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Catalog Number HBMP201Product Name GFP Expressing Human Brain Astrocytes (GFP-HBA)Storage 37°C C0₂ incubator or Liquid NitrogenProduct Format Proliferating culture or Frozen vialCells Number >90% confluent in T25 flask />5×10⁵ 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 ofhuman origin as the minimum precaution against contamination.GENERAL INFORMATIONGFP Expressing Human Brain Astrocytes were isolated from normal human brain Astrocytesand transfected with GFP-Lentiviral particles. Puromycin resistant, GFP-HBA is shipped inproliferating culture or frozen vial with a confluence of > 90% (cells are provide @ passage 3).Astrocyte-Growth medium (CAT# AGPM-03) containing 5% fetal bovine serum and growthsupplement is recommended for culture. Cells have an average population doubling level >8when cultured. When you receive the cells, leave the flask in 37°C C02 incubator for 1 hour.Then, replace the transport medium with fresh Astrocyte-Growth medium (CAT#AGPM-03).Let the cells grow for 24 hours before subculture.CELL CHARACTERIZATIONCytoplasmic GFAP >98% positive by immunofluorescenceGFP Expressing Human Brain Astrocytes arenegative forHIV-1, HBV,HCV, and mycoplasmaPRODUCT USE AND SHIPPING STATUSProduct Use GFP Expressing Human Brain Astrocytescells are for research use onlyShipping Status Proliferating cells in T25 Flask or Frozenvial
SUBCULTURE PROTOCOL*If Cells arrive frozen:When you receive the cells in a frozen vial, you can transfer the vial of cells into -80°Cfreezer for short period storage or a liquid nitrogen tank for long term storage.1) Coating T25 flasks. Add 2 ml AlphaBioCoat (CAT# AC001) into a T25 flask and ensureentire interior surface is coated with the solution. After 30 minutes, dispose ofAlphaBioCoat (CAT# AC001) by aspiration. Gently rinse and aspirate flask withPhosphate Buffer Solution (1XPBS-001). The flask is now ready for use (no need forovernight incubation when coated with CAT# AC001)2) If you are using the coated flask the same day, add about 4 ml of Astrocyte-Growthmedia (CAT# AGPM-03) to the coated flask. *If the media changes color from pink toyellow, 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 thecap. Make sure cells are evenly distributed in the flask by moving the flask left and rightfive 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.