General information

Development

CSF2, also known as GM-CSF (Granulocyte-Macrophage Colony-Stimulating Factor), is a hematopoietic growth factor that stimulates the development of neutrophils and macrophages, and promotes the proliferation and development of early erythroid and eosinophil progenitor cells. To this end, Vitastar has used a transgenic approach to amplify the fragment upstream of mouse CSF1's ATG by 1.2Kb (containing the promoter), and linked it to the cDNA of human CSF2, inserting it into the pCDNA3.1 vector to construct the pCDNA3.1-Csf1p-hCSF2 expression vector (as shown in the figure below). The expression vector fragment was isolated by enzymatic digestion and injected into the pronucleus of NPG mice. Positive mice with appropriate expression levels were selected from the offspring, and in isolators, bred in a manner of inbreeding to establish a commercialized hCSF2 NPG mouse strain.

The hCSF2 NPG mice express an appropriate amount of human CSF2 protein, and together with NPG mice expressing human M-CSF and IL3 cell factors, after the transplantation of human CD34+ hematopoietic stem cells for 6-8 weeks, form stable and widespread myeloid and lymphoid cell differentiation in peripheral blood, bone marrow, thymus, spleen, and non-lymphatic tissues including lungs and liver. In the blood and tissues, differentiation of granulocytes (basophils, neutrophils, and eosinophils), antigen-presenting cell differentiation (dendritic cells and macrophages), and regulatory T cell populations can be detected. This mouse model has more precise applications in humanization, cancer treatment, human allergic response models, infectious diseases, immunology, and regenerative medicine research.

Fig1. Schematic diagram of NPG-hCSF2 construction

Phenotype

1. Analysis of immune cell reconstitution of transplanted umbilical cord blood CD34⁺ cells

Tab1. hCSF2-NPG mice were analyzed for peripheral blood flow cytometry 3 weeks after HSC transplantation

NPG-hCSF2 Mice Applications

1. Humanization and cancer therapy

2. Human allergic reaction modeling, immune and hematopoietic transplantation reconstruction modeling

3. Regenerative medicine

4. Infectious diseases

Reference

1. Wunderlich M, Chou FS, Link KA, et al. AML xenograft efficiency is significantly improved in NOD/SCID-IL2RG mice constitutively expressing human SCF, GM-CSF and IL-3. Leukemia. 2010, 24(10):1785-1788.

2. Ito R, Takahashi T, Katano I, et al. Establishment of a human allergy model using human IL-3/GM-CSF-transgenic NOG mice. J Immunol. 2013, 191(6):2890-2899.

3. Rongvaux A, Willinger T, Martinek J, et al. 2014. Development and function of human innate immune cells in a humanized mouse model. Nat Biotechnol 32(4):364-372.