GFP Stably Expressing Immortalized Human Astrocytes, Fetal - hTERT Cell Line | T3962
- 5 to 7 Days Shipment
abm | GFP Stably Expressing Immortalized Human Astrocytes, Fetal - hTERT Cell Line | T3962
Immortalized human astrocytes stably expressing GFP
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 15mL sterile conical tube containing 5mL 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.
The base medium for this cell line is Prigrow IV medium available at abm
Drug selection by selection marker
10⁶ cells/1.0 ml