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

Abstract

BackgroudA 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.MethodsSD-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).ResultsFluorecence 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.ConclusionsA 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.

Evaluation of complete cytoreductive surgery and two intraperitoneal chemotherapy techniques in pseudomyxoma peritonei

“…The animals underwent the same degree of CRS, and the impact of variations in CC-score could not be evaluated. Tumor recurrence in all animals during follow-up should be viewed in light of several unfavorable factors; the use of immunodeficient animals which was necessary in order to allow growth of human tumor xenograft [288]; an aggressive tumor model with mucinous tumor with signet ring cells [289]; tumor distribution on critical sites allowed CRS with CC-1. A study from a major institution, in which patients with PMP were retrospectively stratified according to the PSOGI classification, have also demonstrated extremely poor survival after CRS-HIPEC in patients with high-grade PMP with signet ring cells, i.e.…”

Organism-Level Tumor Models in Zebrafish Danio rerio

“…Obviously, any analysis of the interaction between a tumor and stromal tissue is impossible without the use of organism-level models. The most common, currently used systems are based on immunodeficient rodent lines, which provide a combination of an organism-level model, fluorescent microscopy, FACS analysis of cell subpopulations, and transcription analysis [ 48 ]. However, the D. rerio model is gaining in popularity, especially where high-resolution microscopy is warranted [ 49 , 50 ].…”