KC-5948

CHOK1-STEAP2-Middle Cell Line

63016

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Background of CHOK1-STEAP2-Middle Cell Line

STEAP2 (Six-Transmembrane Epithelial Antigen of the Prostate 2), also known as STAMP1, PCANAP1, IPCA1, or PUMPCn, is a member of the STEAP family of metalloreductases. It is highly expressed in the prostate, with lower expression in the heart, brain, kidney, and pancreas. In cancer, STEAP2 is significantly upregulated in prostate cancer, including metastatic and hormone-refractory forms, making it a promising tumor-associated antigen for targeted therapy. Drug development has progressed to clinical trials, with AZD0516, an antibody-drug conjugate (ADC) targeting STEAP2, currently being evaluated for prostate cancer treatment.

Specifications

Catalog Number	KC-5948
Cell Line Name	CHOK1-STEAP2-Middle Cell Line
NCBI/UniProt Accession Number	NM_152999.3
Clone Number	5-33#
Host Cell Line	CHOK1 cell line
Description	Stable CHOK1 cell line expressing exogenous human STEAP2 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 30 hours
Mycoplasma Status	Negative
In Vivo Validation	NA

Cell Line Generation

CHOK1-STEAP2-Middle cell line was generated using a lentiviral vector expressing the human STEAP2 sequence.

Characterization

Figure 1: Characterization of STEAP2 overexpression in the CHOK1-STEAP2-Middle stable clone using FACS.

Figure 2: Characterization of STEAP2 in the CHOK1-STEAP2-Middle 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. 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.
2. 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.
3. 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.
4. Gomes, Inês M et al. “STEAP proteins: from structure to applications in cancer therapy.” Molecular cancer research : MCR vol. 10,5 (2012): 573-87. doi:10.1158/1541-7786.MCR-11-0281.