Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

New application of immunodeficient mice model

A mouse model, defect-type technology, applied in the field of biomedicine, can solve problems such as prone to rejection, achieve controllable induction of liver damage, increase expression, and high degree of immunodeficiency

Pending Publication Date: 2018-09-07
湖南昭泰生物医药有限公司
View PDF7 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] CN 102460162A discloses a method for expanding human hepatocytes in vivo, the method comprising transplanting human hepatocytes into immunodeficient (Rag - / - deficient, B6 mouse background) and Fah-deficient mice that were also deficient in IL-1R or were given an IL-1R antagonist and allowed the hepatocytes to expand, however, the mouse model of the invention Need IL-1R deficiency or need IL-1R antagonist, strong immunity, prone to rejection after transplantation of human hepatocytes

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • New application of immunodeficient mice model
  • New application of immunodeficient mice model
  • New application of immunodeficient mice model

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0071] Example 1 In vivo expansion of human primary hepatocytes

[0072] (1) In vitro isolation of human primary cells

[0073] The human primary liver parenchyma tissue of fresh donor was separated by collagenase two-step perfusion method. The specific method was as follows: firstly, the fresh liver tissue was added with 0.4mM ethylene glycol bis(2-aminoethyl ether)tetraacetic acid under sterile conditions. (EGTA) in PBS buffer perfusion for 15 minutes (flow rate of 5mL / min); followed by adding 0.1mg / mL collagenase IV and 2mM Ca 2+ Continue to perfuse the PBS buffer solution for 15 minutes (flow rate is 2mL / min); after the digestion is complete, use ophthalmic tweezers to gently shake the liver cells to release a single liver cell, and continue to digest the undigested tissue with 0.1mg / mL collagenase IV for 20 minutes. Minutes, a large number of hepatocytes were obtained; the above cells were centrifuged with DMEM medium supplemented with 10% fetal calf serum at a speed of ...

Embodiment 2

[0082] Example 2 Mouse liver detection after transplantation of human hepatocytes

[0083] (1) Flow detection

[0084] Take mouse liver tissue and add antibody blocking receptors, add fluorescently labeled antibodies HLA-A / B / C and MHC-I under dark conditions, wash with PBS buffer twice after labeling, and then perform flow detection.

[0085] The result is as Image 6 As shown, after stopping the addition of NTBC, the proportion of human hepatocytes in mouse liver cells increased significantly.

[0086] (2) H&E staining

[0087] The mouse liver organs were fixed in neutral formic acid solution overnight, dehydrated with increasing concentrations of ethanol solutions, became transparent in xylene, dipped in wax, embedded to form wax blocks of appropriate size; cut the tissue with a microtome After slicing into 4-6μm sections, carry out dewaxing and hydration treatment; the hydrated sections are stained with hematoxylin, rinsed with running water, stained with eosin, rinsed wit...

Embodiment 3

[0092] Example 3 In vivo tracing of human primary hepatocytes

[0093] (1) Cell culture dish pre-coated

[0094] Use 0.1% gelatin (Gelatin) to pre-coat the culture dish or culture plate, specifically: dissolve 1mL 1% gelatin in 10mL DMEM medium, add it to a 60mm culture dish, shake gently to make the liquid cover the entire bottom of the dish, place Place in a 37°C incubator for 4 hours, take out the culture dish, suck off the excess supernatant, add DMEM medium to wash twice, and set aside.

[0095] (2) Culture of primary human hepatocytes

[0096] Isolate donor hepatocytes and adjust the cell concentration to 5×10 5 / cm 2 , inoculated on Gelatin-coated 60mm culture dishes, the culture medium was DMEM medium supplemented with 10% FBS+1% double antibody+10nM / mL hepatocyte growth factor (HGF); placed in 5% CO 2 After culturing in a 37°C incubator for 4 hours, replace with fresh medium to remove some unattached cells; replace with fresh medium every 2 days.

[0097] Human ...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention provides new application of an immunodeficient mice model. NOD (Non-Obese Diabetic) / SCID (Severe Combined Immune Deficiency) / IL2rg<- / -> (Interleukin 2 Receptor, Gamma <- / ->) / Fah<- / -> (Fumarylacetoacetate Hydrolase <- / ->) mice can be used for carrying out in-vivo amplification on human cells. The NOD / SCID / IL2rg<- / -> / Fah<- / -> mice are high in immunodeficient degree and controllable inliver injury induction, rejection reaction cannot be generated after human hepatocytes are transplanted, and the NOD / SCID / IL2rg<-> / Fah<- / -> mice can be used as an in-vivo amplification and in-vivo tracking model of the human hepatocytes.

Description

technical field [0001] The invention belongs to the field of biomedicine and relates to a new application of an immunodeficiency mouse model, in particular to a NOD / SCID / IL2rg - / - / Fah - / - Application of a mouse model for in vivo expansion and in vivo tracking of human hepatocytes. Background technique [0002] The liver is the largest and most important organ for biochemical reactions in the human body, and it is also the main target organ for drug damage. Nearly 1000 drugs have been found to be associated with liver injury, the most common single reason for drug withdrawal from the market. More than half of acute liver failure in the United States is caused by drugs, the most common being Paracetamol, acetaminophen or acetaminophen (AAP). The annual incidence of AAP-associated acute liver failure has increased from 28% in 1998 to 51% in 2003. In cases of acute liver failure not caused by AAP, antibacterial drugs, especially anti-tuberculosis drugs, are the main drugs f...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): C12N5/071C12N15/867A61K49/00
CPCA61K49/0017C12N5/067C12N5/0679C12N5/0686C12N5/0688C12N15/86C12N2509/00C12N2740/15043
Inventor 不公告发明人
Owner 湖南昭泰生物医药有限公司
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com