Novel enhanced GFP-positive congenic inbred strain establishment and application of tumor-bearing nude mouse model

Qing Lan; Yanming Chen; Chungang Dai; Shenggang Li; Xifeng Fei; Jun Dong; Yanhua Shen; Xingliang Dai; Zhaohui Lu; Bing Liu; Qilong Wang; Haiyang Wang; Zhengyu Zhou; Xiaoyan Ji; Zhimin Wang; Qiang Huang

doi:10.1111/cas.14545

Novel enhanced GFP-positive congenic inbred strain establishment and application of tumor-bearing nude mouse model

Cancer Sci. 2020 Oct;111(10):3626-3638. doi: 10.1111/cas.14545. Epub 2020 Aug 11.

Authors

Qing Lan¹, Yanming Chen¹, Chungang Dai¹, Shenggang Li¹, Xifeng Fei², Jun Dong¹, Yanhua Shen³, Xingliang Dai⁴, Zhaohui Lu¹, Bing Liu¹, Qilong Wang¹, Haiyang Wang¹, Zhengyu Zhou³, Xiaoyan Ji⁵, Zhimin Wang², Qiang Huang¹

Affiliations

¹ Department of Neurosurgery, The Second Affiliated Hospital of Soochow University, Suzhou, China.
² Department of Neurosurgery, Suzhou Kowloon Hospital of Shanghai Jiaotong University School of Medicine, Suzhou, China.
³ Laboratory Animal Center, Soochow University, Suzhou, China.
⁴ Department of Neurosurgery, The First Affiliated Hospital of Anhui Medical University, Hefei, China.
⁵ Department of Ophthalmology, The Second Affiliated Hospital of Soochow University, Suzhou, China.

Abstract

Transgenic GFP gene mice are widely used. Given the unique advantages of immunodeficient animals in the field of oncology research, we aim to establish a nude mouse inbred strain that stably expresses enhanced GFP (EGFP) for use in transplanted tumor microenvironment (TME) research. Female C57BL/6-Tg(CAG-EGFP) mice were backcrossed with male BALB/c nude mice for 11 generations. The genotype and phenotype of novel inbred strain Foxn1^nu .B6-Tg(CAG-EGFP) were identified by biochemical loci detection, skin transplantation and flow cytometry. PCR and fluorescence spectrophotometry were performed to evaluate the relative expression of EGFP in different parts of the brain. Red fluorescence protein (RFP) gene was stably transfected into human glioma stem cells (GSC), SU3, which were then transplanted intracerebrally or ectopically into Foxn1^nu .B6-Tg(CAG-EGFP) mice. Cell co-expression of EGFP and RFP in transplanted tissues was further analyzed with the Live Cell Imaging System (Cell'R, Olympus) and FISH. The inbred strain Foxn1^nu .B6-Tg(CAG-EGFP) shows different levels of EGFP expression in brain tissue. The hematological and immune cells of the inbred strain mice were close to those of nude mice. EGFP was stably expressed in multiple sites of Foxn1^nu .B6-Tg(CAG-EGFP) mice, including brain tissue. With the dual-fluorescence tracing transplanted tumor model, we found that SU3 induced host cell malignant transformation in TME, and tumor/host cell fusion. In conclusion, EGFP is differentially and widely expressed in brain tissue of Foxn1^nu .B6-Tg(CAG-EGFP), which is an ideal model for TME investigation. With Foxn1^nu .B6-Tg(CAG-EGFP) mice, our research demonstrated that host cell malignant transformation and tumor/host cell fusion play an important role in tumor progression.

Keywords: congenic inbred green fluorescent nude mice; dual-fluorescence tracing model; glioma stem cells; malignant transformation; tumor microenvironment.

MeSH terms

Animals
Brain / physiology
Cell Fusion / methods
Cell Line, Tumor
Cell Transformation, Neoplastic / genetics
Disease Models, Animal
Female
Glioma / genetics*
Green Fluorescent Proteins / genetics*
Luminescent Proteins / genetics
Male
Mice
Mice, Inbred BALB C
Mice, Inbred C57BL
Mice, Nude
Mice, Transgenic
Neoplastic Stem Cells / pathology
Red Fluorescent Protein
Transfection / methods
Transplantation, Heterologous / methods
Tumor Microenvironment / genetics

Substances

Luminescent Proteins
Green Fluorescent Proteins

Abstract

MeSH terms

Substances

Grants and funding