Humanized Brain Tumor Models

Traditional animal models often fail to replicate the complex human tumor microenvironment and immune interactions that are critical for studying tumor behavior and treatment responses. Humanized mouse models address these limitations by integrating human immune cells and tumor tissues, allowing for a more accurate representation of human diseases. Alfa Cytology offers bespoke humanized model development to meet the specific needs of our clients.

Introduction to Humanized Brain Tumor Models

The development of humanized brain tumor models involves the engraftment of human hematopoietic stem cells (HSCs) into immunodeficient mice, such as DRAG (NOD.Rag1KO.IL2Rγ cKO) mice. These models can be used to study the pathological features of GBM and the interactions between tumor cells and the human immune system.

Fig.1 Construction of Humanized Brain Tumor Models. (Srivastava R., et al., 2023)

For instance, the incorporation of human immune cells allows researchers to investigate how these cells respond to various therapies, including immune checkpoint inhibitors like anti-PD-1 antibodies. The results from such studies not only enhance our understanding of tumor biology but also pave the way for the development of novel therapeutic strategies.

Humanization of Rodents

The humanization of rodents, including mice and rats, can be achieved through two primary methods:

Cellular Humanization: This method involves the xenograft transplantation of human cells or tissues to supply human leukocytes, such as CD34+ hematopoietic stem cells, peripheral blood mononuclear cells (PBMCs), or purified leukocyte subpopulations.
Genetic Humanization: This approach entails replacing mouse genes with their human counterparts, which can either create relevant targets for human-specific therapies or provide essential support for the generation, maintenance, and functionality of the transplanted human cells.

Our Services

At Alfa Cytology, we specialize in providing high-quality preclinical models for brain tumor research. Our humanized brain tumor models are designed to facilitate comprehensive studies of GBM and other brain cancers, enabling researchers to assess therapeutic efficacy and mechanisms of action in a relevant biological context. Our comprehensive services encompass the entire spectrum of preclinical research, from model generation to efficacy testing of therapeutic agents.

Immune System Reconstruction Models Development

Alfa Cytology specializes in the development of advanced immune system reconstruction models to enhance cancer research and immunotherapy studies.

Human Hematopoietic Stem-Cell (HHSC) Based Models Development
Human PBMC Reconstituted Models Development

Genetic Humanization Model Development

Alfa Cytology employs employ advanced techniques to replace mouse genes with their human counterparts, creating relevant targets for human-specific therapies.

Whole Gene Humanization - Swapping in Human Sequence for Animal Gene
Point Mutation Humanization - Inserting Clinical Variation into Animal's Gene

Case Study - Humanized DRAG Mouse Model

Model Introduction

The humanized DRAG mouse model provides a sophisticated preclinical platform for studying glioblastoma immunotherapy in the context of a fully functional human immune system. Generated by engrafting human CD34+ HLA-DR4+ hematopoietic stem cells into immunodeficient DRAG mice, the model enables development of a complete human immune cell repertoire. Intracranial implantation of patient-derived GBM tumor sphere cells faithfully recapitulates the histopathological features and tumor immune microenvironment of human GBM, enabling preclinical evaluation of immune checkpoint inhibitors such as anti-PD-1 antibodies.

Model Information

Model: Humanized DRAG Mouse Model
Animal: DRAG Mice (NOD.Rag1KO.IL2RγcKO, HLA-DR4 transgenic)
Weight: 20–30 g

Cancer Type: Glioblastoma (GBM)
Age: 8–12 Weeks

Model Construction

The model is established through a sequential two-phase construction protocol.

Human Immune System Reconstitution Phase: DRAG mice receive 250 cGy whole-body irradiation, followed by intravenous injection of 2 × 10⁵ human CD34+ HLA-DR4+ hematopoietic stem cells. Engraftment is monitored for 10–12 weeks to achieve stable human immune cell reconstitution.
Orthotopic Tumor Implantation Phase: Patient-derived GBM tumorsphere cells (2 × 10⁵ cells) are stereotactically implanted intracranially. Tumor growth is monitored using bioluminescence imaging and MRI.

Fig. 2 Workflow of humanized DRAG mouse model establishment. (Source: Alfa Cytology)

Model Data

Human Immune Reconstitution: Human CD45+ cells constitute 11.6% of PBMCs and 13.2% of splenocytes. Reconstituted immune cells include T cells (40%), B cells (54.3%), monocytes (19.5%), and NK cells (14.2%). Regulatory T cells are detected in both PBMCs and spleen.
Tumor Histopathology: Tumors recapitulate human GBM features, including pseudopalisading necrosis, microvascular proliferation, and expression of GFAP, Olig2, Nestin, and SOX2.
Therapeutic Efficacy: Anti-PD-1 antibody treatment (10 mg/kg) significantly suppresses tumor growth, prolongs survival, and decreases immunosuppressive cell populations, demonstrating the model's utility for evaluating immune checkpoint inhibitors.

Fig. 3 Anti-PD-1 response in humanized DRAG mouse GBM model. Data are presented as mean ± standard error (SEM). (Source: Alfa Cytology)

Contact Us

To learn more about our humanized brain tumor models and how they can enhance your research, please contact us at Alfa Cytology. Our team of experts is ready to assist you with any inquiries and to discuss how our services can be tailored to meet your specific research needs.

Reference

Srivastava R., et al. (2023) Development of a human glioblastoma model using humanized DRAG mice for immunotherapy[J]. Antibody Therapeutics. 6(4): 253-264.

All of our services and products are intended for preclinical research use only and cannot be used to diagnose, treat or manage patients.