3D chick embryo chorioallantoic membrane model as an in vivo model to study morphological and histopathological features of feline fibrosarcomas

Abstract

BackgroundThe chick embryo chorioallantoic membrane (CAM) model is well described in human medicine as a cost-effective, easy to perform preclinical oncological model for observing pro- and antiangiogenic response, tumor biology and metastasis. The main objective of this article was to present the modification of the CAM assay in order to evaluate tumor growth from two feline fibrosarcoma cell lines (FFS1, FFS3) and describe their morphological and histopathological features.ResultsThe authors described morphological and histopathological features of two feline fibrosarcoma cell lines (FFS1 and FFS3) grown on the CAM. Tumors from the FFS1 cell line showed high malignancy (grade III), while tumors from the FFS3 cell line were grade II. Proliferation markers (Ki-67 and PCNA) were determined and the positive correlation between PCNA and tumor grade (r = 0.8247; p < 0.001) was demonstrated, as opposed to Ki-67.ConclusionsThe results obtained indicate that PCNA may be helpful to evaluate the tumor grade, better than Ki-67, for feline fibrosarcomas. However, further investigations of proliferation marker, in bigger number of feline spontaneous fibrosarcomas and feline fibrosarcomas grown on the CAM from different cell lines, are needed to confirm these observations.

Distribution of Glutathione-Stabilized Gold Nanoparticles in Feline Fibrosarcomas and Their Role as a Drug Delivery System for Doxorubicin—Preclinical Studies in a Murine Model

“…FISS growth from the FFS1 cell line in a murine model were grade III with very fast growth and tumor enlargement ( Figure 12 ), which is in agreement with the previous studies on the chick embryo chorioallantoic membrane model [ 20 ]. Some of the tumors were attached to the vertebrae or scapula of the mice, indicating the highly invasive nature of FISS, which is in agreement with the clinical observations in cats [ 1 , 21 ].…”

Proteomic Differences in Feline Fibrosarcomas Grown Using Doxorubicin-Sensitive and -Resistant Cell Lines in the Chick Embryo Model

“…The advantages of using the chicken embryo model of FISS for proteomic fingerprinting rather than FISS cell cultures include that it is the in vivo 3D model that, in some cases, represents tumour heterogeneity that cannot be observed in vitro. It is also an easy-to-implement model that follows the “3R” (reduction, replacement, refinement) guidelines and resembles spontaneous FISS [ 40 ].…”

“…Fertilised eggs ( Gallus gallus ) ( n = 30) were incubated at 37 °C and 70% humidity in an incubator according to the previously described protocol [ 40 ]. Briefly, on the sixth day of incubation, chick embryos were divided into three groups ( n = 10 per group) and injected with 5 × 10 6 fibrosarcoma cells from the FFS1, FFS3, or FFS5 cell line, and then placed on a CAM silicon ring.…”

“…In veterinary medicine, it has been applied in canine osteosarcoma, canine soft-tissue sarcoma, canine mammary gland tumors, feline mammary carcinoma, and feline fibrosarcoma research [ 35 , 36 , 37 , 38 , 39 ]. In the case of FISS, the CAM model has been shown to closely resembles spontaneous FISS, making it a good alternative for expensive, time-consuming rodent models, which as opposed to the CAM model, require Animal Ethics Commission approval [ 40 ].…”

Resolvin D1 suppresses pannus formation via decreasing connective tissue growth factor caused by upregulation of miRNA-146a-5p in rheumatoid arthritis

“…The chick chorioallantoic membrane (CAM) assay was carried out to evaluate the angiogenic activity in vivo as described previously [22]. Embryonated chicken eggs (~10 per treatment) were incubated at 37°C and treated with RvD1 (0, 20, 100 nM), chicken miRNA-146a-5p agomir or a lentiviral vector harboring RNAi sequence targeting the CTGF gene (Gene Pharma, China) respectively, which were absorbed on sterile Whatman GB/B glass fiber filter disks (6 mm in diameter; Reeve-Angel, Clifton, NJ, USA) on day 7.…”