Kit for obtaining multiple-organ and tissue extracellular matrix and using method of kit

A technology for tissue cells and extracellular matrix, applied in medical science, prostheses, etc., can solve problems such as narrow application range, large amount of DNA residue, and narrow range

Active Publication Date: 2015-01-21
ZHEJIANG DISAI BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the preparation methods of decellularized scaffolds for each whole organ or tissue are complicated and diverse, and the reagents used are also relatively messy, lacking systematic arrangement
[0003] Cheng Yuan and others disclosed a kit for preparing decellularized liver and its use method, although the above kit can be used to perfuse the liver sequentially with thrombolytic solution, perfusion solution I, II, III, and IV to produce decellularized liver Scaffold, but its kit can only be used to prepare decellularized scaffolds for the liver of some animals, and the steps are relatively cumbersome, and because the method and reagents are relatively simpl

Method used

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  • Kit for obtaining multiple-organ and tissue extracellular matrix and using method of kit
  • Kit for obtaining multiple-organ and tissue extracellular matrix and using method of kit
  • Kit for obtaining multiple-organ and tissue extracellular matrix and using method of kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Take the non-ionic detergent, hypotonic solution, isotonic solution, tissue loosening agent and nucleic acid remover with a solute-to-solvent volume ratio of 0.1% in the kit for obtaining the extracellular matrix of multiple organs and tissues.

[0040] In vitro, the heart was flushed out of residual blood through the ascending aorta with embolism solution, and placed in a low-temperature refrigerator at -80°C for 12 to 4 hours. Each time lasted 20 minutes, perfused tissue loosening agent with hypotonic solution for 15 minutes, perfused tissue loosening agent with non-ionic detergent for 6 hours, treated with nuclease remover for 30 minutes, rinsed with isotonic solution for 2 hours, and perfused at a rate of 1ml / min. All were carried out continuously; the prepared porcine heart cell scaffold, the appearance of the cardiac decellularized scaffold, the transparent capsule completely retained the shape of the heart, and the vasculature in the heart was visible to the naked...

Embodiment 2

[0042] Take the non-ionic detergent, hypotonic solution, isotonic solution, tissue loosening agent and nucleic acid remover in the kit for obtaining the extracellular matrix of multiple organs and tissues with a volume ratio of solute to solvent of 3%.

[0043] The heart was flushed out of residual blood through the ascending aorta in vitro, placed in a low-temperature refrigerator at -40°C for 30 hours, and then thawed in isotonic sterile PBS buffer solution at 30°C after 30 hours, rinsed three times with normal saline, Each time lasted 20 minutes, infused with tissue loosening agent in hypotonic solution for 40 minutes, infused with tissue loosening agent in non-ionic detergent for 15 hours, treated with nuclease remover for 45 minutes, rinsed with isotonic solution for 15 hours, and the perfusion speed was 200ml / min. The steps are all carried out continuously; the prepared porcine heart cell scaffold, the appearance of the cardiac decellularized scaffold, the transparent cap...

Embodiment 3

[0045] Take the non-ionic detergent, hypotonic solution, isotonic solution, tissue loosening agent and nucleic acid remover in the kit for obtaining the extracellular matrix of multiple organs and tissues with a volume ratio of 10% solute to solvent.

[0046] In vitro, the heart was flushed out of the residual blood through the ascending aorta with embolism solution, placed in a low-temperature refrigerator at -20°C for 48h12-48h, and then thawed in isotonic sterile PBS buffer solution at 45°C, and wasted with isotonic solution or normal saline Rinse 3 times, each lasting 20 minutes, perfuse tissue loosening agent with hypotonic solution for 60 minutes, infuse tissue loosening agent with non-ionic detergent for 24 hours, treat with nuclease remover for 60 minutes, rinse with isotonic solution for 24 hours, and infuse at a rate of 500ml / min, all steps are carried out continuously; the prepared porcine heart cell scaffold, the appearance of the cardiac decellularized scaffold, t...

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Abstract

The invention discloses a kit for obtaining a multiple-organ and tissue extracellular matrix and a using method of the kit. The kit for obtaining the multiple-organ and tissue extracellular matrix comprises a nonionic detergent I, an ionic detergent I, an ionic detergent II, a hypertonic solution, a hypotonic solution, an isotonic solution, a nucleic acid removal agent and a tissue bulking agent. According to the kit and a method for using the kit provided by the invention, acellularized scaffolds of multiple organs and tissues can be obtained in a short time, and the structure and most of structure of the extracellular matrix are maintained, so that product materials are provided for research and application of regenerative medicines and tissue engineering.

Description

technical field [0001] The invention relates to a kit for obtaining extracellular matrices of multiple organs and tissues, in particular to a kit for removing antigenic components of different organs and tissues. The invention also contemplates methods for obtaining multiple organ and tissue scaffolds using the kit. Background technique [0002] In recent years, decellularized tissue engineering has developed rapidly. At present, decellularized scaffolds from various tissue sources have been successfully used in tissue engineering and regenerative medicine, such as skin, blood vessels, nerves, small intestinal submucosa, and liver. In addition to the more mature decellularized tissues mentioned above, decellularized scaffolds of whole organs of many animals have also been successfully prepared, such as heart, liver, kidney, lung, spinal cord, etc., which laid a solid foundation for allogeneic transplantation under the current situation of tight demand for tissue and organ tr...

Claims

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Application Information

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IPC IPC(8): A61L27/36
Inventor 倪金虎林贤丰张琪何佳怡陈蕴缤陈家鑫
Owner ZHEJIANG DISAI BIOTECHNOLOGY CO LTD
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