KC-4430

Ba/F3-BCR-ABL-M244V-Cell-Line

46463

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Background of Ba/F3-BCR-ABL-M244V-Cell-Line

The ABL1 gene is an oncogene located on the long arm of chromosome 9 at band q34 (9q34). Its gene product is a non-receptor tyrosine protein kinase. The BCR gene is located on the long arm of chromosome 22 at band q11 (22q11). Due to a translocation between the long arms of chromosomes 9 and 22, the ABL oncogene on chromosome 9 (9q34) and the BCR (B-cell receptor) gene on chromosome 22 (22q11) are recombined to form the BCR-ABL fusion gene. The BCR/ABL fusion gene is an anti-apoptotic gene with high tyrosine kinase activity, which causes excessive cell proliferation and leads to dysregulation of cellular control. 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-4430
Cell Line Name	Ba/F3-BCR-ABL-M244V-Cell-Line
Host Cell Line	Ba/F3
Description	Stable Ba/F3 clone expressing exogenous BCR-ABL gene bearing M244V mutation
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	Puromycin
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
In Vivo Validation	NA

Cell Line Generation

Ba/F3 BCR-ABL-M244V cell Line was generated using retrovirus vector expressing human BCR-ABL-M244V sequence.

Characterization

Figure 1: Characterization of BCR-ABL mutation in the Ba/F3 stable clone using PCR sequencing.

Figure 2: Characterization of IC50 (Half Inhibitory Concentration) in Ba/F3 BCR-ABL-M244V stable clone using GraphPad Prism software.

Cell Resuscitation

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

Cell Freezing

Prepare the freezing medium (70% RPMI-1640 + 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

Shibata N, Ohoka N, Hattori T, Naito M. Development of a Potent Protein Degrader against Oncogenic BCR-ABL Protein. Chem Pharm Bull (Tokyo). 2019;67(3):165-172. doi: 10.1248/cpb.c18-00703. PMID: 30827996.
Burke BA, Carroll M. BCR-ABL: a multi-faceted promoter of DNA mutation in chronic myelogeneous leukemia. Leukemia. 2010 Jun;24(6):1105-12. doi: 10.1038/leu.2010.67. Epub 2010 May 6. PMID: 20445577; PMCID: PMC4425294.

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.