KC-6038

LNcap-STEAP1-PSMA-KO cell line

60326

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Background of LNcap-STEAP1-PSMA-KO cell line

The protein encoded by the FOLH1 gene is commonly referred to as PSMA in the oncology community. However, neurological and metabolic studies describe FOLH1 as glutamate carboxypeptidase II (GCPII) or N-acetyl-L-aspartyl-L-glutamate peptidase I (NAALA-Dase I). Setting aside nomenclature differences, this enzyme — PSMA, GCPII or NAALADase I — functions as a transmembrane glycoprotein with both folate hydrolase and carboxypeptidase capabilities. This protein is predominantly expressed in the prostate gland, renal tissue and duodenum, where it has a vital role in the enzymatic processing of dietary folates. Structurally, PSMA consists of a 19-amino-acid intracellular portion, a 24-amino-acid transmembrane portion, and a 707-amino-acid extracellular portion.

Specifications

Catalog Number	KC-6038
Cell Line Name	LNcap-STEAP1-PSMA-KO cell line
Clone Number	3C2
Host Cell Line	LNcap-STEAP1-KO
Description	Stable LNcap-STEAP1-KO cell clone with human PSMA gene knockout
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% RPMI1640 + 20% FBS + 10% DMSO
Propagation Medium	RPMI1640 + 10% FBS
Selection Marker	NA
Morphology	Fibroblastoid cells growing as a monolayer
Subculture	Split the saturated culture at a ratio of 1:3-1:4 every 3-4 days; seed out at about 1-3 x 10⁵ cells/mL
Incubation	37 °C with 5% CO₂
Storage	Liquid nitrogen immediately upon receiving
Doubling Time	Approximately 36 hours
Mycoplasma Status	Negative

Cell Line Generation

LNcap-STEAP1-PSMA-KO cell line was generated using the CRISPR method.

Characterization

Figure 1: Characterization of LNcap-STEAP1-PSMA-KO cell line stable clone using PCR sequencing.

Figure 2:Characterization of LNcap-STEAP1-PSMA-KO cell line stable clone using RT-PCR sequencing.

Figure 3:Characterization of LNcap-STEAP1-PSMA-KO cell line stable clone using FACS.

Cell Resuscitation

Prewarm culture medium (RPMI1640+10% FBS)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 1:3-1:4 every 3-4 days; seed out at about 1-3 × 10⁵ cells/mL.

Cell Freezing

Prepare the freezing medium (70% RPMI1640 + 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

Bakht MK, Beltran H. Biological determinants of PSMA expression, regulation and heterogeneity in prostate cancer. Nat Rev Urol. 2025 Jan;22(1):26-45. doi: 10.1038/s41585-024-00900-z. Epub 2024 Jul 8. PMID: 38977769; PMCID: PMC11841200.