Urokinase-type plasminogen activator transgenic mouse

Patent metadata

Field	Value
Publication number	US-9955675-B2
Application number	US-201314397091-A
Country	US
Kind code	B2
Filing date	Apr 25, 2013
Priority date	Apr 27, 2012
Publication date	May 1, 2018
Grant date	May 1, 2018

Abstract

The present invention provides a mouse with liver damage, having a high degree of damage against the mouse's original hepatocytes while having a uPA gene in a heterozygous form, and a method for efficiently preparing the mouse. Specifically, the method for preparing a mouse with liver damage having the uPA gene in a heterozygous form comprises the following steps of:
(i) transforming mouse ES cells with a DNA fragment containing a liver-specific promoter/enhancer and cDNA that encodes a urokinase-type plasminogen activator operably linked under the control thereof;
(ii) injecting the transformed mouse ES cells obtained in step (i) into a host embryo;
(iii) transplanting the host embryo obtained in step (ii) via the injection of the ES cells into the uterus of a surrogate mother mouse, so as to obtain a chimeric mouse; and
(iv) crossing the chimeric mice obtained in step (iii), so as to obtain a transgenic mouse in which the DNA fragment is introduced in a heterozygous form.

First claim

The invention claimed is:

1. A chimeric mouse, which has a chimeric liver containing human hepatocytes, wherein human hepatocytes account for at least 10% of all hepatocytes in the chimeric liver, obtained by:
transforming mouse ES cells with a DNA fragment containing a liver-specific promoter/enhancer and cDNA that encodes uPA operably linked under the control thereof;
injecting the transformed mouse ES cells obtained above into a host embryo;
transplanting the host embryo obtained above via the injection of the ES cells into the uterus of a surrogate mother mouse, so as to obtain a chimeric mouse;
crossing the chimeric mice obtained above, so as to obtain a transgenic mouse with liver damage in which the DNA fragment is introduced in a heterozygous form, wherein the serum ALT level increases at least from when it is 6 weeks old to when it is 8 weeks old;
crossing the mouse with liver damage with an immunodeficient mouse forming an immunodeficient mouse with liver damage having the uPA gene in a heterozygous form; and
transplanting human hepatocytes into the immunodeficient mouse with liver damage having the uPA gene in a heterozygous form.

2. A chimeric mouse, which has a chimeric liver containing human hepatocytes, wherein the blood human albumin concentration is 1 mg/mL or higher, obtained by:
transforming mouse ES cells with a DNA fragment containing a liver-specific promoter/enhancer and cDNA that encodes uPA operably linked under the control thereof;
injecting the transformed mouse ES cells obtained above into a host embryo;
transplanting the host embryo obtained above via the injection of the ES cells into the uterus of a surrogate mother mouse, so as to obtain a chimeric mouse;
crossing the chimeric mice obtained above, so as to obtain a transgenic mouse with liver damage in which the DNA fragment is introduced in a heterozygous form, wherein the serum ALT level increases at least from when it is 6 weeks old to when it is 8 weeks old;
crossing the mouse with liver damage with an immunodeficient mouse forming an immunodeficient mouse with liver damage having the uPA gene in a heterozygous form; and
transplanting human hepatocytes into the immunodeficient mouse with liver damage having the uPA gene in a heterozygous form.

Assignees

• Tokyo Metropolitan Inst Medical Science
• Chugai Pharmaceutical Co Ltd
• Phoenixbio Co Ltd

Inventors

• Kohara Michinori
• Jishage Koichi
• Kawase Yosuke
• Mukaidani Chise
• Oshita Hiroki
• Hamamura Satoko

Classifications

• G01N33/5067

  Liver cells · CPC title

• A01K2207/15

  Humanized animals · CPC title

• A01K2217/206

  Animal model comprising tissue-specific expression system, e.g. tissue specific expression of transgene, of Cre recombinase · CPC title

• A01K67/0271

  Chimeric vertebrates, e.g. comprising exogenous cells · CPC title

• A01K2267/035

  Animal model for multifactorial diseases · CPC title