KC-4734

CHOK1-STEAP3 Cell Line

62526

Home » CHOK1-STEAP3 Cell Line

Background of CHOK1-STEAP3 Cell Line

STEAP3 (Six-Transmembrane Epithelial Antigen of the Prostate 3), also known as STMP3 and TSAP6 (Tumor Suppressor-Activated Pathway 6), is a metalloreductase enzyme that belongs to the STEAP family of proteins. It is primarily expressed in prostate tissue, hematopoietic stem cells, and erythroid precursors, where it plays a critical role in iron homeostasis by reducing ferric iron to ferrous iron for transferrin-mediated uptake. In cancer, STEAP3 is highly expressed in prostate cancer and has been associated with poor prognosis, tumor progression, and therapy resistance. It also functions in exosome biogenesis and has been implicated in chemoresistance mechanisms. As a tumor-associated antigen, STEAP3 is emerging as a promising target for cancer immunotherapy, with ongoing preclinical development of STEAP3-targeted antibodies, vaccines, and CAR-T cell therapies for prostate and other solid tumors.

Specifications

Catalog Number	KC-4734
Cell Line Name	CHOK1-STEAP3 Cell Line
NCBI/UniProt Accession Number	NM_182915.3
Clone Number	4#
Host Cell Line	CHOK1 cell line
Description	Stable CHOK1 cell line expressing exogenous human STEAP3 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% basal medium+20% FBS+10% DMSO
Propagation Medium	RPMI1640+10% FBS+10μg/mL Puromycin
Selection Marker	Puromycin
Morphology	Epithelial-like
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 28 hours
Mycoplasma Status	Negative
In Vivo Validation	NA

Cell Line Generation

CHOK1-STEAP3 cell line was generated using a lentiviral vector expressing the human STEAP3 sequence.

Characterization

Figure 1: Characterization of STEAP3 overexpression in the CHOK1-STEAP3 stable clone using qPCR.

Figure 2: Characterization of STEAP3 in the CHOK1-STEAP3 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. Graham, Ross-M et al. “Liver iron transport.” World journal of gastroenterology vol. 13,35 (2007): 4725-36. doi:10.3748/wjg.v13.i35.4725.
2. Cai, Qiaomei et al. “STEAP Proteins: Roles in disease biology and potential for therapeutic intervention.” International journal of biological macromolecules vol. 309,Pt 1 (2025): 142797. doi:10.1016/j.ijbiomac.2025.142797.
3. Xu, Michael et al. “STEAP1-4 (Six-Transmembrane Epithelial Antigen of the Prostate 1-4) and Their Clinical Implications for Prostate Cancer.” Cancers vol. 14,16 4034. 20 Aug. 2022, doi:10.3390/cancers14164034.
4. Sikkeland, Jørgen et al. “STAMPing at the crossroads of normal physiology and disease states.” Molecular and cellular endocrinology vol. 425 (2016): 26-36. doi:10.1016/j.mce.2016.02.013.