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Tissue-engineered neural tissues and construction method thereof

A technology of nerve tissue and tissue engineering, applied in the direction of nervous system cells, medical science, prosthesis, etc., to achieve the effect of increasing pore size, promoting development, and promoting differentiation

Inactive Publication Date: 2011-11-02
INST OF BASIC MEDICAL SCI ACAD OF MILITARY MEDICAL SCI OF PLA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The application of liquid type I collagen / Matrigel in neural tissue engineering has not been reported as a scaffold material for neural stem cells

Method used

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  • Tissue-engineered neural tissues and construction method thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0021] Example 1 In Vitro Construction of Tissue Engineered Nerve Tissue

[0022] Reagents required for cell isolation, culture and tissue identification used in this example:

[0023] (1) Dissection solution: 3g D-glucose, 7.5g sucrose, 2.34g HEPES, 8g NaCl, 0.4g KCl, 0.24g Na 2 HPO 4 , 4.12 g H 2 O, 0.03g KH 2 PO 4 Add it to 1L of ultrapure water, adjust the pH value to 7.2-7.4, filter and sterilize with a 0.22 μm filter membrane, and store at 4°C.

[0024] (2) DMEM-F12 medium: Dissolve one package of DMEM-F12 dry powder medium in 1000mL ultrapure water, add 1.2g NaHCO 3 , adjusted the pH value to 7.2-7.4, sterilized by filtration through a 0.22 μm filter membrane, aliquoted and stored at 4°C.

[0025] (3) Concentrated medium (10×H-DMEM): Dissolve one package of DMEM-F12 dry powder medium in 100mL ultrapure water, add 1.2g NaHCO 3 , adjusted the pH value to 7.2-7.4, sterilized by filtration through a 0.22 μm filter membrane, aliquoted and stored at 4°C.

[0026] (4) ...

Embodiment 2

[0034] Example 2 Live / Dead and Immunofluorescence Detection of Tissue Engineered Nerve Tissue Constructed in Vitro

[0035] Live / Dead detection: for the tissue engineered neural tissue constructed in vitro in Example 1, on the 1d, 3d, and 7d of culture, the Live / Dead kit was used to detect the effect of cell survival in the tissue engineered neural tissue. The method is as follows: first, add 20 μl of 2mM EthD-1 and 5 μl of 4mM Calcein AM to 10ml of PBS to prepare the dye solution; discard the medium, rinse with PBS for 3 times; add 1mL of the dye solution, and stain at 37°C for 60min; discard the dye solution, fresh Wash again with PBS for 30 minutes; randomly select 3-5 fields of view and use laser confocal to detect cell survival.

[0036] The results showed that the neural stem cells and differentiated cells survived well in the collagen / Matrigel complex, and the survival rate was still as high as 87% when cultured in vitro for 7 days.

[0037] Immunofluorescence detectio...

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Abstract

The invention discloses tissue-engineered neural tissues and a construction method thereof, belonging to the technical field of tissue engineering and regeneration medicine. Subcultured and purified neural stem cells serve as seed cells of the tissue-engineered neural tissues, a liquid type I collagen and Matrigel matrix serves as stent materials, and the seed cells and the stent materials are cultured after being compounded in vitro. A stent system adopted in the invention can be used for effectively maintaining the survival and proliferation of the neural stem cells, promoting the differentiation of the neural stem cells, and forming engineered neural tissues consisting of mature neurons, astrocytes and oligodendrocytes. The tissue-engineered neural tissues have an important significance in treating neural system injury through the transplantation of the tissue-engineered neural tissues, and can also serve as an in-vitro model for the psychological research of neurodevelopment, cerebral trauma and neuroelectricity.

Description

technical field [0001] The invention belongs to the technical field of tissue engineering and regenerative medicine, and in particular relates to a tissue-engineered nerve tissue and a construction method thereof. Background technique [0002] Tissue engineering is a new interdisciplinary subject developed in the late 1980s. In recent years, with the development of life science and material science, researchers have made great achievements in the research of bone, cartilage, skin and other tissues and organs in vitro. Breakthrough progress, among them, some tissue engineering products such as cartilage and skin have been commercialized. Although due to the complexity of the nervous system itself and the late start of research on tissue-engineered neural tissue, the neural tissue constructed in vitro is far from being commercialized; however, with the research on stem cells, neurotrophic factors and biomaterials With the rapid development of the field, in vivo transplantatio...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61L27/38C12N5/0797
Inventor 王常勇李蓝兰林秋霞陈威震韩尧张志立王妍
Owner INST OF BASIC MEDICAL SCI ACAD OF MILITARY MEDICAL SCI OF PLA
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