Immortalized human astrocytes stably expressing GFP
Quantity: 1 million cells/1.0 ml
Organism: Human
Source Organ: Brain
BioSafety Level: II
Growth Properties: Adherent
Morphology: Multipolar
Markers: Neomycin
Propagation: The base medium for this cell line is Prigrow IV medium available at abm, Cat. No. TM004. To make the complete growth medium, add the following components to the base medium: fetal bovine serum (TM999)* to a final concentration of 10%, 10 ng/ml Human EGF (Z100135), 1% L-glutamine (G275), and Penicillin/Streptomycin Solution (G255) to a final concentration of 1%. Carbon dioxide (CO2): 5%, Temperature: +37.0°C.* abm does not recommend to use heat-inactivated FBS for cell culture unless specified otherwise.
Freeze Thaw: 1. Pre-warm complete media in a +37°C waterbath.2. Remove the cryopreserved vial from the liquid nitrogen storage tank.3. Thaw the cells quickly by placing the lower half of the vial into the +37°C water bath and remove after 60 seconds. There should still be a few ice crystals left after thawing. It is important not to over-thaw the cryovials as the presence of DMSO is toxic to the cells.4. Re-suspend the cells in the vial and transfer the cell suspension into a 15 ml sterile conical tube containing 5 ml complete media.5. Centrifuge cells at 200x g for 3 minutes to pellet.6. Aspirate out the media, leaving cell pellet undisturbed.7. Re-suspend the cell pellet in fresh, pre-warmed culture media and perform a viable cell count.8. Transfer the cells into a culture vessel using the recommended seeding density. Gently rock the culture vessel to distribute the cells evenly. Table 1 provides general guidelines to the volume of culture media needed for a range of culture vessels.Note:The size of the culture vessel is subjected to the seeding density of the cell line (available on the corresponding cell line webpage “seeding density” section). Otherwise, we recommend thawing entire cryopreserved content into T25 flask with instructed medium condition from the propagation section of the product page.9. Incubate the culture at +37°C, 5% CO2 or another recommended culture environment for the specific cell line. Incubate for at least 24 hours before processing the cells for downstream experiments.Note: Where applicable, we recommend selection drug addition after cells recovered from thawing. Directly adding selection drug while thawing could lead to stressed cells and lower viability.
Quality Control: Drug selection by selection marker
Shipping Conditions: Dry Ice
Storage Conditions: -180°C