KC-1422

293T-FAP-Cell-Line

22109

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Background of 293T-FAP-Cell-Line

Fibroblast activation protein alpha (FAP) also named as prolyl endopeptidase FAP is a membrane-bound gelatinase, the expression of which is very low human tissue under physiological condition, and is high inactivated stromal fibroblasts of many human carcinoma.

Specifications

Catalog Number	KC-1422
Cell Line Name	293T-FAP-Cell-Line
Host Cell Line	293T
Description	Stable in culture over a minimum of 10 passages
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 1:4-1:8 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	Yes

Cell Line Generation

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

Characterization

Figure1: Characterization of FAP overexpression in 293T stable clones using FACS.

Cell Resuscitation

Prewarm culture medium (DMEM supplemented with 10% FBS and 1μg/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% 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

Rettig WJ, Su SL, Fortunato SR, Scanlan MJ, Raj BK, Garin-Chesa P, Healey JH, Old LJ (August 1994). Fibroblast
activation protein: purification, epitope mapping and induction by growth factors. International Journal of
Cancer. 58 (3): 385–92.
Garin-Chesa P, Old LJ, Rettig WJ (September 1990). Cell surface glycoprotein of reactive stromal fibroblasts as
a potential antibody target in human epithelial cancers. Proceedings of the National Academy of Sciences of
the United States of America. 87 (18): 7235–9.