A short-term in vivo model for giant cell tumor of bone

Abstract

BackgroundBecause of the lack of suitable in vivo models of giant cell tumor of bone (GCT), little is known about its underlying fundamental pro-tumoral events, such as tumor growth, invasion, angiogenesis and metastasis. There is no existing cell line that contains all the cell and tissue tumor components of GCT and thus in vitro testing of anti-tumor agents on GCT is not possible. In this study we have characterized a new method of growing a GCT tumor on a chick chorio-allantoic membrane (CAM) for this purpose.MethodsFresh tumor tissue was obtained from 10 patients and homogenized. The suspension was grafted onto the CAM at day 10 of development. The growth process was monitored by daily observation and photo documentation using in vivo biomicroscopy. After 6 days, samples were fixed and further analyzed using standard histology (hematoxylin and eosin stains), Ki67 staining and fluorescence in situ hybridization (FISH).ResultsThe suspension of all 10 patients formed solid tumors when grafted on the CAM. In vivo microscopy and standard histology revealed a rich vascularization of the tumors. The tumors were composed of the typical components of GCT, including (CD51+/CD68+) multinucleated giant cells whichwere generally less numerous and contained fewer nuclei than in the original tumors. Ki67 staining revealed a very low proliferation rate. The FISH demonstrated that the tumors were composed of human cells interspersed with chick-derived capillaries.ConclusionsA reliable protocol for grafting of human GCT onto the chick chorio-allantoic membrane is established. This is the first in vivo model for giant cell tumors of bone which opens new perspectives to study this disease and to test new therapeutical agents.

Bortezomib Inhibits Giant Cell Tumor of Bone through Induction of Cell Apoptosis and Inhibition of Osteoclast Recruitment, Giant Cell Formation, and Bone Resorption

“…Stromal cells injected subcutaneously into immunocompromised mice do not produce giant cells (44)(45)(46). Moreover, tumor tissues grown on chick chorioallantoic membranes do not appear to recruit chicken monocytes to synthesize new giant cells and are unsuitable for drug administration, despite increased vascularization from the membrane, but only survive for 10 days (5,31). In our study, we used patient-derived tumor cells isolated from GCTB to establish an orthotopic murine model by intratibial injection.…”

New ex-ovo colorectal-cancer models from different SdFFF-sorted tumor-initiating cells

“…Advantages are that the CAM model lacks immunological and inflammatory cells, making it an attractive system to avoid effects of the tumor microenvironment interfering with tumor development [6]. This has been used to provide highly reproducible models for glioblastoma [7,8], bone and osteosarcoma [5,9], pancreatic adenocarcinoma [10], ovarian cancer [11], head and neck squamous carcinoma [12], and more recently renal carcinoma [13], as well as CRC [14]. In the last case, the development of CRC and therapeutic escape may be caused by the presence of tumor-initiating cells (TICs) [3,15], which are responsible for driving growth and treatment resistance [16,17].…”

“…Spontaneous tumor initiation from human tumor cells grafted Carole Mélin, Aurélie Perraud, Serge Battu and Muriel Mathonnet contributed equally to this work. on the CAM has provided valuable information in a short time regarding such crucial processes as tumor cell proliferation and invasion corresponding to the first steps of carcinogenesis [5]. Advantages are that the CAM model lacks immunological and inflammatory cells, making it an attractive system to avoid effects of the tumor microenvironment interfering with tumor development [6].…”

Ethical euthanasia and short-term anesthesia of the chick embryo

“…On the 5 th day of embryonic development, the CAM attaches to the inner eggshell membrane (Romanoff, 1960) and opening of the egg without rupturing this structure is no longer possible. The procedure was performed as described by Kunzi-Rapp et al (2001) and Balke et al (2011) with a few modifications. Briefly, the eggs were washed with warm (40 ± 5°C) 70% ethanol and placed on a six well plate in horizontal position.…”