C57BL/6-Cd3etm1(CD3E)Bcgen Cd3dtm1(CD3D)Bcgen Cd3gtm1(CD3G)Bcgen Epcamtm1(EPCAM)Bcgen/Bcgen • 113797
Product name | B-hCD3EDG/hEPCAM mice |
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Catalog number | 113797 |
Strain name | C57BL/6-Cd3etm1(CD3E)Bcgen Cd3dtm1(CD3D)Bcgen Cd3gtm1(CD3G)Bcgen Epcamtm1(EPCAM)Bcgen/Bcgen |
Strain background | C57BL/6 |
NCBI gene ID | 916,915,917,4072 (Human) |
Aliases | IMD18, T3E, TCRE; CD3-DELTA, IMD19, T3D; CD3-GAMMA, IMD17, T3G; DIAR5, EGP-2, EGP314, EGP40, ESA, HNPCC8, KS1/4, KSA, M4S1, MIC18, MK-1, TACSTD1, TROP1 |
Strain specific analysis of EPCAM mRNA expression in wild-type C57BL/6 mice and B-hCD3EDG/hEPCAM mice by RT-PCR. Small intestine RNA were isolated from wild-type C57BL/6 mice (female, 6-week-old, n=1) and homozygous B-hCD3EDG/hEPCAM mice (female, 6-week-old, n=1), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mouse or human EPCAM primers. Mouse Epcam mRNA was only detectable in wild-type mice (+/+). Human EPCAM mRNA was exclusively detectable in homozygous B-hCD3EDG/hEPCAM mice (H/H) but not in wild-type mice (+/+).
CD3E expression analysis in wild-type C57BL/6 mice and homozygous B-hCD3EDG/hEPCAM mice by flow cytometry. Spleen and blood T cells were collected from wild-type C57BL/6 mice (female, 6-week-old, n=1) and homozygous B-hCD3EDG/hEPCAM mice (female, 6-week-old, n=1). Protein expression was analyzed with anti-human CD3E antibody (BD Horizon™, 562426) and anti-mouse CD3E antibody (Biolegend, 100312) by flow cytometry. Human CD3E was exclusively detectable in homozygous B-hCD3EDG/hEPCAM mice, but not in wild-type C57BL/6 mice.
Immunohistochemical (IHC) analysis of EPCAM expression in B-hCD3EDG/hEPCAM mice. The kidney, heart, lung, liver, skin, small intestine, large intestine and spleen were collected from wild-type C57BL/6 mice and B-hCD3EDG/hEPCAM mice (female, 6-week-old), analyzed by IHC with anti-hEPCAM (abcam, ab213501). Mouse EPCAM was detectable in C57BL/6 mice. The arrow indicates tissue cells with positive EPCAM staining (brown). “+” indicates that the tissue is positive, and “-” indicates that the tissue is negative.
Immunohistochemical (IHC) analysis of EPCAM expression in B-hCD3EDG/hEPCAM mice. The kidney, heart, lung, liver, skin, small intestine, large intestine and spleen were collected from wild-type C57BL/6 mice and B-hCD3EDG/hEPCAM mice (female, 6-week-old), analyzed by IHC with anti-hEPCAM (abcam, ab223582). Human EPCAM was detectable in B-hCD3EDG/hEPCAM mice. The arrow indicates tissue cells with positive EPCAM staining (brown). “+” indicates that the tissue is positive, and “-” indicates that the tissue is negative. This antibody exhibited strong non-specific binding in the small intestine and liver, with no obvious membrane staining signals observed in the bronchial epithelial cells of the lung.
Frequency of leukocyte subpopulations in spleen by flow cytometry. Splenocytes were isolated from wild-type C57BL/6 mice (female, n=3, 6-week-old) and homozygous B-hCD3EDG/hEPCAM mice (female, n=3, 8-week-old). A. Flow cytometry analysis of the splenocytes was performed to assess the frequency of leukocyte subpopulations. B. Frequency of T cell subpopulations. Percentages of T cells, B cells, NK cells, dendritic cells, neutrophils, monocytes, macrophages, CD4+ T cells, CD8+ T cells and Tregs in B-hCD3EDG/hEPCAM mice were similar to those in C57BL/6 mice, demonstrating that humanization does not change the frequency or distribution of these cell types in spleen. The frequency of leukocyte subpopulations in blood and lymph node of B-hCD3EDG/hEPCAM mice were also comparable to wild-type C57BL/6 mice (Data not shown).