Functional Analysis of Human Hepatocytes Isolated From Chimeric Mouse Liver

Area of Science:

• Hepatology
• Transplantation Biology
• Virology

Background:

• Developing reliable models for human liver function and disease is crucial.
• Primary human hepatocytes (PHHs) are essential for studying human liver-specific processes.
• Limitations exist in maintaining PHH viability and function in vitro and in vivo.

Purpose of the Study:

• To establish a chimeric mouse model with a humanized liver using PHHs.
• To assess the engraftment, proliferation, and functionality of transplanted human hepatocytes.
• To evaluate the susceptibility of the humanized liver model to hepatitis B virus (HBV) infection.

Main Methods:

• Transplantation of primary human hepatocytes (PHHs) from a Japanese donor into uPA/SCID mice.
• Expansion and purification of human hepatocytes from chimeric mouse liver (hcPHs).
• Second-passaging of hcPHs into recipient mice.
• Analysis of human albumin production and liver-specific enzyme gene expression.
• Infection of chimeric mice with hepatitis B virus.

Main Results:

• Successful engraftment and significant proliferation (over 100-fold) of PHHs in mouse liver.
• Establishment of a secondary humanized liver model through transplantation of hcPHs.
• Demonstrated production of human albumin and preservation of liver-specific enzyme genes by PHHs and hcPHs.
• Successful infection of chimeric mice with HBV, with detectable viral DNA and HBsAg levels.

Conclusions:

• Humanized liver chimeric mice established with PHHs serve as a robust platform for studying human liver biology.
• The model supports sustained human hepatocyte function and is susceptible to HBV infection.
• Purified human hepatocytes (hcPHs) from chimeric mice are a viable cell source for further research and potential therapeutic applications.