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
NCBI/UniProt Accession Number	NM_001394783.1
Clone Number	2#
Host Cell Line	293T
Description	Stable 293T cell line expressing exogenous CCR5 gene.
Quantity	One vial of frozen cells (≥2×10⁶ cells)
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	Hygromycin B
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
In Vivo Validation	NA

Cell Line Generation

293T-CCR5 cell line was generated using lentivirus expressing CCR5 sequence.

Characterization

Figure 1. Characterization of CCR5 over-expression in the 293T-CCR5 stable clone using FACS.

Cell Resuscitation

Prewarm culture medium (DMEM supplemented with 10% FBS and 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 1:4-1:8 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

Hernanz-Falcón P, Rodríguez-Frade JM, Serrano A, Juan D, del Sol A, Soriano SF, Roncal F, Gómez L, Valencia A, Martínez-A C, Mellado M. Identification of amino acid residues crucial for chemokine receptor dimerization. Nat Immunol. 2004 Feb;5(2):216-23. doi: 10.1038/ni1027. Epub 2004 Jan 11. PMID: 14716309.

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.