KC-0352

293T-CCR6 Cell Line

Home » 293T-CCR6 Cell Line

Background

Chemokine receptor 6 (CCR6), also named CD196, is a CC chemokine receptor protein, mainly expressed on immature dendritic cells and memory T cells, and play an important role in B-linage cell maturation and different ion, and regulation in migration and recruitment of dendritic and T cells during inflammatory and immunological responses.

Specifications

Catalog Number:
KC-0352
Cell Line Name:
293T-CCR6 Cell Line
Host Cell Line:
Human HEK293T cell line
Description:
HEK293T cell line stable expressing exogenous human CCR6 gene
Quantity:
One vial of frozen cells (5×106 per vial)
Stability:
Stable in culture over a minimum of 10 passages
Application:
Drug screening and biological assays
Freeze Medium:
70% DMEM+20% FBS+10% DMSO
Propagation Medium:
DMEM+10%FBS+1µg/mL Puromycin
Selection Marker:
Puromycin
Morphology:
Epithelial
Subculture:
Split saturated culture 1:3-1:6 every 3 days; seed out at about 1-3 × 105 cells/mL
Incubation:
37 °C with 5% CO2
Storage:
Liquid nitrogen immediately upon receiving
Doubling Time:
Approximately 30 hours
Mycoplasma Status:
Negative

Cell Line Generation

293T human CCR6 cell line was generated using lentiviral vector expressing human CCR6 sequence.

Characterization

Figure: Characterization of CCR6 overexpressing in 293T stable clones using FACS

Application

Cell Resuscitation

1. Prewarm culture medium (DMEM + 10% FBS + 1μg/mL puromycin)in a 37°C water bath.
2. Thaw the frozen vial in a 37°C water bath for 1-2 minutes.
3. Transfer the vial into biosafety cabinet, and wipe the surface with 70% ethanol.
4. Unscrew the top of the vial and transfer the cell suspension gently into a sterile centrifuge tube containing 9.0mL complete culture medium.
5. Spin at ~ 125 × g for 5-7 minutes at room temperature, and discard the supernatant without disturbing the pellet.
6. Resuspend cell pellet with the appropriate volume of complete medium and transfer the cell suspension into a T25 culture flask.
7. Incubate the flask at 37°C, 5% CO2 incubator.
8. Split saturated culture 1:3-1:6 every 3 days; seed out at about 1-3 × 105 cells/mL.

Cell Freezing

1. Prepare the freezing medium (70% DMEM + 20% FBS + 10% DMSO) fresh immediately before use.
2. Keep the freezing medium on ice and label cryovials.
3. Transfer cells to a sterile, conical centrifuge tube, and count the cells.
4. Centrifuge the cells at 250×g for 5 minutes at room temperature and carefully aspirate off the medium.
5. Resuspend the cells at a density of at least 3×106 cells/mL in chilled freezing medium.
6. Aliquot 1 mL of the cell suspension into each cryovial.
7. Freeze cells in the CoolCell freezing container overnight in a -80°C freezer.
8. Transfer vials to liquid nitrogen for long-term storage

References

1. Zaballos, A., Varona, R., Guti¹Èrrez, J., Lind, P. & MÉÖrquez, G. Molecular cloning and RNA expression of two new human chemokine receptor-like genes. Biochemical and Biophysical Research Communications 227, 846ÿ853 (1996).
2. Kapur, N. et al. CCR6 expression in colon cancer is associated with advanced disease and supports epithelialto-mesenchymal transition. 114, 1343ÿ1351 (2016).
Please enable JavaScript in your browser to complete this form.