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RESEARCH MODELS

HuCD34-NCG Mouse Model

Product Overview A mouse model expressing human cells (humanized) is a beneficial tool to study diseases and develop therapeutics.

Highly immunodeficient animals are required to support the engraftment of human cells. The triple-immunodeficient NCG

(NOD-Prkdcem26Cd52Il2rgem26Cd22/NjuCrl) mouse is ideal for developing these valuable in vivo humanized models because the

strain lacks functional murine T cells, B cells, and natural killer (NK) cells, in addition to showing reduced macrophage and

dendritic cell function. The immunodeficient NCG mouse is an effective research platform for humanization.

HuCD34-NCG Mouse ModelNCG mice are humanized by adoptive transfer using human umbilical cord blood-derived CD34+ stem cells from a qualified

source, following myeloablation treatment. Animals are subsequently housed for 12-15 weeks according to Charles River

immunodeficient animal housing protocols. During this period, CD34+ stem cells develop into human immune cells that

engraft within the immunodeficient NCG mice. The presence of human immune cells are verified in the peripheral blood as

early as 12-15 weeks post-engraftment as verified by multi-color flow cytometry. These immune cells are detectable in mouse

tissues, including bone marrow, spleen, liver, and lung, and have been shown to infiltrate implanted tumors. The HuCD34-NCG

is an ideal in vivo platform to evaluate the effectiveness of compounds that modulate the human immune system. The lack of,

or late onset of, graft-versus-host-disease (GvHD) in humanized mice make them ideal for long-term studies.

Humanized Mice

Myeloablation by whole body γ irradiation of 4-6 week old NCG mice

I.V. injection of human CD34+ stem cells

Ideal for research in oncology, immunology, infectious disease,

regenerative medicine, and human organ transplantation

12–15 weeks

Development of a human

immune system

SummaryIntroducing the new study-

ready HuCD34-NCG mouse

model, with a human-like

immune system.

Advantages

• Convenient: HuCD34-NCG mice are study-ready, expressing human immune cells

• Consistent: Every mouse is pre-tested for humanization

• Stable: Engraftment of hematopoietic cells provides a research platform for extended studies

• License-free: No additional licensing fees or royalty payments

Engraftment Data and Model Performance

Engraftment rates

92 124 180 236

100

90

80

70

Days Post Injection

% h

uman

izat

ion

100 101 102 103 104 105 106

106

105

104

103

102

101

100

hCD45+

mC

D45

+

mCD45+4.26

hCD45+41.4

Figure 1: Human immune cell engraftment. Whole blood collected via submandibular bleed was analyzed at Days 92, 124, 180, and 236 (Weeks 13, 18, 26, and 34) post-injection using a 10-color flow cytometry panel. The presence of human lymphoid cells is calculated as percentage of human CD45+ cells of total lymphocytes detected.

Analysis of human immune cell populations

Figure 2: Immunophenotyping of immune cells in HuCD34-NCG mice. Whole blood collected via submandibular bleed and analyzed using a 10-color flow cytometry panel at Days 92, 124, 180, and 236 (Weeks 13, 18, 26, and 34) post-injection. Levels of peripheral total T cells (CD3+) and T cell subsets (CD4+ and CD8+), B cells (CD19+), and NK cells (CD3-CD16+CD56+) were detected throughout the study (data not shown for B cells and NK cells). Relative percentages are given as a proportion of the respective parent population.

Example of tumor study using HuCD34-NCG mouse model

Figure 3: A549 tumor-inoculated HuCD34-NCG animals respond to anti-hPD1 dosing regimen. A549 (human lung adenocarcinoma cell line) tumors were implanted subcutaneously on the left flank of HuCD34-NCG mice. The animals were treated with a regimen of anti-hPD1 therapy and tumor volumes were measured twice weekly. Spleen and tumor samples of HuCD34-NCG animals were collected at study termination for immunophenotyping by flow cytometry and T cell infiltration evaluated. Frequencies of analyzed subsets presented as proportion of their respective parent population in the gating hierarchy: CD4+ and CD8+ T cells as percentage of CD3+ T cells and Treg as percentage of CD4+ T cells.

For additional information on our humanized model portfolio, please visit www.criver.com/humanizedmodels.

For more information on this model, please contact Technical Support at TAD@crl.com.

To order the HuCD34-NCG, please contact our Customer Service Department at 1-800-522-7287

or via email orders@crl.com.

Research Applications

• Oncology

• Immunology

• Hematopoiesis

• Stem cell research

• Infectious disease

• Regenerative medicine

• Transplantation research

• Hematological disease

Applications

• Tumor-bearing NCG – Mice expressing a full complement of human immune cells that are engrafted with cell-derived xenografts (CDX) and patient-derived xenografts (PDX) are necessary for immuno-oncology therapeutics.

• Infectious disease research – Humanized mice are powerful tools to study immune responses to foreign agents, including HIV and compounds used to treat infectious diseases.

• Immunology – Humanized mouse models can be used to study cellular development (hematopoiesis) and immune cell function.

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