KC-3343

HCC827-ERBB3 Cell Line

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Background

ERBB3, also known as HER3, is a transmembrane protein belonging to epidermal growth factor receptor (ERBB) family of receptor tyrosine kinase. The kinase domain impaired ERBB3 can from active heterodimer with other member of ERBB family, this heterodimer, mostly with ERBB2 after binding with its ligand NRG1 or NRG2, play an important role in cancer growth, invasion, metastasis and chemotherapeutic resistant.

Specifications

Catalog Number:
KC-3343
Cell Line Name:
HCC827-ERBB3 Cell Line
Host Cell Line:
Description:
Stable HCC827 cell line expressing exogenous ERBB3 gene
Quantity:
One vial of frozen cells (5×106 per vial)
Stability:
Stable in culture over a minimum of 10 passages
Application:
Drug screening and biological assays
Freeze Medium:
70% RPMI1640+20% FBS+10% DMSO
Propagation Medium:
RPMI1640+10%FBS+2µg/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 30 hours
Mycoplasma Status:
Negative

Cell Line Generation

HCC827-human ERBB3 Cell Line was generated using a lentiviral vector expressing the ERBB3 sequence.

Characterization

Application

Cell Resuscitation

1. Prewarm culture medium (RPMI1640 + 10% FBS + 2µg/mL Puromycin) in a 37°C water bath.
2. Thaw the frozen vial in a 37°C water bath for 1-2 minutes.
3. Transfer the vial into biosafety cabinet, and wipe the surface with 70% ethanol.
4. Unscrew the top of the vial and transfer the cell suspension gently into a sterile centrifuge tube containing 9.0mL complete culture medium.
5. Spin at ~ 125 × g for 5-7 minutes at room temperature, and discard the supernatant without disturbing the pellet.
6. Resuspend cell pellet with the appropriate volume of complete medium and transfer the cell suspension into a T25 culture flask.
7. Incubate the flask at 37°C, 5% CO2 incubator.
8. Split saturated culture 1:3-1:6 every 2-3 days; seed out at about 1-3 × 105 cells/mL.

Cell Freezing

1. Prepare the freezing medium (70% RPMI-1640 + 20% FBS + 10% DMSO) fresh immediately before use.
2. Keep the freezing medium on ice and label cryovials.
3. Transfer cells to a sterile, conical centrifuge tube, and count the cells.
4. Centrifuge the cells at 250×g for 5 minutes at room temperature and carefully aspirate off the medium.
5. Resuspend the cells at a density of at least 3×106 cells/mL in chilled freezing medium.
6. Aliquot 1 mL of the cell suspension into each cryovial.
7. Freeze cells in the CoolCell freezing container overnight in a -80°C freezer.
8. Transfer vials to liquid nitrogen for long-term storage

References

1. Jaiswal, B. S. et al. Oncogenic ERBB3 Mutations in Human Cancers. Cancer Cell 23, 603ÿ617 (2013).
2. Baselga, J. & Swain, S. M. Novel anticancer targets: Revisiting ERBB2 and discovering ERBB3. Nat. Rev. Cancer 9, 463ÿ475 (2009).
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