KC-3362

SK-OV-3-ERBB3 Cell Line

26633

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Background of SK-OV-3-ERBB3 Cell Line

ERBB3, also known as Her3, is a member of the epidermal growth factor receptor family of receptor tyrosine kinases. This membrane-bound glycoprotein has a neuregulin binding domain but has not an active kinase domain. It therefore can bind the ligand but cannot mediate the intracellular signal transduction through protein phosphorylation. However, it does form heterodimers with ErbB2 or other EGFR members responsible for tyrosine phosphorylation to give a receptor complex and initiate the related pathway, which lead to cell proliferation or differentiation. Overexpression of this protein has been reported in numerous cancers, including prostate, bladder, and breast tumors. This protein has different isoforms derived from alternative splicing variants, and among which, the secreted isoform lacking the intermembrane region modulates the activity of membrane-bound form.

Specifications

Catalog Number	KC-3362
Cell Line Name	SK-OV-3-ERBB3 Cell Line
NCBI/UniProt Accession Number	NM_001982.4
Clone Number	6#
Host Cell Line	SK-OV-3
Description	Stable SK-OV-3 cell line expressing exogenous ERBB3 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	DMEM+10% FBS +1μg/mL Puromycin
Selection Marker	Puromycin
Morphology	Epithelial
Subculture	Split saturated culture 1:3-1:6 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 40 hours
Mycoplasma Status	Negative
In Vivo Validation	Yes

Cell Line Generation

SK-OV-3-ERBB3 cell line was generated using lentivirus expressing ERBB3 sequence.

Characterization

Figure 1. Characterization of ERBB3 over-expression in the SK-OV-3-ERBB3 stable clone using FACS.

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 1:3-1:6 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

Hafeez U, Parslow AC, Gan HK, Scott AM. New insights into ErbB3 function and therapeutic targeting in cancer. Expert Rev Anticancer Ther. 2020 Dec;20(12):1057-1074. doi: 10.1080/14737140.2020.1829485. Epub 2020 Oct 26. PMID: 32981377.
Singer, E. etal., 2001, J. Biol. Chem. 276 (47): 44266–74.
Kraus,M.H.etal.,1989,Proc.Natl.Acad.Sci.86:9193-9197.

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.