KC-6503

B16/F10-STEAP2-ECD Cell Line

64116

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Background of B16/F10-STEAP2-ECD Cell Line

STEAP2 (STEAP2 Metalloreductase) is a Protein Coding gene. This gene is a member of the STEAP family and encodes a multi-pass membrane protein that localizes to the Golgi complex, the plasma membrane, and the vesicular tubular structures in the cytosol. It is highly over-expressed in various types of cancer and it up-regulation hindered cellular proliferation, invasion and metastasis abilities by inhibiting EMT process and suppressing PI3K/AKT/mTOR signaling pathway. Diseases associated with STEAP2 include Hepatic Adenomas, Familial and Prostate Cancer.

Specifications

Catalog Number	KC-6503
Cell Line Name	B16/F10-STEAP2-ECD Cell Line
NCBI/UniProt Accession Number	NM_152999.3
Clone Number	2#
Host Cell Line	B16/F10
Description	Stable B16/F10 cell line expressing exogenous STEAP2 Extracellular domain 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	90% DMEM+10% FBS
Selection Marker	NA
Morphology	mixture of spindle-shaped and epithelial-like cells
Subculture	Split saturated culture 1:4 to 1:8 every 2-3 days; seed out at about 1-3 × 10⁵ cells/mL
Incubation	37 °C with 5% CO₂
Storage	Liquid nitrogen immediately upon receiving
Doubling Time	Approximately 20 hours
Mycoplasma Status	Negative

Cell Line Generation

B16/F10-STEAP2-ECD Cell Line was generated using a lentiviral vector expressing the STEAP2 Extracellular domain sequence.

Characterization

Figure 1.Characterization of STEAP2 Extracellular domain overexpression in the B16/F10-STEAP2-ECD stable clone using PCR sequence.

Figure 2:Characterization of STEAP2 Extracellular domain overexpression in the B16/F10-STEAP2-ECD stable clone using FACS.

Cell Resuscitation

Pre-warm complete culture medium (90% DMEM+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 to 1:8 every 2-3 days upon reaching 80%–90% confluency.

Cell Freezing

Prepare the freezing medium (70% DMEM+20% FBS+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

Torrez CZ, Easley A, Bouamar H, Zheng G, Gu X, Yang J, Chiu YC, Chen Y, Halff GA, Cigarroa FG, Sun LZ. STEAP2 promotes hepatocellular carcinoma progression via increased copper levels and stress-activated MAP kinase activity. Sci Rep. 2024 Jun 3;14(1):12753.
Yang Q, Ji G, Li J. STEAP2 is down-regulated in breast cancer tissue and suppresses PI3K/AKT signaling and breast cancer cell invasion in vitro and in vivo. Cancer Biol Ther. 2020;21(3):278-291.

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.