RFP Expressing Human Brain Astrocytes

Catalog Number HBMP203
Product Name RFP Expressing Human Brain Astrocytes (RFP-HBA)
Storage 37°C C0₂ incubator or Liquid Nitrogen
Product Format Proliferating culture or Frozen vial
Cells 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 of
human origin as the minimum precaution against contamination.
GENERAL INFORMATION
RFP Expressing Human Brain Astrocytes were isolated from normal human brain
Astrocytes and transfected with GFP-Lentiviral particles. Puromycin resistant, RFP-HBA is
shipped in proliferating culture or frozen vial with a confluence of > 90% (cells are
provided at passage 3). Astrocyte-Growth medium (CAT#AGPM-03) containing 5% fetal
bovine serum and growth supplement is recommended for culture. Cells have an average
population doubling level >8 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 AstrocyteGrowth medium (CAT#AGPM-03). Let the cells grow for 24 hours before subculture.
CELL CHARACTERIZATION
Cytoplasmic GFAP >98% positive by immunofluorescence
RFP Expressing Human Brain Astrocytes are
negative for
HIV-1, HBV,HCV, and mycoplasma
PRODUCT USE AND SHIPPING STATUS
Product Use RFP Expressing Human Brain Astrocytes
cells are for research use only
Shipping Status Proliferating cells in T25 Flask or Frozen
vial

SUBCULTURE PROTOCOL
*If Cells arrive frozen:
When you receive the cells in a frozen vial, you can transfer the vial of cells into -80°C
freezer 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
ensure entire interior surface is coated with the solution. After 30 minutes, dispose
of AlphaBioCoat (CAT# 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 CAT# AC001)
2) If you are using the coated flask the same day, add about 4 ml of Astrocyte-Growth
media (CAT# AGPM-03) 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.