1) Coating T25 flasks. Add 2 ml AlphaBioCoat (AC001) into a T25 flask and ensure entire interior surface is coated with the solution. After 30 minutes, dispose of AlphaBioCoat (AC001) by aspiration. Gently rinse and aspirate flask with Phosphate Buffer Solution (1XPBS-‐001). The flask is now ready for use (no need for overnight incubation when coated with AC001) 2) If you are using the coated flask the same day, add about 4 ml of Pericyte-‐Growth media to the coated flask. If the media changes color from pink to yellow, aspirate and discard the media. Add 4ml of fresh media to the coated flask. 3) Thaw the cells in a 37°C water bath. Once you see a small amount of ice left in the vial, spray the vial with 70% Ethanol and wipe it down. 4) Transfer the vial into your Biosafety cabinet. 5) Using a 2 or 5ml pipet, pipet the cells out of the vial. 6) Transfer your cell suspension in to your coated plate that have the 4 ml media in it. 7) You should have a total working volume of 5ml of cell suspension in the flask; close the cap. Make sure cells are evenly distributed in the flask by moving the flask left and right five times. Move it up and down for and additional five times. 8) Place flask in a 37°C incubator with 5% C02. If flask is not vented, please loosen cap. 9) Change media after 48 hours.
General Information
RFP expressing human brain microvascular pericytes were isolated from normal human brain tissue. Puromycin resistant, RFP-‐HBMVPCs are selected from RFP-‐Expressing Lentiviral after transfection with RFP-‐HBMVPCs lentiviral particle. Cells are shipped in proliferating culture or frozen vial with a confluence of > 90% (cells are provide @ passage 5). Pericyte-‐Growth medium (AGPM-‐01) containing 5% serum and growth supplement is recommended for culture. Cells have an average population doubling level >10 when cultured. When you receive the cells, leave the flask in 37°C C02 incubator for 1 hour. Then, replace the transport medium with fresh Pericytes-‐Growth medium (AGPM-‐01). Let the cells grow for 24 hours before subculture.
Tissue
Brain
Shipping
-80°C
Size
1ml frozen Vial
Product Format
Frozen vial
Product Use
Product Use RFP Expressing human brain microvascular Pericytes are for research use only Shipping Status Frozen vial
Storage
liquid Nitrogen Vapor Phase
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