KC-1715

CT26-CCR5-Cell-Line

22671

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Background of CT26-CCR5-Cell-Line

CCR5, also known as CD195, is a G protein-linked seven transmembrane domain receptor that belongs to the beta chemokine receptors family, which is a protein on the surface of white blood cells that is involved in the immune system. CCR5 is predominantly expressed on T cells, macrophages, dendritic cells, eosinophils, microglia and a subpopulation of either breast or prostate cancer cells. CCR5 inhibitors blocked the migration and metastasis of breast and prostate cancer cells that expressed CCR5, that may function as a new therapeutic target.

Specifications

Catalog Number	KC-1715
Cell Line Name	CT26-CCR5-Cell-Line
Host Cell Line	Mouse CT26 cell line
Description	Stable CT26 cell line expressing exogenous human CCR5 gene
Quantity	One vial 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% DMEM+20% FBS+10% DMSO
Propagation Medium	DMEM+10%FBS+600µg/mL Hygromycin B
Selection Marker	HygromycinB
Morphology	Fibroblast
Subculture	Split saturated culture 1:4-1:10 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 24 hours
Mycoplasma Status	Negative
In Vivo Validation	NA

Cell Line Generation

CT26 human CCR5 cell line was generated using a lentiviral vector expressing the human CCR5 sequence.

Characterization

Cell Resuscitation

Prewarm culture medium (DMEM + 10%FBS + 600µg/mL Hygromycin B)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 1:4-1:10 every 2-3 days; seed out at about 1-3 × 10⁵ cells/mL.

Cell Freezing

Prepare the freezing medium (70% DMEM + 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

Aldinucci D, Borghese C, Casagrande N. The CCL5/CCR5 Axis in Cancer Progression. Cancers (Basel). 2020 Jul 2;12(7):1765.
Hemmatazad H, Berger MD. CCR5 is a potential therapeutic target for cancer. Expert Opin Ther Targets. 2021 Apr;25(4):311-327.
Prathipati PK, Mandal S, Destache CJ. A review of CCR5 antibodies against HIV: current and future aspects. Ther Deliv. 2019 Feb;10(2):107-113.

Use License Agreement

Research Use Only.

Not for use in diagnostic procedures or therapeutic applications.

Redistribution of the cell line or its derivatives is prohibited without prior written permission from Kyinno Biotechnology.