KC-3771

293T-FPR1 Cell Line

34748

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Background of 293T-FPR1 Cell Line

This gene encodes a G protein-coupled receptor of mammalian phagocytic cells that is a member of the G-protein coupled receptor 1 family. The protein mediates the response of phagocytic cells to invasion of the host by microorganisms and is important in host defense and inflammation.

Specifications

Catalog Number	KC-3771
Cell Line Name	293T-FPR1 Cell Line
Host Cell Line	293T
Description	Stable HEK293T clone expressing exogenous FPR1 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 + 1μg/mL Puromycin
Selection Marker	Puromycin
Morphology	Epithelial
Subculture	Split saturated culture 1:4-1:5 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 FPR1 cell line was generated using a lentiviral vector expressing the FPR1 sequence.

Characterization

Figure 1: Characterization of endogenous FPR1 expression in 293T cells using FACS.

Figure 2: Characterization of FPR1 overexpression in the 293T FPR1 stable clone using FACS.

Figure 3: Characterization of FPR1 in the 293T FPR1 stable clone using PCR sequencing.

Cell Resuscitation

1. Prewarm culture medium (DMEM supplemented with 10% FBS and 1μg/mL puromycin)in a 37°C water bath. 2. Thaw the frozen vial in a 37°C water bath for 1-2 minutes. 3. Transfer the vial into biosafety cabinet, and wipe the surface with 70% ethanol. 4. Unscrew the top of the vial and transfer the cell suspension gently into a sterile centrifuge tube containing 9.0mL complete culture medium. 5. Spin at ~ 125 × g for 5-7 minutes at room temperature, and discard the supernatant without disturbing the pellet. 6. Resuspend cell pellet with the appropriate volume of complete medium and transfer the cell suspension into a T25 culture flask. 7. Incubate the flask at 37°C, 5% CO₂ incubator. 8. Split saturated culture 1:4-1:5 every 2-3 days; seed out at about 1-3 × 10⁵ cells/mL.

Cell Freezing

1. Prepare the freezing medium (70% DMEM + 20% FBS + 10% DMSO) fresh immediately before use. 2. Keep the freezing medium on ice and label cryovials. 3. Transfer cells to a sterile, conical centrifuge tube, and count the cells. 4. Centrifuge the cells at 250×g for 5 minutes at room temperature and carefully aspirate off the medium. 5. Resuspend the cells at a density of at least 3×10⁶ cells/mL in chilled freezing medium. 6. Aliquot 1 mL of the cell suspension into each cryovial. 7. Freeze cells in the CoolCell freezing container overnight in a -80°C freezer. 8. Transfer vials to liquid nitrogen for long-term storage.

References

1. Zhu XL, Meng HX, Zhang L, Xu L, Chen ZB, Shi D, Feng XH. [Association of SNPs in N-formylpeptide receptor gene with susceptibility of aggressive periodontitis]. Beijing Da Xue Xue Bao Yi Xue Ban. 2009 Dec 18;41(6):664-8. Chinese. PMID: 20019777. 2. Kim MK, Min do S, Park YJ, Kim JH, Ryu SH, Bae YS. Expression and functional role of formyl peptide receptor in human bone marrow-derived mesenchymal stem cells. FEBS Lett. 2007 May 1;581(9):1917-22. doi: 10.1016/j.febslet.2007.03.078. Epub 2007 Apr 9. PMID: 17442310.