Generation of Subcutaneous and Intrahepatic Human Hepatocellular Carcinoma Xenografts in Immunodeficient Mice

Sharif U. Ahmed; Murtuza Zair; Kui Chen; Matthew Iu; Feng He; Oyedele Adeyi; Sean P. Cleary; Anand Ghanekar

doi:10.3791/50544

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Method Article

Generation of Subcutaneous and Intrahepatic Human Hepatocellular Carcinoma Xenografts in Immunodeficient Mice

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DOI:

10.3791/50544

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September 25th, 2013

Sharif U. Ahmed¹ , Murtuza Zair¹ , Kui Chen¹ , Matthew Iu¹ , Feng He¹ , Oyedele Adeyi² , Sean P. Cleary³ , Anand Ghanekar¹^,³

¹Toronto General Research Institute, University Health Network, ²Department of Pathology, University Health Network, ³Division of General Surgery, University Health Network

Summary

Human tumor xenografts in immunodeficient mice are valuable tools to study cancer biology. Specific protocols to generate subcutaneous and intrahepatic xenografts from human hepatocellular carcinoma cells or tumor fragments are described. Liver regeneration induced by partial hepatectomy in recipient mice is presented as a strategy to facilitate intrahepatic engraftment.

Abstract

In vivo experimental models of hepatocellular carcinoma (HCC) that recapitulate the human disease provide a valuable platform for research into disease pathophysiology and for the preclinical evaluation of novel therapies. We present a variety of methods to generate subcutaneous or orthotopic human HCC xenografts in immunodeficient mice that could be utilized in a variety of research applications. With a focus on the use of primary tumor tissue from patients undergoing surgical resection as a starting point, we describe the preparation of cell suspensions or tumor fragments for xenografting. We describe specific techniques to xenograft these tissues i) subcutaneously; or ii) intrahepatically, either by direct implantation of tumor cells or fragments into the liver, or indirectly by injection of cells into the mouse spleen. We also describe the use of partial resection of the native mouse liver at the time of xenografting as a strategy to induce a state of active liver regeneration in the recipient mouse that may facilitate the intrahepatic engraftment of primary human tumor cells. The expected results of these techniques are illustrated. The protocols described have been validated using primary human HCC samples and xenografts, which typically perform less robustly than the well-established human HCC cell lines that are widely used and frequently cited in the literature. In comparison with cell lines, we discuss factors which may contribute to the relatively low chance of primary HCC engraftment in xenotransplantation models and comment on technical issues that may influence the kinetics of xenograft growth. We also suggest methods that should be applied to ensure that xenografts obtained accurately resemble parent HCC tissues.

Introduction

Hepatocellular carcinoma (HCC) is the fifth most common cancer worldwide and the most rapidly increasing cause of cancer death in North America. The most prevalent risk factor for HCC is liver cirrhosis, most frequently occurring due to chronic viral hepatitis, alcohol misuse, autoimmune disease, or hereditary metabolic disorders ¹.

Despite the heavy disease burden imposed by HCC on populations worldwide, the pathophysiology of HCC is relatively poorly understood in comparison to other common cancers such as colorectal, breast, or prostate cancer. For example, specific molecular and cellular events driving tumorigenesis remain to....