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hTert-Human Renal Medullary Epithelial Cells


Item#:
cAP-0138hTert
Size Price Quantity
1.0 Frozen Vial $5,100.00
Name of Products hTert-Human Renal Medullary Epithelial Cells (HRMEPs)
Catalog NocAP-0138hTert
Product FormatFrozen Vial
Cell Number> 5x10[5] cells/vial
Human Renal Medullary Epithelial Cells (HRMEPs, cAP-0138) are initiated by digestion of minced human kidney medullary tissue from a healthy donor. HRMEPs are separated from a mixture of cell populations. hTert-HRMEPs were selected from HRMEPs by puromycin after being infected by hTert-expressing lentiviruses under the control of CMV promoter. Epithelial Growth Medium (EpGM) formulation-B (cAP-45C) containing FBS and growth supplement) is recommended for cell culture and these cells can be maintained in vitro long term for > 20 passages when cultured following the detailed protocol described below).
1. HRMEPs are tested negative for HIV-1, HBV, HCV, and mycoplasm.
2. Positive for Pan-Cytokeratin.
3. Negative for y-glutamyltransferase-1 (GGT-1).
The 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, 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 containing 10ml full media; spin down the cell @ 1000rpm for 10min, and resuspending the cells with 5ml of full medium, before transfering the cells in the pre-coated T25 flask. The cells usually become confluent within 2-3days.
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 1000rpm for 10 min.
G)Resuspend the cell pellet with 10 or 15ml Full medium and transfer 5 ml each into 2 or 3 pre-coated T25 flasks (for 1/2 to 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).
Quick Coating SolutioncAP-01
Epithelial Growth Medium (EpGM) formulation-B cAP-45C
ITS Solution (100x)cAP-26
(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.

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