At present, cell therapy is mostly used in the field of oncology, in which it is necessary to construct stable tumor cell models and animal models to evaluate the therapeutic effect. There are many factors that affect the effectiveness of CAR-T cells, such as transfection efficiency, culture conditions, and cell types. In vitro cell models provide a variety of detection methods for evaluating the effectiveness and specificity of CAR-T therapy. For in vivo evaluation models, it is required that the model have a corresponding immune environment to avoid causing CAR T-cell rejection, and the experimental animals must have a stable living condition. In addition, a mouse model of homologous tumor transplantation can also be used to verify the mechanism and treatment principles of CAR-T.
Cyagen has been committed to the establishment of gene-edited cells and animal models. We have an experienced team of experts and a mature technology platform that provides cell models and animal models for the regulation of target gene expression. For CAR-T and cell therapy research, we have established catalogs with thousands of cell and mouse models for tumor immune research, and provide preclinical in vivo/in vitro pharmacology evaluation services for domestic and foreign pharmaceutical companies, biological companies, scientific research institutes, and hospitals. For example, C-NKG mice are a severe combined immunodeficiency (SCID) model developed by Cyagen, which are without murine T/B/NK cells and have defective myeloid components. C-NKG mice can be used to construct various CDX models and reconstruction of the human immune system in a mouse model.
In vitro evaluation models
Services：
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Type |
Model |
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Cell Model |
Tumor cells expressing CAR-T targets |
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Luciferase labeled tumor cells |
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CAR-T cell construction |
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Overexpression stable transgenic cell Line services |
Case Study
1. Construction of CD19-293T cell line
A 293T cell line with stable expression of CD19 was constructed by lentivirus infection, and the high expression of its CD19 antigen was detected by flow cytometry, shown below.
Figure 1. Flow cytometric detection of CD19 antigen expression in CD19-293T stably transfected cell line.
2. CAR-T cell phenotype analysis and CAR positive rate test results
Monitoring the activated and expanded PBMC, the results showed that high-purity T cells were successfully obtained and which accounted for more than 90% of the cells screened. The subtype analysis results of T cells showed that the proportions of CD4+ and CD8+ cells were 35.93% and 49.45%. Furthermore, FMC63 CAR-T cells were constructed by lentiviral transduction, and FMC63 CAR-T cells were labeled with FMC63 scFv antibodies. The positive rate of FMC63 CAR was 16.99%, indicating that FMC63 CAR-T cells were successfully constructed.
Figure 2. T cell phenotype analysis and FMC63 CAR-T cell positive rate detection. (A) The proportion of T cells on the tenth day of PBMC activation and expansion and the results of the phenotypic analysis. (B) FMC63 CAR-T cell CAR positive rate test result.
In vivo evaluation models
Services：
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Type
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Model
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Service |
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Animal model |
Immunodeficient Mice
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BALB/c nude mice, NOD scid mice, C-NKG mice
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Humanized mice with immune system
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Hu-PBMC mice, Hu-HSC mic
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Cell Line-derived Tumor Xenograft (CDX) models
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Subcutaneous solid tumor model, In situ solid tumor model, Hematoma model
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Homologous tumor mouse model
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Lung cancer, colorectal cancer, breast cancer, melanoma, and other homologous tumor models
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Cell Line-derived Tumor Xenograft (CDX) models
Humanized Immune System (HIS) Models
Case Study
1. Detection of C-NKG Mouse-derived Immune Components in Severe Immunodeficiency Mice
Figure 3. The B, T, and NK cells in the peripheral blood of C-NKG mice are severely lacking, making them a severely immunodeficient mouse.
The peripheral blood of BALB/c and C-NKG mice was sampled and used to perform representative flow cytometric immunophenotyping and statistical comparison on the composition of T, B and NK cells. The results showed that, compared with BALB/c mice, B cells (CD3-CD19+), T cells (CD3+ CD19-), helper T cells (CD3+ CD4+ CD8-) and cytotoxicity in the peripheral blood of C-NKG mice T cells (CD3+ CD4- CD8+) and NK cells (CD335+ CD3-) are almost completely absent.
2. Selected Cell line Derived Xenografts (CDX) Model
Figure 4. Formation experiment of A549 Human non-small cell lung cancer (NSCLC) cell subcutaneous tumor.
Figure 5. Photos of nude mice inoculated with A459 cells 6 weeks after inoculation.
Inquiries and Quote Requests
Request a quote now. Alternatively, you can always email service-apac@cyagen.com or call 86 20-31601779 to inquire about our services or obtain a quote for your project.
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