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Method for constructing and implanting organization engineering liver implant

A construction method and tissue engineering technology, applied in medical science, prosthesis, etc., can solve problems such as difficulty in providing functional support, rapid inactivation, and inability to induce liver parenchyma, so as to promote effective contact, improve implantation, and facilitate diffusion and the effect of nutrient transport

Inactive Publication Date: 2009-09-16
GENERAL HOSPITAL OF PLA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although this method combines the inducing factor, because the inducing factor is only simply compounded with the bioprotein glue, it will be rapidly inactivated after implantation in the body, and cannot induce a sufficient amount of liver parenchyma, making it difficult to provide effective functional support.

Method used

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  • Method for constructing and implanting organization engineering liver implant
  • Method for constructing and implanting organization engineering liver implant

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Example 1 Construction of engineered liver graft unit

[0023] Dissolve the growth factors in 5% agarose, the final concentrations of the growth factors HGF and VEGF are 5 μg / ml respectively, after the agarose coagulates, mechanically break it into small microspheres to obtain agarose Slow-release microspheres.

[0024] Then operate on ice, compound the gel sustained-release microspheres and hydrogel materials to construct a bioactive scaffold; mix liquid type I collagen (from SD rat tail tendon) and 2×DMEM (Sigma, US) in equal proportions, Adjust the pH to 7.4 with 0.1mol / L NaOH solution to obtain a mixture, add gel slow-release microspheres to construct a bioactive scaffold.

[0025] Primary isolated liver cells (SD rats, 200-250 g) were mixed with bioactive scaffolds (on ice) to create liver cell / scaffold complexes. Aspirate the liver cell / scaffold complex into a 1 ml disposable syringe and place at 37 °C, 5% CO 2 Incubate in an incubator for 15 minutes, push out ...

Embodiment 2

[0027] The prepared liver unit was subcutaneously implanted to construct engineered liver tissue. The experiment was carried out as follows:

[0028] Liquid type I collagen (from the tail tendon of SD rats) and 2×DMEM medium were mixed in equal proportions, and the pH value of the mixture was adjusted to 7.2-7.4 with 0.1 mol / L NaOH. Freshly isolated hepatocytes were combined with the above mixture to construct hepatocyte / collagen gel complexes, and placed at 37°C, 5% CO 2 Incubate for 10 minutes in the incubator. After the complex forms a gel, it is broken up into small pieces of hepatic cell units. After creating a subcutaneous pouch in the armpit of the rat, hepatocyte units were implanted into the pouch.

[0029] The results of in vitro experiments show that after the hepatocytes are compounded with liquid type I collagen, the cells are evenly distributed in the whole complex and can grow in three dimensions; during the whole in vitro culture process, the hepatocytes alw...

Embodiment 3

[0031] Construction of engineered liver tissue in situ in the liver using bone marrow stromal stem cells as seed cells

[0032]Bone marrow stromal stem cells of SD rats cultured to the fourth generation by conventional methods were compounded with liquid type I collagen, concentrated DMEM (2×) culture medium and commercially available hepatocyte culture medium, and growth factors HGF and VEGF were added, and the final concentrations were respectively 5 μg / ml. Aspirate the complex into a 1ml syringe and place it in a 37°C, 5% CO2 incubator for 15 minutes, push it out after the complex forms a semi-solid gel, and cut it into small pieces with a length of 1mm to form A cylindrical liver graft unit with a diameter of 0.5 mm and a height of 1 mm.

[0033] The constructed liver graft unit was implanted into the liver of syngeneic rats by injection. One week later, liver-like tissue was formed in the transplanted area. Histological and immunohistochemical tests can be seen; 1) isl...

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Abstract

The invention relates to field of regeneration medical science and organization engineering, and provides an implantable engineering liver implant and construction method thereof. The method includes steps of employing hydrogel material as biological activity support, compounding slow release microsphere containing angiogenesis cell factor and liver cell induction factor (and biological activity substance), compounding liver cell, constructing engineering liver cell / support compound, promoting propagation of liver cell and blood vascularization of engineering liver implant, and after the liver cell / support compound is changed to gel, thrashing the gel to liver implant unit. By implanting the liver implant unit to liver outer site or implanting multi-site to liver, the invention overcomes ischemic problem of engineering liver organization in initial stage of implantation, effectively solves problem of regeneration of engineering liver organization in vivo, and provides a new method for treating various hepatopathies by employing organization engineering method to construct engineering liver organization in vivo.

Description

technical field [0001] The invention relates to the fields of regenerative medicine and tissue engineering, in particular to the construction of an implantable tissue engineered liver graft and its implantation method. Background technique [0002] The liver is the largest internal organ in the human body. It is responsible for important and complex physiological functions and plays an important role in maintaining life and stabilizing the internal environment. There are more than 300 million liver disease patients in the world, one-third of them are in my country, which is one of the regions with the highest incidence of end-stage liver disease in the world. Liver transplantation is currently an effective treatment for acute and chronic liver failure and various hereditary metabolic disorders. However, the widespread application of this technique is limited by the severe shortage of donor organs. The existing extracorporeal artificial liver assisting device can be used as...

Claims

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Application Information

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IPC IPC(8): A61L27/38A61L27/52A61L27/54A61F2/00A61L27/26A61L27/24A61L27/22A61L27/20
Inventor 徐迎新赵云山张博峰李力刘巨超李荣
Owner GENERAL HOSPITAL OF PLA
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