hTert-GFP-Mouse Glomerular Podocytes (hTert-GFP-mGloPods)

5x10[5] cells/vial

General Information# Mouse Glomerular Podocytes (mGloPods) were isolated from kidneys of 4-week old C57/Black6 mice and hTert-GFP-mGloPods were selected from mGloPods infected with hTert and GFP expressing lentiviruses with puromycin. A clone of hTert-GFP-mGloPods which maintain the foot process formation were isolated and supplied in frozen vials. Podocyte Growth Medium (cAP-44) is recommended for the expansion of hTert-GFP-mGloPods and the maintaining of foot process formation in vitro. And we warranty a minimum of 30 passages of the cells if cultured following the detailed protocol described below).
Characterization of the Cells#
1.:Positive for CD2AP and Nephrin
2.:Negative for CD31
3.: hTert-GFP-mGloPods are tested negative for common animal pathogen and mycoplasma.
Product Usage# The cells are offered for Research Use Only.
Handling of Arriving Cells# 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.
Protocols for Thawing the cells and subculture#
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.
B):Thaw the frozen cell vial in a 37C water bath first, and then transfer the cells into the pre-coated T25 flask with 10ml of cAP-44 medium, cells usually become confluent overnight 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 10 or 15ml cAP-44 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).
I):To prepare quiescent cells, when cells are nearly confluent, replace cAP-44 with Podocytes Basal Medium (PodBM, cAP-44B) containing 0.5%FBS for about 8-12hrs before your experiments.
Note# The purchase of this product conveys to the buyer the nontransferable right to use the purchased amount of the product and all replicates and derivatives for research purposes conducted by the buyer in his laboratory only.