Catalog Number RFP1Product Name RFP Expressing Human retinal microvascular endothelialcells (RFP-HRMECs)Storage Liquid NitrogenProduct Format Frozen vialCells Number >90% confluent in Frozen vial*Caution: The handling of human derived products has the potential to be biologically hazardous. All Cell strains tested negative for HIV, HBV,and HCV DNA in diagnostic tests. Proper precautions must be taken to avoid exposure. Always wear proper protective equipment (Gloves,safety glasses, etc.) when handling these materials. We recommend following the universal procedures for handling products of human originas the minimum precaution against contamination.GENERAL INFORMATIONRFP expressing human retinal microvascular endothelial cells were isolated from normal human retinaltissue and transfected with RFP- lentiviral particles at passage 1. Zeocin resistant RFP- HRMECs isshipped in proliferating culture or frozen vial with a confluence of > 90%( cells are provide @ passage 4-5). ENDO-Growth medium (EGM-2102) containing 5% serum and growth supplement is recommendedfor culture. Cells have an average population doubling level >16 when cultured. When you receive thecells, leave the flask in 37°C C02 incubator for 1 hour. Then, replace the transport medium with freshENDO-Growth medium (EGM-2102). Let the cells grow for 24 hours before subculture.CELL CHARACTERIZATIONCytoplasmic VWF/ factor VIII >95% positive by immunofluorescenceCytoplasmic uptake of Di-I-Ac-LDL >95% positive by immunofluorescenceCytoplasmic PECAM1 >95% positive by immunofluorescenceRFP expressing human retinal microvascularendothelial cells are negative forHIV-1, HBV, HCV, and mycoplasmaPRODUCT USE AND SHIPPING STATUSProduct Use RFP expressing human retinal microvascularendothelial are for research use onlyShipping Status Frozen vial
Frozen1) Coating T25 flasks. Add 2 ml AlphaBioCoat (AC001) into a T25 flask and ensure entire interiorsurface is coated with the solution. After 30 minutes, dispose of AlphaBioCoat (AC001) byaspiration. Gently rinse and aspirate flask with Phosphate Buffer Solution (1XPBS-001). The flaskis 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 Endo-Growth media to thecoated 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 thevial 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 it7) 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.