Method for establishing three-dimensional tissue in vitro

A three-dimensional, tissue technology, applied in the direction of prosthesis, medical science, etc., can solve the problem of inconvenient way of obtaining materials

Inactive Publication Date: 2015-01-14
SHANGHAI SIXTH PEOPLES HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] For the seed cells used in urethral tissue engineering, the most common traditional methods are urothelial cells a...

Method used

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  • Method for establishing three-dimensional tissue in vitro

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 14

[0026] Example 14-layer small intestinal mucosa acellular matrix (SIS) scaffold combined with tongue epithelial cells and skeletal muscle cells in vitro to construct three-dimensional urethral tissue

[0027] 1. Stent disinfection

[0028] The 4-layer SIS scaffold purchased from COOK Company of the United States was cut into small pieces with a size of 2 cm × 3 cm, sterilized with ethylene oxide, and packaged and stored at room temperature for later use.

[0029] 2. Drawing materials

[0030] Patients with anterior urethral stricture who underwent tongue mucosa replacement urethroplasty were selected. When they underwent tongue mucosa replacement urethroplasty, the remaining tongue mucosa tissue was collected.

[0031] 3. Seed cell culture

[0032] 1) Culture of tongue mucosal epithelial cells: Remove the tongue mucosa tissue, remove the serosal layer tissue, put it into a 50ml centrifuge tube containing chloramphenicol and shake for 6-9 times, wash with PBS for 3 times, and...

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Abstract

The invention provides a method for establishing a three-dimensional tissue in vitro. The method comprises the steps of separately culturing epithelial cells and muscle cells; implanting a small intestinal submucosa acellular matrix stent into a muscle cell suspension; continuing to culture after finishing oscillating and compounding operations; and dropwise adding an epithelial cell suspension on a muscle cell layer of the stent treated in the step 2, and continuing to culture to obtain the three-dimensional tissue. According to the method, the epithelial cells and the muscle cells of tongue mucosa are used as seed cells, so that materials are conveniently obtained, the cells can be increased to the relatively high quantity within relatively short time, and therefore, the application requirement is met; the muscle cells are compounded with the small intestinal submucosa acellular matrix stent by using an oscillating method, so that a plurality of muscle cells can be soaked into the stent and can be relatively uniformly distributed; meanwhile, the stent can be loosened as much as possible on the premise that the stent is not obviously damaged, so that nutrient permeation and neovascularization can be favorably realized; and the three-dimensional tissue which can be used for tissue engineering urethra can be established.

Description

technical field [0001] The invention relates to the technical field of tissue engineering, in particular to a method for constructing a three-dimensional tissue in vitro, and the three-dimensional tissue constructed by the method. Background technique [0002] Surgical repair of congenital or acquired urethral diseases is one of the difficult problems that are constantly being explored in the field of urology. The development of tissue engineering technology provides a new idea for the repair of urethral stricture. Among them, acellular matrix is ​​an important content of tissue engineering as a scaffold for cell regeneration. At present, the most studied acellular matrix is ​​small intestinal submucosal acellular matrix (SIS), which has been applied to clinical urethral repair and has achieved good results. [0003] The submucosal acellular matrix of the small intestine is a kind of allogeneic tissue obtained from the small intestine of pigs. The mucosal layer, serosa lay...

Claims

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Application Information

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IPC IPC(8): A61L27/38A61L27/36
Inventor 徐月敏吕向国冯超
Owner SHANGHAI SIXTH PEOPLES HOSPITAL
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