A novel GFP nude rat model to investigate tumor-stroma interactions
Yang, Ning; Huang, Bin; Tsinkalovsky, Oleg; Brekkå, Narve; Zhu, Huaiyang; Leiss, Lina; Enger, Per Øyvind; Li, Xingang; Wang, Jian
Peer reviewed, Journal article
Published version
Permanent lenke
https://hdl.handle.net/1956/9396Utgivelsesdato
2014-12-21Metadata
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Originalversjon
https://doi.org/10.1186/s12935-014-0146-0Sammendrag
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.
Utgiver
BioMed CentralTidsskrift
Cancer Cell InternationalOpphavsrett
Copyright 2014 Yang et al.; licensee BioMed CentralNing Yang et al.; licensee BioMed Central Ltd.