KC-1700

293T-CCR5-Cell-Line

22643

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Background of 293T-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-1700
Cell Line Name	293T-CCR5-Cell-Line
Host Cell Line	Human HEK293T cell line
Description	Stable 293T cell line expressing exogenous human CCR5 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% DMEM+20% FBS+10% DMSO
Propagation Medium	DMEM+10%FBS+150µg/mL Hygromycin B
Selection Marker	HygromycinB
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
In Vivo Validation	NA

Cell Line Generation

293T human CCR5 cell line was generated using a lentiviral vector expressing the human CCR5 sequence.

Characterization

Cell Resuscitation

Prewarm culture medium (DMEM + 10% FBS + 150µg/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 1:3-1:6 every 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.