Establishment of Liver Cancer Patient-derived Xenograft Models

Summary

Here, we present a protocol to establish liver cancer patient-derived xenograft models for the preclinical study of novel anticancer drugs.

Abstract

Patient-derived xenograft (PDX) models are established by transplanting immune-compromised mice with tumor samples from cancer patients. The application of PDX models has facilitated the development of anticancer drugs in preclinical studies. In this article, we present a method to establish a liver cancer PDX model. Human liver cancer tissues are subcutaneously injected into scid mice to generate a bank of tumors, which are subsequently passaged into different generations of mice to maintain the liver cancer PDX models. The liver cancer PDX models mostly resemble their original tumor properties as determined by immunohistochemistry analysis and western blot assay. Treatment with sorafenib, a Food and Drug Administration (FDA)-approved standard first-line drug that has been used for the treatment of unresectable liver cancers, suppresses the tumor growth in the liver cancer PDX model. Although there are limitations to this liver cancer PDX model, it has helped scientists to investigate, in preclinical studies, novel therapies for liver cancer treatment which are more precise and clinically relevant.

Introduction

PDX models are established by transplanting immune-compromised mice with tumor samples from cancer patients^1,2. PDX models have been established in various types of cancers, including breast cancer, lung cancer, liver cancer, pancreatic cancer, and so on³. PDX models retain the genomic, histologic, and biological properties of the corresponding primary tumors. More importantly, the response of PDX models after anticancer drug treatment has been found to be associated with the clinical outcome of cancer patients, which is important for healthcare professionals when making therapeutic dec....

Protocol

This protocol has been conducted at the University of Hong Kong with approval from the Institutional Review Board of the University of Hong Kong/Hospital Authority of Hong Kong (UW05-3597/I022).

1. Preparation of the Patients’ Tumor Sample (~2 cm x 2 cm)

1. Prepare Hank’s balanced salt solution (HBSS; 140 mM NaCl, 5 mM KCl, 1 mM CaCl₂, 0.4 mM MgSO₄·7H₂O, 0.5 mM MgCl₂·6H₂O, 0.3 mM Na₂HPO₄.......

Discussion

Liver cancer has very a low survival rates and a high probability of metastasis, making it one of the most aggressive cancers. In this article, we described a detailed protocol for the generation of an improved liver cancer PDX model. In this model, patient-derived tumors were injected into scid mice, passaged, and subsequently applied in the evaluation of the anticancer effects of sorafenib. More importantly, this liver cancer F1 PDX model mostly retains the characteristics of the original clinical tumor, has similar dr.......

Materials

Name	Company	Catalog Number	Comments
1.5 ml microcentrifuge tube	Pipette	22363204
Ammonium chloride	Sigma-Aldrich	254134
Anesthesia bix	Patterson Veterinary
Anesthesia machine	Patterson Veterinary
Buprenorphine	Sigma-Aldrich	B-044
Calipers	Flowler	54-100-167
Collagenase IV	Thermo Fisher Scientific	17104019
DMEM/F12	Thermo Fisher Scientific	11320033
Fetal bovine serum	Sigma-Aldrich	F2442
Forceps	Roboz	RS-5135
Gelatin solution	Sigma-Aldrich	G1393
Hank's Balanced Salt Solution	Thermo Fisher Scientific	14025076
Isoflurane	Vet one	1038005
Penicilin	Sigma-Aldrich	13752
Scissors	Roboz	RS-5881
Streptomycin	Sigma-Aldrich	S6501
Trocars	Innovative Research of America	MP-182
Weight scale	Ohuas	Scout Pro SP601