KC-1551

293T-FOLR1/PSMA-Cell-Line

22351

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Background of 293T-FOLR1/PSMA-Cell-Line

FOLR1 & FOLH, also known as FOLR & PSMA. FOLR1 is a protein that belongs to a member of the folate receptor family. FOLR1 binds to folic acid and its derivatives, which becomes crucial during fetal development. FOLR1, due to its high expression in some tumors, is an attractive therapeutic target for the development of novel anti-cancer agents in order to limit toxic side effects on off-target tissues. FOLH is a type II transmembrane glycoprotein belonging to the M28 peptidase family, a glutamate-preferring carboxy-peptidase. PSMA is highly expressed in prostate secretory-acinar epithelium, in some benign extraprostatic epithelial cells from breast, duodenum, and kidney tissues, and prostate cancer. In the prostate, the protein is up-regulated in cancerous cells and is used as an effective diagnostic and prognostic indicator of prostate cancer.

Specifications

Catalog Number	KC-1551
Cell Line Name	293T-FOLR1/PSMA-Cell-Line
Host Cell Line	Human HEK293T cell line
Description	Stable HEK293T cell line expressing exogenous human FOLR1 FOLH1 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+150µg/mL Hygromycin B
Selection Marker	Puromycin, 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 FOLR1 FOLH1 Cell Line was generated using a lentiviral vector expressing the human FOLR1 FOLH1 sequence.

Characterization

Cell Resuscitation

Prewarm culture medium (DMEM + 10% FBS + 1µg/mL Puromycin + 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

O'Keefe DS, Su SL, Bacich DJ, Horiguchi Y, Luo Y, Powell CT, Zandvliet D, Russell PJ, Molloy PL, Nowak NJ, Shows TB, Mullins C, Vonder Haar RA, Fair WR, Heston WD (November 1998). "Mapping, genomic organization and promoter analysis of the human prostate-specific membrane antigen gene". Biochim. Biophys. Acta. 1443 (1ÿ 2): 113ÿ27.
Israeli RS, Powell CT, Fair WR, Heston WD (January 1993). "Molecular cloning of a complementary DNA encoding a prostate-specific membrane antigen". Cancer Res. 53 (2): 227ÿ30.