kc-2983
PC3-PSMA-Cell-Line
Background of PC3-PSMA-Cell-Line
PSMA, also known as FOLH1, is a type II transmembrane glycoprotein belonging to the M28 peptidase family, which
is a glutamate-preferring carboxy-peptidase. PSMA is highly expressed in the 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-2983 |
|---|---|
| Cell Line Name | PC3-PSMA-Cell-Line |
| Host Cell Line | PC3 |
| Description | Stable PC3 cell line expressing exogenous human PSMA 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 | Ham's F12K + 20% FBS + 10% DMSO |
| Propagation Medium | Ham's F12K + 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 1-2 × 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
PC3 human PSMA cell line was generated using a lentiviral vector expressing the human PSMA sequence.
Characterization
Figure1: Characterization of human PSMA overexpression in PC3 human PSMA stable clones using FACS.
Figure2: Characterization of human PSMA expression in PC3 cells using FACS.
Figure3: Characterization of PC3 human PSMA stable clone using PCR sequencing.
Cell Resuscitation
- Prewarm culture medium (Ham's F12K 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 1-2 × 105 cells/mL.
Cell Freezing
- Prepare the freezing medium (Ham's F12K + 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
- Prostate-Specific Membrane Antigen (PSMA) Theranostics for Treatment of Oligometastatic Prostate Cancer. PMID: 34829977 PMCID: PMC8621856 DOI: 10.3390/ijms222212095.