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Matrix, cell implantation and method for their production and use

A graft, matrix technology, applied in the fields of drug combination, medical science, digestive system, etc., can solve the problem of not getting organ replacements

Inactive Publication Date: 2006-08-09
HUMANAUTOCELL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] However, the biological matrices produced so far using this method have not been satisfactory in all cases, especially with regard to the ability of grafts built on these matrices to be accepted and function
In particular, to date no acceptable organ replacements have been obtained using liver and pancreas transplants

Method used

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  • Matrix, cell implantation and method for their production and use

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0077] Prepare matrix

[0078] a) Polymer-free solution

[0079]Polymer pellets (Resomer® RG 858 from Boehringer, Ingelheim) were frozen in liquid nitrogen and crushed in the frozen state (Döschle extrusion system; 12000 rpm, 2 min). The broken polymer particles are screened. Particles ranging in size from 108 μm to 250 μm were used to prepare the matrices. In this regard, 16% by weight of polymer employed has a particle size between 108 μm and 140 μm, while 22% by weight of polymer used has a particle size between 145 μm and 180 μm and 46% by weight of polymer , the particle size is between 185 μm and 220 μm, and 16% by weight of polymer is used, and the particle size is between 225 μm and 250 μm. Sodium chloride (common salt) was screened, and sodium chloride particles with a particle size of 250 μm to 425 μm were used to prepare the matrix. In this regard, 25% by weight of the salt used has a particle size between 250 μm and 320 μm, 44% by weight of the salt used has a ...

Embodiment 2

[0087] a) Coating the matrix with fibronectin

[0088] The matrix from Example 1 was immersed in a carbonate buffer solution containing 3 μg human plasma-derived fibronectin (Sigma) / ml and having a pH of 9.4. After approximately 60 s, the matrix was removed from solution, lyophilized and gamma-irradiated.

Embodiment 3

[0090] cell separation

[0091] A part of the liver is removed from the individual to be transplanted in a manner known per se. The removed liver part was first perfused with solution (8.3g NaCl; 0.5g KCl; 2.38g HEPES; made up to 1000ml with distilled water, pH7.4) at 37°C for 7 minutes at a flow rate of 30ml / min. The liver fraction was then treated with a collagenase / trypsin inhibitor solution (8.3 g NaCl; 0.5 g KCl; 2.38 g HEPES; 0.7 g CaCl) at 37°C at a flow rate of 30 ml / min. 2 ×2H 2 O; 500 mg collagenase (Collagenase H, Boehringer Mannheim, Mannheim, Germany); 7.5 mg trypsin inhibitor (ICN, Eschwege, Germany; make up to 1000 ml with distilled water, pH 7.35) perfuse for an additional 6 to 7 minutes . After infusion, liver sections were dissected and cultured in Williams medium E with careful shaking. The cell suspension was filtered (nylon mesh, 200 μm) and washed with Williams medium E. Afterwards, the cells were centrifuged at 50g for 3 minutes at 4°C. Cell viabil...

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Abstract

The invention relates to porous matrices based on a biologically compatible polymer or polymer mixture, to a cell implantation that is established on said matrices and to additional cell implantation based on cell mixtures of hepatocytes and islets of Langerhans. The invention also relates to a method for producing porous matrices, to matrices obtained according to said method and to a special method for obtaining cells for the inoculation of an implantable matrix.

Description

field of invention [0001] The present invention relates to porous matrices based on bioresistant polymers or polymer mixtures, to cell grafts built thereon, to other cell grafts based on cell mixtures consisting of hepatocytes and pancreatic islets, to methods for preparing porous matrices The method and use of the method to obtain a matrix, and relates to specific methods for obtaining cells for seeding a transplantable matrix. technical background [0002] Tissue engineering is an interdisciplinary field that combines engineering and materials science with medicine. The goal is to restore damaged tissue or improve its function. [0003] The principle of tissue engineering is extremely simple: First, some cells are taken from the patient and propagated in vitro. The multiplied cells are then implanted into the scaffolding material, resulting in a complete living tissue substitute, which is then reimplanted into the patient. In contrast to traditional allogeneic transplan...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C08J9/26A61L27/56A61L27/58A61L27/50A61L27/18A61L27/26A61L27/38C12N5/07C12N5/071
CPCA61L27/3895A61L27/56A61L27/58A61L27/3886A61L27/3804A61L27/26A61P1/16A61P1/18A61P3/10A61P35/00
Inventor M·格尔纳P·M·考夫曼
Owner HUMANAUTOCELL