KC-5882

CHOK1-mouse-CXCR5 Cell Line

59869

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Background of CHOK1-mouse-CXCR5 Cell Line

This gene encodes a multi-pass membrane protein that belongs to the CXC chemokine receptor family. It is expressed in mature B-cells and Burkitt's lymphoma. This cytokine receptor binds to B-lymphocyte chemoattractant (BLC), and is involved in B-cell migration into B-cell follicles of spleen and Peyer patches. Alternatively spliced transcript variants encoding different isoforms have been described for this gene.

Specifications

Catalog Number	KC-5882
Cell Line Name	CHOK1-mouse-CXCR5 Cell Line
NCBI/UniProt Accession Number	NM_007551.3
Clone Number	NA
Host Cell Line	CHOK1
Description	Stable CHOK1 cell line expressing exogenous mouse-CXCR5 gene
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 + 10μg/mL Puromycin
Selection Marker	Puromycin
Morphology	Epithelial
Subculture	Split saturated culture 1:4-1:8 every 2-3 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

CHOK1-mouse-CXCR5 Cell Line was generated using a lentiviral vector expressing the mouse-CXCR5 sequence.

Characterization

Figure 1: Characterization of mouse-CXCR5 overexpression in the CHOK1-mouse-CXCR5 stable clone using FACS.

Figure 2: Characterization of mouse-CXCR5 overexpression in the CHOK1-mouse-CXCR5 stable clone using PCR.

Cell Resuscitation

Prewarm culture medium (RPMI1640 + 10% FBS + 10μg/mL Puromycin)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 1:4-1:8 every 2-3 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

Zhao S, Fang F, Tang X, Dou J, Wang W, Zheng X, Sun L, Zhang A. An in-depth analysis identifies two new independent signals in 11q23.3 associated with vitiligo in the Chinese Han population. J Dermatol Sci. 2017 Oct;88(1):103-109. doi: 10.1016/j.jdermsci.2017.05.001. Epub 2017 May 3. PMID: 28551095.
Meijer J, Zeelenberg IS, Sipos B, Roos E. The CXCR5 chemokine receptor is expressed by carcinoma cells and promotes growth of colon carcinoma in the liver. Cancer Res. 2006 Oct 1;66(19):9576-82. doi: 10.1158/0008-5472.CAN-06-1507. PMID: 17018614.