KC-2079-DW

CT26-ROR1 Cell Line

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Background of CT26-ROR1 Cell Line

ROR1, also named as NTRKR1, is a transmembrane protein belong to the receptor tyrosine kinase-like orphan receptor (ROR) family. ROR1 modulates neurite growth in the central nervous system. ROR1 has recently found to be overexpressed on cancer stem cell, which play a function role in promoting cancer cell migration, invasion or spheroid formation.

Specifications

Catalog Number	KC-2079-DW
Cell Line Name	CT26-ROR1 Cell Line
Clone Number	NA
Host Cell Line	Mouse CT26 Cell Line
Description	Stable CT26 clone expressing exogenous human ROR1 gene
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 + 10μg/mL Puromycin
Selection Marker	Puromycin
Morphology	Fibroblast
Subculture	Split saturated culture 1:2-1:4 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
In Vivo Validation	Yes

Cell Line Generation

CT26 ROR1 cell line was generated using a lentiviral vector expressing the human ROR1 sequence.

Characterization

Figure 1: Characterization of human ROR1 overexpression CT26 using FACS.

Figure 2: Characterization of human ROR1overexpression CT26 ROR1 stable clone using FACS.

Cell Resuscitation

1. Prewarm culture medium (RPMI1640 supplemented with 10% FBS and 10μg/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 1:2-1:4 every 2-3 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.Kipps TJ. ROR1: an orphan becomes apparent. Blood. 2022 Oct 6;140(14):1583-1591. doi: 10.1182/blood.2021014760. PMID: 35580162; PMCID: PMC10653015.
2. Osorio-Rodríguez DA, Camacho BA, Ramírez-Segura C. Anti-ROR1 CAR-T cells: Architecture and performance. Front Med (Lausanne). 2023 Feb 17;10:1121020. doi: 10.3389/fmed.2023.1121020. PMID: 36873868; PMCID: PMC9981679.
3. Liu D, Gunther K, Enriquez LA, Daniels B, O\'Mara TA, Tang K, Spurdle AB, Ford CE. ROR1 is upregulated in endometrial cancer and represents a novel therapeutic target. Sci Rep. 2020 Aug 17;10(1):13906. doi: 10.1038/s41598-020-70924-z. Erratum in: Sci Rep. 2022 Jun 28;12(1):10918. doi: 10.1038/s41598-022-15043-7. PMID: 32807831; PMCID: PMC7431863.

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.