KC-5635

CT26-KRAS-V8M-G12C-KI-Abcb1a-Abcb1b-KO Cell Line

57875

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Background of CT26-KRAS-V8M-G12C-KI-Abcb1a-Abcb1b-KO Cell Line

ABCB1b (ATP-binding cassette sub-family B member 1b), an isoform of the multidrug resistance protein 1 (MDR1), is encoded by the ABCB1 gene located on chromosome 7q21.12. It functions as a pivotal ATP-dependent efflux transporter highly expressed in excretory organs and blood-tissue barriers, such as the liver, kidney, and intestine. ABCB1b confers resistance to a broad spectrum of chemotherapeutic agents, including taxanes, anthracyclines, and vinca alkaloids, by actively pumping these substrates out of cancer cells. This efflux activity significantly reduces intracellular drug accumulation, linking ABCB1b overexpression to multidrug resistance (MDR) in various malignancies and posing a major obstacle to successful cancer chemotherapy.

Specifications

Catalog Number	KC-5635
Cell Line Name	CT26-KRAS-V8M-G12C-KI-Abcb1a-Abcb1b-KO Cell Line
Clone Number	1B4
Host Cell Line	CT26-KRAS-G12C-KI-Abcb1a-KO
Description	Stable CT26 clone with mouse Abca1b gene knockout.
Quantity	Two vials 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	Epithelial
Subculture	Split saturated culture 1:4-1:5 every 1-2 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

CT26-KRAS-G12C-KI-Abcb1a-Abcb1b-KO-1B4 cell line was generated using the CRISPR method.

Characterization

Figure 1: Characterization of CT26-KRAS-G12C-KI-Abcb1a-Abcb1b-KO cell line stable clone using PCR sequencing.

Figure 2: Characterization of CT26-KRAS-G12C-KI-Abcb1a-Abcb1b-KO cell line stable clone using RT-PCR sequencing.

Figure 3: Characterization of dose-response curves for KRAS inhibitors on CT26-KRAS-G12C-KI-Abcb1a-Abcb1b-KO cell line stable clone.

Figure 4:Characterization of CT26-KRAS-G12C-KI-Abcb1a-Abcb1b-KO cell line stable clone using western blot.

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 1:4-1:5 every 1-2 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

Chen, Z., Shi, T., Zhang, L., Zhu, P., Deng, M., Huang, C., Hu, T., Jiang, L., & Li, J. Mammalian drug efflux transporters of the ATP binding cassette (ABC) family in multidrug resistance: A review of the past decade. Cancer Letters 2016, 370 (1), 153–164. https://doi.org/10.1016/j.canlet.2015.10.010.