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Method for preparing tissue engineering blood vessel based on 3D bioprinting technology

A tissue engineering and bioprinting technology, applied in the field of bioengineering, can solve the problems of low cell density, low cell compatibility and tissue compatibility of replacement blood vessels, insufficient mechanical strength, etc., achieve good biocompatibility, improve cell Compatibility and histocompatibility, the effect of overcoming the lack of mechanical strength

Active Publication Date: 2015-04-08
江苏知聚知识产权服务有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method has the following disadvantages: the hydrogel has insufficient mechanical strength as a scaffold material, low cell density, low cytocompatibility and histocompatibility for replacing blood vessels

Method used

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  • Method for preparing tissue engineering blood vessel based on 3D bioprinting technology

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Cell handling:

[0047] 1) Take out the culture dish containing human neonatal dermal fibroblasts (hNDFs) from the incubator, wash with phosphate buffer saline (PBS), and digest the cells with trypsin;

[0048] 2) The treated cells were resuspended in 4 mL of cell culture medium and placed in a 10 mL tissue culture flask;

[0049] 3) The flask is placed on a rotary shaker in a volume concentration of 5% CO 2 Environment, cultivated at 37°C for 1 hour, and centrifuged at 3500rpm;

[0050] 4) Use a 3D printer to extrude the cell column on a special, non-adhesive polytetrafluoroethylene or agarose base, and culture it overnight to mature the cell column.

[0051] Pluronic F127 (PF127) processing:

[0052]PF127 with 20%-30% W / W is used, wherein, W / W is: solute mass percent concentration (w / w) = solute mass / solution mass

[0053] Select liquid Pluronic F127 at a temperature of 4-5°C. When the material is sucked into the micropipette on the print head, it is immersed in P...

Embodiment 2

[0068] Cell handling:

[0069] 1) Take out the culture dish containing human neonatal dermal fibroblasts (hNDFs) from the incubator, wash with phosphate buffered saline (PBS), and digest the cells with trypsin;

[0070] 2) The treated cells were resuspended in 4 mL of cell culture medium and placed in a 10 mL tissue culture flask;

[0071] 3) The culture flask was placed on a rotary shaker at a volume concentration of 5% CO 2 Environment, culture at 37°C for 1 hour, centrifuge at 3500rpm;

[0072] 4) Use a 3D printer to extrude the cell column on a special, non-adhesive polytetrafluoroethylene or agarose base, and culture it overnight to mature the cell column;

[0073] Poly-N-isopropylacrylamide (PNIPAAm) treatment:

[0074] Select liquid poly-N-isopropylacrylamide, when the material is sucked into the micropipette on the print head, immerse in PBS to solidify the material, and continuously extruded strips of poly-N-isopropylacrylamide can be obtained;

[0075] 3D blood v...

Embodiment 3

[0088] Cell handling:

[0089] 1) Take out the culture dish containing human neonatal dermal fibroblasts (hNDFs) from the incubator, wash with phosphate buffered saline (PBS), and digest the cells with trypsin;

[0090] 2) The treated cells were resuspended in 4 mL of cell culture medium and placed in a 10 mL tissue culture flask;

[0091] 3) The culture flask was placed on a rotary shaker at a volume concentration of 5% CO 2 Environment, culture at 37°C for 1 hour, centrifuge at 3500rpm;

[0092] 4) Use a 3D printer to extrude the cell column on a special, non-adhesive polytetrafluoroethylene or agarose base, and culture it overnight to mature the cell column;

[0093] Methylcellulose (MC) treatment:

[0094] The content of methylcellulose (MC) is 12%-16%, and PBS is added to form the MC-water-salt system.

[0095] Select the above-mentioned liquid methylcellulose, when the material is sucked into the micropipette on the print head, immerse in PBS above 32°C to solidify t...

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Abstract

The invention provides a method for preparing a tissue engineering blood vessel based on a 3D (3-Dimensional) bioprinting technology. The method comprises the following steps:alternately printing a blood vessel shape by virtue of a high-polymer material and a human renascent skin fiber cell column, and removing the high-polymer material after fusion to structure a tissue engineering blood vessel; filling late-outgrowth endothelial progenitor cells into the blood vessel, and performing in-vitro dynamic culturing for 7 days in a bioreactor to obtain a differentiated tissue engineering blood vessel, wherein the high-polymer material is one of pluronic F127, poly(N-isopropylacrylamide), methyl cellulose and sodium alginate. The blood vessel obtained by the method is high in biocompatibility and higher in mechanical property, thrombus formation resistant and platelet adhesion resistance, and can be used for repairing and replacing a damaged or disease blood vessel.

Description

technical field [0001] The invention belongs to bioengineering technology, and particularly relates to tissue engineered blood vessels for treating vascular diseases, that is, 3D printing is used to prepare unstented tissue engineered blood vessels, which are applied to repair or replace damaged or diseased native blood vessels. Background technique [0002] Vascular replacement is one of the effective methods for treating vascular diseases. Commonly used replacement blood vessels are divided into autologous blood vessels, allogeneic blood vessels and tissue engineered blood vessels. The source of human autologous blood vessels is limited, and there is immune rejection of allogeneic blood vessels, so the research on the construction of tissue engineered blood vessels has become particularly important. [0003] The basic way to construct tissue-engineered blood vessels is to use natural or synthetic materials similar to blood vessels to construct scaffolds, plant seed cells ...

Claims

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Application Information

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IPC IPC(8): A61F2/06
CPCC12N5/0691C12N2502/1323C12N2513/00C12N2533/74C12N2533/76
Inventor 臧剑锋袁方裴梦婷戴祖明黄琦
Owner 江苏知聚知识产权服务有限公司
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