and the stable cells are shipped in frozen vials (the cells are provided @ passage 3). Human Alveolar Epithelial Cells Growth Medium (HAPCGM, from ATCC, Epithelial Cell Basal Medium (PCS-300-030) plus Epithelial Cell Growth Kit (PCS-300-040)) is recommended for cell culture and these cells have a minimum average population doubling levels > 15 when cultured following the detailed protocol described below). HAlvEPCs 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.
A)Pre-coating of T25 flasks- Add 2ml each Quick Coating Solution (cAP-01) into a T25 flask to cover the whole surface of the flask, 5 mins later, disposing excessive coating solution by aspiration and the flask is ready to be used (although solution containing other extracellular matrices, 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 15ml Falcon tube with 10ml of HAPCGM, Spin down the cells with 800g for 8 mins, resuspend the cells in 5ml of HAPCGM and transfer the cells into a pre-coated T25 flask, and culture the cells in a 37C and 5%CO2 incubator. The cells usually become confluent after 2-3 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 8 mins.
G)Re-suspend the cell pellet with 15ml HAPCGM medium and transfer 5 ml each into 3 pre-coated T25 flasks (for 1 to 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).
I)To prepare quiescent cells, when cells are nearly confluent. Related Products#
Quick Coating SolutioncAP-01
HBSS w/o Ca2+, Mg2+cAP-11
Cell Freezing Solution (FBS)cAP-22
Cell Freezing Solution (Non-FBS)cAP-22B
Trypsin/EDTA SolutioncAP-23
Trypsin Neutralization SolutioncAP-28
ITS (100x)cAP-26
L-Glutamine-MAXIMUM (100x) cAP-27
Human Plasma Fibronectin SolutioncAP-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.
SPECIAL NOTES: 1. We strongly advise our customers using our own medium and related accessory products; 2. The quality of cells, in term of their growth features stated in our datasheets, will not be waranteed if NON-ANGIO-PROTEOMIE's Cell Growth Medium is used; 3.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.
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