Development of orthotopic tumour models using ultrasound-guided intrahepatic injection

Abstract

Mouse models of human diseases are an essential part of the translational pipeline. Orthotopic tumour mouse models are increasingly being used in cancer research due to their increased clinical relevance over subcutaneous xenograft models, particularly in relation to metastatic disease. In this study, we have developed orthotopic colorectal cancer liver metastases (CRCLM) and primary cholangiocarcinoma (CCA) models in BALB/c nude mice using minimally invasive ultrasound-guided intrahepatic injection. Due to its minimally invasive nature, the method reduced risk from surgical complications whilst being fast and easy to perform and resulted in measurable tumour volumes 1 to 3 weeks post-injection. Tumour volumes were monitored in vivo by weekly high-frequency ultrasound (HF-US) and/or twice weekly bioluminescence imaging (BLI) and confirmed with end-point histology. Take rates were high for human CRC cells (>73%) and for CCA cells (90%). We have demonstrated that this method reliably induces CRCLM and CCAs, in which tumour volume can be monitored throughout using HF-US and/or BLI. This provides a promising experimental tool for future testing of cancer therapeutics in an orthotopic model.

CD105 is a prognostic marker and valid endothelial target for microbubble platforms in cholangiocarcinoma

“…Murine subcutaneous xenograft cancer models, though a recognised platform for the assessment of cancer targeting, suffer the detriment of not accounting for the influence of a natural tumour microenvironment, on which orthotopic syngeneic cancer models shed more light [ 31 ]. The engrafting of human CCA xenografts within the liver is however technically challenging, although our group has had recent success in this respect utilizing ultrasound-guided injection into the liver [ 32 ]. In addition, it would have been ideal to conduct our in vivo experiments with a hilar CCA cell line, but the use of extra-hepatic CCA cell lines was favoured due to their availability, ease of xenograft establishment and reproducibility.…”

Evolution of the Experimental Models of Cholangiocarcinoma

“…More recently, a minimally invasive ultrasound-guided intrahepatic injection was established. Due to its nominally invasive nature, this method decreases the risks of surgical complications and provides a potential experimental tool for future screening of cancer therapeutics in orthotopic models [208].…”

The Application Progress of Patient-Derived Tumor Xenograft Models After Cholangiocarcinoma Surgeries

“…Presently, the cholangiocarcinoma (CCA) PDX model establishment method is divided into the subcutaneous transplantation model and orthotopic transplantation model. The methods of establishing subcutaneous transplantation models (28) and the orthotopic transplantation models (29) have been described in detail in the literature. Usually, the F1-F3 generations are used for reproduction, and the F3 generations are used for drug testing.…”

“…This is such a substantial amount of time (86). Moreover, the success rate of orthotopic transplantation is low (40%) (29).…”

“…Improved transplantation methods may be used, such as ultrasound-guided fine needle aspiration. Ultrasound-guided fine needle aspiration technique can also be used for tissue acquisition (29).…”