Human Sox17 Expressing Embryonic Stem Cells | T2513
- 5 to 7 Days Shipment
Sox17 is a transcription factor that regulates endodermal differentiation of stem cells in the developing human embryo. The endoderm is one of the primary germ layers that appears early in embryonic developement. The endoderm gives rise to the lining of the digestive tract, respiratory tract and endocrine organs. Human Sox17 Expressing Embryonic Stem Cells are endoderm progenators and exhibit markers of the deffinitive endoderm. Sox17 plays a critical role in general organogenesis of the endoderm during embryonic developement and this cell line remains able to undergo hepatic and pancreatic differentiation.
The base medium for this cell line is Prigrow III medium available at abm, Cat. No. TM003. To make the complete growth medium, add the following components to the base medium: KnockOut™ Serum Replacement (Gibco) to a final concentration of 15% and Penicillin/Streptomycin Solution to a final concentration of 1%.
Change media every 2-3 days.
Carbon dioxide (CO2): 5%, Temperature: 37.0°C.
GATA4, GATA6, HNF4A, FOXA2, CXCR4, CER, GSC, DLX5, ECAD and FOXQ1
1. Pre-warm the Trypsin-EDTA (TM050) to room temperature.
2. Carefully aspirate the culture media from the culture vessel without disturbing the cell monolayer.
3. Add pre-warmed Trypsin-EDTA to the culture vessel. Gently rock the culture vessel to ensure complete coverage of the Trypsin-EDTA over the cells.
4. Observe the cells under a microscope to confirm they are dissociating from each other and are rounding up. Gently tap the culture vessel from several sides to promote cell detachment. Cells that are difficult to detach can be put in 37°C for several minutes to facilitate dispersal.
5. When majority of the cells have detached, add an equal volume of Trypsin Neutralizing Solution into the culture vessel to neutralize the trypsin-EDTA. Gently swirl or pipette the culture suspension to ensure the neutralization is complete.
6. Transfer the culture suspension to a sterile centrifuge tube.
7. Centrifuge the cell suspension at 1500 rpm for 3 minutes. The actual centrifuge duration and speed may vary depending on the cell type.
8. Aspirate the supernatant after checking all cells are pulled down into the pellet. Re-suspend the cell pellet in pre-warmed fresh complete media.
9. Pre-warm new culture vessels to 37°C*. Seed cells at the recommended seeding density.
10. Place the newly seeded culture vessel in a 37°C*, 5% CO2 incubator. Incubate for at least 24 – 48 hours before processing the cells for downstream experiments.
11. Renew the culture media every 2-3 days if the cells have not reached 80% confluency.
* Unless a different temperature is recommended on the data sheet.
BioSafety Level II