A novel GFP nude rat model to investigate tumor-stroma interactions

Cancer Cell International

Jian Wang²  Xingang Li²  Per Øyvind Enger³  Lina Leiss⁴  Huaiyang Zhu¹  Narve Brekkå¹  Oleg Tsinkalovsky¹  Bin Huang²  Ning Yang² 

[1] Department of Biomedicine, University of Bergen, Bergen, N-5009, Norway;Brain Science Research Institute, Shandong University, Jinan, China;Department of Neurosurgery, Haukeland University Hospital, Bergen, Norway;Neuro Clinic, Haukeland University Hospital, Bergen, Norway

关键词: Tumor biology;    Breast cancer;    Glioblastoma;    Tumor-stroma interaction;    Xenograft;   

Others  :  1171063
DOI  :  10.1186/s12935-014-0146-0

 received in 2014-09-10, accepted in 2014-12-11,  发布年份 2014

【 摘 要 】

Backgroud

A key strategy for the study of the tumor microenvironment is to implant human tumor cells in an immunodeficient rodent strain ubiquitously expressing a fluorescent marker. Here, a novel nude rat expressing a green fluorescent protein (GFP) transgene was established and engrafted with primary human tumor tissue in order to separate tumor from stromal cell populations for subsequent molecular analysis.

Methods

SD-TG (GFP) 2BalRrrc transgenic rats were crossed with HsdHan™: rnu/rnu Rowett nude rats to develop a GFP expressing immunocompromised rat. PCR and flow cytometry were used to follow the GFP genotype and phenotype in newborns. After three to four generations, animals were implanted with 4 T1 dsRed murine breast cancer cells or primary human glioblastoma (GBM) biopsies to generate xenografts for subsequent separation by fluorescence-activated cell sorting (FACS).

Results

Fluorecence microscopy and reverse transcription-PCR demonstrated that GFP, under the control of the human ubiquitin C promoter, was stably maintained and expressed in diverse organs over several generations. Immunophenotyping of blood samples by flow cytometry confirmed that the immunodeficient features of the parental rat strain, HsdHan™: rnu/rnu, were retained in the GFP nude rat. Both the murine cell line and human GBM biopsies engrafted, and stromal cell populations were isolated from dissociated xenografts by FACS to > 95% purity.

Conclusions

A GFP transgene was stably introduced into a nude rat background in which human and murine cancer cells successfully engrafted. This animal strain provides a novel in vivo system for detailed cellular and molecular characterization of tumor-stroma interactions.

【 授权许可】

   
2014 Yang et al.; licensee BioMed Central.

【 图 表 】

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