KC-6289

Eol-1-CCR6-KO cell line

61441

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Background of Eol-1-CCR6-KO cell line

The CC chemokine receptor 6 (CCR6), alternatively known as STRL22, resides on chromosome 6q27 and functions as a homologous receptor for CCL20. It is also characterized as a 7-transmembrane domain G-protein coupled receptor, primarily expressed in leukocytes, and binds to CCL20 to mediate leukocyte migration. In tissues, it is predominantly expressed in the lymph nodes, appendix, and spleen, while showing lower levels of expression in the thymus, testis, and small intestine .

Specifications

Catalog Number	KC-6289
Cell Line Name	Eol-1-CCR6-KO cell line
Clone Number	1A2
Host Cell Line	EOL1
Description	Stable Eol-1-CCR6-KO cell clone with human CCR6 gene knockout
Quantity	Two vials of frozen cells (≥2-10⁶/vial)
Stability	Stable in culture over a minimum of 10 passages
Application	Drug screening and biological assays
Freezing Medium	70% RPMI1640 + 20% FBS + 10% DMSO
Propagation Medium	RPMI1640 + 10% FBS
Selection Marker	NA
Morphology	Lymphoblast
Subculture	Split the saturated culture at a ratio of 1:3-1:4 every 3-4 days; seed out at about 1-3 x 10⁵ cells/mL
Incubation	37 °C with 5% CO₂
Storage	Liquid nitrogen immediately upon receiving
Doubling Time	Approximately 32 hours
Mycoplasma Status	Negative

Cell Line Generation

Eol-1-CCR6-KO cell line was generated using the CRISPR method.

Characterization

Figure 1: Characterization of Eol-1-CCR6-KO cell line stable clone using PCR sequencing.

Figure 2:Characterization of Eol-1-CCR6-KO cell line stable clone using RT-PCR sequencing.

Figure 3:Characterization of Eol-1-CCR6-KO cell line stable clone using FACS.

Cell Resuscitation

Prewarm culture medium (RPMI1640+10% FBS)in a 37°C water bath.
Thaw the frozen vial in a 37°C water bath for 1-2 minutes.
Transfer the vial into biosafety cabinet, and wipe the surface with 70% ethanol.
Unscrew the top of the vial and transfer the cell suspension gently into a sterile centrifuge tube containing 9.0mL complete culture medium.
Spin at ~ 125 × g for 5-7 minutes at room temperature, and discard the supernatant without disturbing the pellet.
Resuspend cell pellet with the appropriate volume of complete medium and transfer the cell suspension into a T25 culture flask.
Incubate the flask at 37°C, 5% CO₂ incubator.
Split saturated culture 1:3-1:4 every 3-4 days; seed out at about 1-3 × 10⁵ cells/mL.

Cell Freezing

Prepare the freezing medium (70% RPMI1640 + 20% FBS + 10% DMSO) fresh immediately before use.
Keep the freezing medium on ice and label cryovials.
Transfer cells to a sterile, conical centrifuge tube, and count the cells.
Centrifuge the cells at 250×g for 5 minutes at room temperature and carefully aspirate off the medium.
Resuspend the cells at a density of at least 3×10⁶ cells/mL in chilled freezing medium.
Aliquot 1 mL of the cell suspension into each cryovial.
Freeze cells in the CoolCell freezing container overnight in a -80°C freezer.
Transfer vials to liquid nitrogen for long-term storage.

References

Li Y-, Geng WL, Li CC, Wu JH, Gao F, Wang Y. Progress of CCL20-CCR6 in the airways: a promising new therapeutic target. J Inflamm (Lond). 2024 Dec 27;21(1):54. doi: 10.1186/s12950-024-00427-5. Erratum in: J Inflamm (Lond). 2025 Jan 13;22(1):2. doi: 10.1186/s12950-025-00428-y. PMID: 39731176; PMCID: PMC11681768.