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Bioreactor for constructing tissue engineering liver by using in-vitro circulatory perfusion

A bioreactor and extracorporeal circulation technology, which can be used in specific-purpose bioreactor/fermenter, bioreactor/fermenter combination, tissue cell/virus culture device, etc. Problems such as engineering the liver, unable to completely replace the liver, etc., to achieve the effect of increasing the planting density

Inactive Publication Date: 2015-05-13
昆明市第一人民医院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Since none of the aforementioned bioreactors addresses the issues of oxygen supply and cell growth within a three-dimensional scaffold, their use is limited
At present, these bioreactors can partially replace failed livers in a short period of time, but they cannot completely replace liver functions, nor can they cultivate real tissue-engineered livers (Yu CB, Pan XP, Li LJ. 2009. Progress in bioreactors of bioartificial livers. Hepatobiliary Pancreat Dis Int 8:134-140.)

Method used

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  • Bioreactor for constructing tissue engineering liver by using in-vitro circulatory perfusion

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1: Stem cells cultured on whole liver decellularized scaffolds

[0024] Under sterile conditions, the whole rat liver is decellularized to obtain a biological scaffold with liver network structure and vascular system. The inlet of the stent is the hepatic portal vein, and the outlet is the hepatic vein, and the two ports are respectively connected to the inlet pipe and the outlet pipe of the bioreactor. Connect the other end of the inlet pipe and the outlet pipe of the bioreactor to the culture solution storage tank, and pre-set 106 / ml stem cell suspension in the culture solution storage tank under sterile conditions. The culture solution storage tank, culture solution suspension and liver tissue culture chamber were placed in a carbon dioxide incubator, and the cell suspension was slowly pumped into the whole liver decellularized scaffold by a peristaltic pump, and perfused for 1 cycle. After standing still for one day, the liquid in the culture solution stora...

Embodiment 2

[0025] Embodiment 2: the influence of culture medium on liver growth

[0026] Resect the rat liver under aseptic conditions, rinse the red blood cells on the surface and in the pipeline, measure the liver volume, place it in the liver tissue culture room, connect the hepatic portal vein to the bioreactor inlet pipe, and connect the hepatic vein to the outlet pipe. The other end of the inlet pipe and the outlet pipe of the device are connected with the culture solution storage tank. The culture solution storage tank, culture solution suspension and liver tissue culture chamber were placed in a carbon dioxide incubator, and the liver culture solution was continuously perfused into the rat liver through a peristaltic pump to maintain the pressure at the entrance of the hepatic portal vein at 10 cm water column (approximately Physiological state pressure), measure the size of the liver after two weeks of raising, and observe the influence of the culture medium on the growth of the...

Embodiment 3

[0027] Example 3: Effects of different hydrostatic pressures on liver regeneration

[0028] Resect the rat liver under aseptic conditions, rinse the red blood cells on the surface and in the pipeline, measure the liver volume, place it in the liver tissue culture room, connect the hepatic portal vein to the bioreactor inlet pipe, and connect the hepatic vein to the outlet pipe. The other end of the inlet pipe and the outlet pipe of the device are connected with the culture solution storage tank. The culture solution storage tank, culture solution suspension and liver tissue culture chamber were placed in a carbon dioxide incubator, and the liver culture solution was continuously perfused into the rat liver through a peristaltic pump, and the pressure at the entrance of the hepatic portal vein was maintained at 15 cm of water column for culture (high In the physiological state), the liver size was measured after two weeks of culture, and the effect of high hydrostatic pressure ...

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Abstract

The invention relates to a three-dimensional tissue culture device and a three-dimensional tissue culture method. The bioreactor comprises an in-vitro perfusion circulating system, a negative pressure feedback regulation system, a perfusion flow control module and a liver tissue culture module, wherein the in-vitro perfusion circulating system comprises a closed pipeline system, a tissue perfusion generator and a perfusate suspension update device; the negative pressure feedback regulation system comprises an internal pipeline pressure monitoring system, a computer control module and a perfusion device controlled and regulated by a computer; and the liver tissue culture module comprises a constructed sterile culture room and a constructed three-dimensional tissue culture scaffold. The in-vivo environment can be simulated, a device is provided for culturing tissue engineering organs in vitro, and a tool is provided for research of stem cell differentiation and cell growth under three-dimensional conditions.

Description

technical field [0001] The invention belongs to the field of tissue engineering and biomanufacturing, and relates to (but not limited to) a bioreactor for constructing a tissue engineered liver by three-dimensional culture of liver tissue. It can realize cell planting, tissue culture perfusion and extracorporeal circulation in the process of tissue engineering liver in vitro culture, and simulate blood circulation under physiological conditions, and the main parameters can be adjusted. Background technique [0002] The liver is an important detoxification and digestive organ of the human body. End-stage liver diseases such as hepatitis, cirrhosis, and severe liver failure have seriously affected people's quality of life and become one of the diseases with the highest mortality in the world. Currently, liver transplantation is recognized as the only effective treatment. In China alone, about 300,000 patients with end-stage liver disease are waiting for liver transplantation,...

Claims

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Application Information

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IPC IPC(8): C12M3/00C12M1/36C12M1/34C12M1/12C12M1/04
Inventor 张蕾张涛
Owner 昆明市第一人民医院
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