B-hROR1 CT26.WT

Basic Information

Description

• Origin: The CT26.WT cell line is derived from BALB/c murine colon adenocarcinoma cells. The cell line is a commonly used murine model for colorectal carcinoma.
• Background Information: The ROR1 encodes a receptor tyrosine kinase that has been implicated in nervous system development, specifically in the maintenance of neural progenitor cell fate, neurite extension and synapse formation. The encoded protein, likely a pseudokinase that lacks catalytic activity, may also regulate adipogenesis.
• Gene targeting strategy: The mouse Ror1 gene was replaced by human ROR1 coding sequence in B-hROR1 CT26.WT cells. Human ROR1 is highly expressed on the surface of B-hROR1 CT26.WT cells.
• Tumorigenicity: Confirmed in BALB/c mice.
• Application: B-hROR1 CT26.WT tumor models can be used for preclinical evaluation of monoclonal antibody drugs and bispecific antibody drugs targeting human ROR1.
• Notes:

1. Inoculated cell lines can be suspended with DMEM stock solution.
2. Before implementing the project, it is recommended to perform tumor growth experiments. The recommended cell inoculation amount is between 1E5-1E6.
3. In the experiment, it is necessary to ensure that the number of animals inoculated subcutaneously is at least 1.6 times the actual grouping number.

Protein expression analysis

ROR1 expression analysis in B-hROR1 CT26.WT cells by flow cytometry. Single cell suspensions from wild-type CT26.WT and B-hROR1 CT26.WT #2-H03 cultures were stained with species-specific anti-ROR1 antibody (Biolegend, 357804). Human ROR1 was detected on the surface of B-hROR1 CT26.WT cells but not wild-type CT26.WT cells.

Tumor growth curve & Body weight changes

Subcutaneous tumor growth of B-hROR1 CT26.WT. B-hROR1 CT26.WT (1×10⁵) and wild-type CT26.WT cells (1×10⁵) were subcutaneously implanted into BALB/c mice (female, 7-week-old, n=6). Tumor volume and body weight were measured three times a week. (A) Average tumor volume. (B) Body weight. Volume was expressed in mm³ using the formula: V=0.5 × long diameter × short diameter². Results indicate that B-hROR1 CT26.WT cells were able to establish tumors in vivo and can be used for efficacy studies. Values are expressed as mean ± SEM.

Protein expression analysis of tumor tissue

ROR1 expression evaluated on B-hROR1 CT26.WT tumor cells by flow cytometry. B-hROR1 CT26.WT cells were subcutaneously transplanted into BALB/c mice (n=6). Upon conclusion of the experiment, tumor cells were harvested and assessed for human ROR1 expression by flow cytometry (Biolegend, 357804). As shown, human ROR1 was highly expressed on the surface of tumor cells. Therefore, B-hROR1 CT26.WT cells can be used for in vivo efficacy studies evaluating novel ROR1 therapeutics.