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Method for preparing fusiform complicated organ precursor by using rotary assembling die

A combined mold and spindle-shaped technology is used in the field of preparing tissue and organ precursors by using synthetic polymer materials and cell matrix materials. Effect

Inactive Publication Date: 2013-01-16
TSINGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The purpose of the present invention is to use the method of rotating combined moulds, to prepare spindle-shaped complex organ precursors, so as to overcome the shortcomings in the field of tissue engineering that cells are difficult to infiltrate into three-dimensional scaffolds, and scaffolds containing various cells are not easy to form; at the same time, molds are avoided. The influence of internal shape and surface roughness on forming makes the forming process more stable and reliable

Method used

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  • Method for preparing fusiform complicated organ precursor by using rotary assembling die
  • Method for preparing fusiform complicated organ precursor by using rotary assembling die
  • Method for preparing fusiform complicated organ precursor by using rotary assembling die

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] 1) Use brass to prepare the bottom mold with three steps, the inner ring mold and the outer ring mold, and nylon to prepare the inner mold with first-level branches; 2) Prepare 1% fibrinogen solution, and insert the bottom of the inner mold into the base mold In the array orifice plate, the array orifice plate is embedded in the bottom of the base mould, the first-level inner ring mold is placed on the first step of the base mould, and fibrinogen and endothelium are injected between the inner mold and the first-level inner ring mould. Mixture of cells at a cell density of 1 x 10 7 cells / mL, make the base mold, inner mold and array well plate rotate together (the speed is 5r / min) and keep the first-stage inner ring mold still, so that the cell matrix solution forms a semi-spindle shape, and use coagulation during the rotation Soak the formed product in enzyme solution (20IU / mL) for 2 minutes to polymerize, remove the first-level inner ring mold, and form a stable first l...

Embodiment 2

[0052] Example 2: (1) Prepare the bottom mold with four steps, the inner ring mold and the outer ring mold with silicon rubber, and prepare the inner mold with secondary branches with polyethylene; (2) prepare 5% fibrinogen solution, Insert the bottom of the inner mold into the array orifice plate of the base mould, the array orifice plate is embedded in the bottom of the base mold, put the first-level inner ring mold on the first step of the base mold, and the inner mold and the first-level inner ring mold Fibrinogen / endothelial cell mixture containing 1% paclitaxel (cell density 1×10 7 pcs / mL), the base mold, the inner mold and the array plate are rotated together (rotating speed is 10r / min) and the first-stage inner ring mold is kept still, so that the cell matrix solution forms a semi-spindle shape, during the rotation process, use Soak the formed product in thrombin solution (10IU / mL) for 1 minute to polymerize, remove the first-level inner ring mold, and form a stable fi...

Embodiment 3

[0053] Example 3: (1) Use polytetrafluoroethylene to prepare a bottom mold with five steps, an inner ring mold and an outer ring mold, and polyurethane to prepare an inner mold with three-level branches; (2) prepare a 10% collagen solution, and The bottom of the inner mold is inserted into the array hole plate of the base mold, the array hole plate is embedded in the bottom layer of the base mold, the first-level inner ring mold is set on the first step of the base mold, and then 1% sodium citrate collagen / endothelial Cell mixture (cell density is 1 x 10 7per mL) into the gap between the inner mold and the first-stage inner ring mold, so that the base mold, the inner mold and the array orifice plate rotate together (the speed is 20r / min) and the first-stage inner ring mold remains constant Rotate to make the cell matrix solution form a semi-spindle shape. During the rotation process, use physical cross-linking method to place it at 37°C for 10 minutes to stabilize the structur...

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Abstract

The invention provides a method for preparing a fusiform complicated organ precursor by using a rotary assembling die, which comprises the following steps of: firstly, preparing a cellular matrix solution and a synthetic macromolecular solution, assembling the assembling die with a multilevel branching internal die, pouring the cellular matrix solution between an internal ring die and a base die, rotating the base die and carrying out physical or chemical cross-linking or polymerization to obtain a cellular matrix layer; secondly, pouring the synthetic macromolecular solution between the cellular matrix layer and an external ring die, rotating and extracting the base die with a cellular culture solution or PBS (Phosphate Buffer Solution) to form an outer support, removing the assembling die to obtain a semi-fusiform three-dimensional structure with multiple branching channels containing different cells and synthetic macromolecule supports, and permeably adhering and mixing the two semi-spindles through either natural or synthetic macromolecular solution to form the complete fusiform complicated organ precursor with the internal branching pipe. The method for preparing the fusiform complicated organ precursor by using the rotary assembling die overcomes the defects that the existing cells are difficult to penetrate into the three-dimensional supports and the supports containing multiple cells are unlikely to form and the like.

Description

technical field [0001] The invention belongs to the technical field of artificial manufacturing of complex tissues and organs of organisms, and particularly relates to a process method for preparing tissue and organ precursors by using synthetic polymer materials and cell matrix materials, which is beyond the current technical field of tissue engineering. Background technique [0002] Tissue engineering was officially proposed and determined by the National Science Foundation of the United States in 1987. It is the application of the principles of cell biology, biomaterials and engineering to research and develop the structure and function of human diseased tissues or organs. The science of biologically active substitutes. Wolter formally proposed the term "tissue engineering" in 1984, and in 1987, the National Science Foundation of the United States formally confirmed that tissue engineering became a new discipline. [0003] In today's medical process, organ transplantatio...

Claims

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Application Information

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IPC IPC(8): A61F2/02A61L27/38A61L27/24A61L27/20A61L27/18
Inventor 王小红黄源文
Owner TSINGHUA UNIV
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