KC-4829

293T-STEAP3 Cell Line

59072

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Background of 293T-STEAP3 Cell Line

This gene encodes a multipass membrane protein that functions as an iron transporter. The encoded protein can reduce both iron (Fe3+) and copper (Cu2+) cations. This protein may mediate downstream responses to p53, including promoting apoptosis. Deficiency in this gene can cause anemia. Alternative splicing results in multiple transcript variants.

Specifications

Catalog Number	KC-4829
Cell Line Name	293T-STEAP3 Cell Line
Clone Number	1#
Host Cell Line	293T
Description	Stable 293T cell line expressing exogenous 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% DMEM + 20% FBS + 10% DMSO
Propagation Medium	DMEM + 10% FBS + 1μg/mL Puromycin
Selection Marker	Puromycin
Morphology	Epithelial
Subculture	Split saturated culture 1:4-1:5 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 30 hours
Mycoplasma Status	Negative

Cell Line Generation

293T-STEAP3 cell line was generated using a lentiviral vector expressing the STEAP3 sequence.

Characterization

Figure 1: Characterization of STEAP3 overexpressing in 293T-STEAP3 stable clone using PCR sequencing.

Figure 2: Characterization of STEAP3 overexpression in the 293T-STEAP3 stable clone using QPCR.

Cell Resuscitation

Prewarm culture medium (DMEM 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 1:4-1:5 every 2-3 days; seed out at about 1-3 × 10⁵ cells/mL.

Cell Freezing

Prepare the freezing medium (70% DMEM + 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

Liu D, Yi S, Zhang X, Fang P, Zheng C, Lin L, Cai R, Ye Y, Zhou Y, Liang Y, Cheng F, Zhang X, Zhou W, Mohandas N, An X, Xu X. Human STEAP3 mutations with no phenotypic red cell changes. Blood. 2016 Feb 25;127(8):1067-71. doi: 10.1182/blood-2015-09-670174. Epub 2015 Dec 16. PMID: 26675350; PMCID: PMC4768430.
Grunewald TG, Bach H, Cossarizza A, Matsumoto I. The STEAP protein family: versatile oxidoreductases and targets for cancer immunotherapy with overlapping and distinct cellular functions. Biol Cell. 2012 Nov;104(11):641-57. doi: 10.1111/boc.201200027. Epub 2012 Sep 24. PMID: 22804687.