Orthotopic model of SHG-44 in the enhanced green fluorescent protein nude mouse.

Naturally fluorescent proteins have been widely used in biological research. In this study, we found that the simple and effective way to obtain enhanced green fluorescent protein (EGFP) nude mice is to cross transgenic EGFP C57BL/6J mice with nude (nu/nu) mice. EGFP expression is identified by tail genotyping. Establishment of the orthotopic EGFP nude mouse model used surgical orthotopic implantation. The morphology and human glioma cell markers, such as glial fibrillary acidic protein (GFAP) and S-100, remain unchanged in this mouse model. The tumor blood vessels obtained from the orthotopic model show brilliant EGFP fluorescence as observed by fluorescence microscopy. These findings suggested that this is an ideal mouse model with which to study interaction among host, tumor, and tumor microenvironment; the findings also suggested that the host (EGFP nude mouse) was involved in tumor angiogenesis.

Anti-human CD133 monoclonal antibody that could inhibit the proliferation of colorectal cancer cells.

CD133, a five-transmembrane molecule, has been found on many types of cancers and determined to be a cancer stem cell biomarker. In this study a functional anti-human CD133 MAb 6B6 was obtained, and the specificity of this MAb was verified by flow cytometry. This MAb effectively recognized the CD133 molecule expressed on a series of malignant cell lines. Immunohistochemistry staining showed the CD133 was expressed on colorectal tumor tissue. Furthermore, we demonstrated that MAb 6B6 could inhibit the proliferation of Caco-2 cells that were derived from a human colorectal carcinoma. This functional anti-human CD133 MAb provides a valuable tool for further study of biological functions of cancer stem cell that expressed CD133.

[Transplantation of human glioma stem cells in nude mice with green fluorescent protein expression].

OBJECTIVES: To investigate the possibility of transplantation of human glioma stem cells (HGSCs) in nude mice stably expressing green fluorescent protein (GFP) so as to clearly identify the incubated HGSCs from the host tissues. METHODS: Transgenic C57BL/6J mice expressing GFP was crossed with nude mice of the line NC, then hairless male nude mice expressing GFP were crossed with hairy female pubescent mice to obtain nude mice with GFP expression the expression of GFP in the skin and organs of these nude mice were evaluated by naked eyes, and immunohistochemical and immunofluorescence assays. HGSCs were transplanted orthotopically into the caudate nuclei of nude mice expressing GFP. Immunohistochemistry was used to observe the transplanted tumor. RESULTS: The structures rich in adipose tissue of the 8th generation nude mice were dark green and the other organs were light green. However, green fluorescence was emitted from all tissues under fluorescence microscopy. Confocal fluorescence microscopy showed that the tumor cells were stained red, distinguished from the host cells distinctly in the brains bearing tumor transplanted orthotopically. CONCLUSION: Nude mice expressing GFP can be obtained by crossing the transgenic mice bearing naive immunity with nude mice. Orthotopic transplantation of HGSCs may be used in the investigation of tumor tissue reconstitution because of the easy identification between the transplantation tumor and host tissue.

[Immune response and immunopathology in inducible costimulatory molecule (ICOS) transgenic mice infected with Schistosoma japonicum].

OBJECTIVE: To establish the ICOS transgenic mice schistosomiasis japonica model and to observe the immune response and immunopathology of the model. METHODS: The transgenic mice were infected with Schistosoma japonicum. Spleen cells and sera of mice were harvested at week 4, 6, and 8 after infection. The cytokines IFN-gamma and IL-4 were measured in culture supernatans by ELISA. The serum IgG, IgGI and IgG2a were measured by ELISA at different period of infection. Liver tissue sections were prepared with HE staining. Liver granuloma formation was observed under microscope. RESULTS: The expression level of IFN-y showed no significant difference between ICOS transgenic mice and control, while that of IL-4 in ICOS transgenic mice was significantly up-regulated to 20.81+/-1.95 and 25.31+/-3.37 pg/ml at week 6 and 8 respectively (P<0.01). The serum IgG and IgGl in ICOS transgenic mice were also significantly higher than those in control. Th2 differentiation index and lgC1/IgG2a were used to evaluate the immune regulation balance of Thl/Th2, and results showed that Th2 response in ICOS transgenic mice was significantly stronger than that of the control. The egg granuloma response in ICOS transgenic mice was also significantly stronger than that in control (P<0.01). The rate of egg granuloma enlargement was 24.48% and 26.37% at week 6 and 8 respectively. CONCLUSION: The findings suggest that there is stronger Th2 type response in ICOS transgenic mice infected with Schistosoma japonicum and ICOS may play an important role in the egg granuloma formation of Schistosoma japonicum.

[The study of sperm cryopreservation in kunmin (KM) mouse].

Spermatozoa of 10, 15 and 20-wk-old KM mice were frozen and stored at--196 degrees C. After thawing, intact superovulation oocytes were inseminated in vitro with relative high motility frozen/thawed mouse spermatozoa. Some 2-cell embryos obtained by in vitro fertilization were transferred to pseudopregnant recipients and 47%-56% of them developed into live-born offspring. Some 2-cell embryos were incubated in vitro and 57% of them progressed to the blastocyst stage. Others were cryopreserved by simple vitrification procedure, and most of them (about 80%) were morphologically normal after warming, and 50% of the cryopreserved embryos transferred to recipients had developed into live pups. These results indicated that there was no significant difference when compared with embryos obtained from oocytes inseminated with fresh sperm.