KC-5557

K562-BCR-ABL-T315I-KI Cell Line

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Background of K562-BCR-ABL-T315I-KI Cell Line

A reciprocal translocation between chromosomes 22 and 9 produces the Philadelphia chromosome, which is often found in patients with chronic myelogenous leukemia. The chromosome 22 breakpoint for this translocation is located within the BCR gene. The translocation produces a fusion protein which is encoded by sequence from both BCR and ABL, the gene at the chromosome 9 breakpoint. BCR-ABL and its mutants can promote and maintain the malignant behavior of the cancer cells. The identification of BCR-ABL as a driver gene has led to the rapid development of anticancer therapeutics agents, including Imatinib, Dasatinib, Ponatinib and Nilotinib.

Specifications

Catalog Number	KC-5557
Cell Line Name	K562-BCR-ABL-T315I-KI Cell Line
Clone Number	1B2
Host Cell Line	K562
Description	Stable K562 clone expressing endogenous BCR-ABL gene bearing T315I mutations, No.1B2
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	RPMI1640+20% FBS+10% DMSO
Propagation Medium	RPMI1640+10% FBS
Selection Marker	NA
Morphology	hematopoietic
Subculture	Split saturated culture 1:5-1:8 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 24 hours
Mycoplasma Status	Negative

Cell Line Generation

K562-BCR-ABL-T315I-KI cell line was generated using the CRISPR method.

Characterization

Figure 1: Characterization of K562-BCR-ABL-T315I-KI cell line stable clone using PCR sequencing.

Figure 2: Characterization of K562-BCR-ABL-T315I-KI cell line stable clone using RT-PCR sequencing.

Figure 3: Characterization of Dose-response curves and IC50 values for K562 and K562-BCR-ABL-T315I-KI cells treated with Dasatinib,Ponatinib,Imatinib,Nilotinib and Asciminib over 3 days.

Cell Resuscitation

Prewarm culture medium (RPMI1640 + 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 1:5-1:8 every 2-3 days; seed out at about 1-3 ×10⁵ cells/mL.

Cell Freezing

Prepare the freezing medium (70% RPMI1640 + 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

Weisberg, E., Manley, P. W., Cowan-Jacob, S. W., Hochhaus, A. & Griffin, J. D. Second generation inhibitors of BCR-ABL for the treatment of imatinib-resistant chronic myeloid leukaemia. Nat Rev Cancer 7, 345ÿ356 (2007).

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.