KC-5547

293T-mouse-PSMA Cell Line

63929

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Background of 293T-mouse-PSMA Cell Line

FOLH1, also named as prostate-specific membrane antigen (PSMA), is enzyme protein and mainly expressed in prostate epithelium, the proximal tubules of the kidney, the jejunal brush border of the small intestine and ganglia of the nervous system. FOLH1 is overexpressed in human prostate cancer and might be a potential target for tumor therapy. Diseases associated with FOLH1 include Prostate Disease and Hyperhomocysteinemia.

Specifications

Catalog Number	KC-5547
Cell Line Name	293T-mouse-PSMA Cell Line
NCBI/UniProt Accession Number	NM_016770.3
Clone Number	2#
Host Cell Line	293T
Description	Stable 293T cell line expressing exogenous mouse PSMA gene
Quantity	One vial 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
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-mouse-PSMA cell line was generated using a lentiviral vector expressing the mouse PSMA sequence.

Characterization

Figure 1: Characterization of mouse PSMA overexpression in the 293T-mouse-PSMA stable clone using qPCR.

Figure 2: Characterization of mouse PSMA in the 293T-mouse-PSMA stable clone using PCR sequencing.

Cell Resuscitation

Pre-warm complete culture medium (basal medium and 10% FBS) in a 37°C water bath.
Rapidly thaw the cryovial in a 37°C water bath for 1-2 minutes with gentle agitation.
Transfer the vial to a biosafety cabinet, and disinfect the exterior with 70% ethanol.
Aseptically transfer the cell suspension dropwise into a sterile centrifuge tube containing 9.0 mL of pre-warmed complete medium.
Centrifuge at approximately 125 × g for 5–7 minutes at room temperature, carefully aspirate the supernatant without disturbing the cell pellet.
Gently resuspend the pellet in an appropriate volume of complete medium and transfer the suspension into a T25 flask.
Incubate the flask in a 37°C in a humidified 5% CO₂ incubator.
Assess cell viability and morphology after 24 hours. If cells appear healthy, replace the medium with fresh medium supplemented with the appropriate selective antibiotic.
Subculture the cells at a ratio of 1:4-1:8 every 2-3 days upon reaching 80%–90% confluency.

Cell Freezing

Prepare the freezing medium (70% basal medium, 20% FBS and 10% DMSO) freshly before use.
Pre-chill the freezing medium on ice and label the cryovials accordingly.
Transfer the cell suspension to a sterile conical tube and perform a cell count to determine total viability and density.
Centrifuge the cells at 250×g for 5 minutes at room temperature; carefully aspirate the supernatant.
Gently resuspend the cell pellet in chilled freezing medium, ensuring a minimum cell density of 3×10⁶ cells/mL.
Aliquot 1 mL of the cell suspension into each pre-labeled cryovial.
Place the cryovials into a CoolCell® container and store at -80°C overnight for controlled-rate cooling.
Transfer the cryovials to the liquid nitrogen for long-term storage the following day.

References

1. Farolfi A, Calderoni L, Mattana F, Mei R, Telo S, Fanti S, Castellucci P. Current and Emerging Clinical Applications of PSMA PET Diagnostic Imaging for Prostate Cancer. J Nucl Med. 2021 May 10;62(5):596-604. doi: 10.2967/jnumed.120.257238. Epub 2021 Mar 12. PMID: 33712536.
2. 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. Mapping, genomic organization and promoter analysis of the human prostate-specific membrane antigen gene. Biochim Biophys Acta. 1998 Nov 26;1443(1-2):113-27. doi: 10.1016/s0167-4781(98)00200-0. PMID: 9838072.
3. Bahler CD, Johnson MM, Davicioni E, Zhang J, Cheng L, Green MA, Koch MO. Predictors of Prostate-specific Membrane Antigen (PSMA/FOLH1) Expression in a Genomic Database. Urology. 2020 Oct;144:117-122. doi: 10.1016/j.urology.2020.06.025. Epub 2020 Jun 30. PMID: 32619596.
4. 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): 227–30.

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.