KC-6573

Ba/F3-ERBB2-T798M-Cell-Line

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Background of Ba/F3-ERBB2-T798M-Cell-Line

ERBB2, also known as CD34, encodes a member of the epidermal growth factor (EGF) receptor tyrosine kinase family. This protein itself does not have a ligand binding domain, so it cannot bind to growth factors. However, it does tightly bind with other ligands in the EGF receptor family to form heterodimers, stabilizing ligand binding and enhancing activation of downstream signaling pathways mediated by kinases, such as those involving mitogen activated protein kinase and phosphatidylinositol 3-kinase. It is reported that the amplification and/or overexpression of this gene are found in many cancers, including breast cancer and ovarian tumors.
Ba/F3 cell, a murine interleukin-3 dependent pro-B cell line, is a popular system for exploring both kinases and their inhibitors, because some protein kinases can render the Ba/F3 cells to be depended on the activation of the kinases instead of IL-3 supplement, while their inhibitors can antagonize the kinase-dependent growth effects.

Specifications

Catalog Number	KC-6573
Cell Line Name	Ba/F3-ERBB2-T798M-Cell-Line
NCBI/UniProt Accession Number	NM_004448
Clone Number	4#
Host Cell Line	Ba/F3
Description	Stable Ba/F3 clone expressing exogenous ERBB2 bearing T798M amino acid mutations
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% RPMI1640+20% FBS+10% DMSO
Propagation Medium	RPMI1640+10%FBS
Selection Marker	NA
Morphology	Mostly single, round (some polymorph) cells in suspension
Subculture	Split saturated culture 1:10 every 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

Ba/F3-ERBB2-T798M-cell-Line was generated using retrovirus vector expressing human ERBB2-T798M sequence.

Characterization

Figure 1: Characterization of ERBB2-T798M in the Ba/F3-ERBB2-T798M stable clone using PCR sequencing.

Cell Resuscitation

1. Prewarm culture medium (RPMI1640 + 10% FBS)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 1:10 every 3 days; seed out at about 1-3 × 10⁵ cells/mL.

Cell Freezing

1. Prepare the freezing medium (70% RPMI1640 + 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. Idamakanti M, Bijjam RI, Kumar M, Mukkamalla SK. Long Durable Response With Trastuzumab Deruxtecan Monotherapy in a Triple-Negative Metastatic Breast Cancer Patient With Human Epidermal Growth Factor Receptor 2 Mutation: A Long-Term Follow-Up and Literature Review. J Med Cases. 2025 Jun 16;16(6):212-221. doi: 10.14740/jmc5136. PMID: 40642740; PMCID: PMC12239829.
2.Zheng Y, Shen G, Zhang C, Huo X, Xin Y, Fang Q, Guan Y, Zhao F, Ren D, Liu Z, Wang M, Zhao J. Efficacy of anti-HER2 drugs in the treatment of patients with HER2-mutated cancers: a systematic review and meta-analysis. Clin Exp Med. 2023 Nov;23(7):3205-3216. doi: 10.1007/s10238-023-01072-7. Epub 2023 Apr 30. PMID: 37120775.

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.