Blood vessel vitrifying storage liquid and blood vessel storage method

Abstract

The present invention relates to blood vessel storing liquid for fast vitrifying and cryogenically storing body blood vessel, artificial blood vessel and tissue engineering blood vessel and the corresponding storage method. The blood vessel vitrifying and storing liquid is compounded through diluting polysaccharose and mycose with M199 diluent to obtain mixed solution of 20-35 % and 15-25 % concentration separately; diluting vinyl glycol and vinyl alcohol with the mixed solution to mixed liquid of 20 % concentration; and adding azone as high penetrant in 0.5-1 % to form the blood vessel vitrifying and storing liquid, into which, 0.001-0.005 % of cationic surfactant may be also added. Blood vessel is thrown into the vitrifying and storing liquid inside some plastic bag at room temperature, and the sealed bag is thrown into liquid nitrogen for cryogenical storing. The cryogenically stored blood vessel may be defrosted via taking out the bag from liquid nitrogen and soaking in water bath at 22-25 deg.c for 15 sec.

Description

technical field

[0001] The invention belongs to the technical field of preservation of human or animal bodies or their parts or tissue engineering products, and particularly relates to a blood vessel preservation solution and a preservation method for rapid vitrification deep-low temperature human blood vessels, artificial blood vessels, and tissue engineering blood vessels. technical background:

[0002] At present, the incidence of cardiovascular disease has become one of the main causes of human death. In the United States alone, 600,000 people need coronary artery bypass graft surgery every year. In my country, according to the latest statistics, elderly people over 60 years old In 2000, it reached 130 million, ranking first in the world. Vascular bypass and repair of vascular injury caused by other reasons all require vascular substitutes, while tissue engineered blood vessels, artificial blood vessels planted with cells, and homologous blood vessels must be well solved ...

Examples

Embodiment 1

[0022] Example 1: Dilute Ficoll70 (the average relative molecular mass is 70,000) and trehalose into 30% and 17% mixed solutions (FT solution) with M199 solution, add 1% azone, and then dilute vinylglycerin with FT solution Alcohol and polyvinyl alcohol (20:1) are mixed and diluted to 30% (v / v) PEFT30 vitrification solution, and 0.002% of anionic surfactants such as sodium lauryl sulfate are added. At a room temperature of 20°C, transfer the constructed tissue engineered blood vessel or artificial blood vessel into a 50ml plastic tube or plastic bag containing PEFT30 vitrification solution to equilibrate for 1 minute, seal it and put it directly into liquid nitrogen for cryopreservation. When thawing is required, take the cryopreserved tube or bag out of the liquid nitrogen and immerse it directly in a 25°C water bath for about 20 seconds to complete the thawing procedure.

[0023] The above-mentioned thawed blood vessels were digested with enzymatic digestion method to digest...

Embodiment 2

[0024] Embodiment 2: Ficoll70 (average relative molecular mass is 70000) and trehalose (Trehalose) are diluted into 30% and 17% mixed solution (FT solution) with M199 liquid, then ethylene glycol and polyvinyl alcohol are respectively mixed with FT solution (30:1) mixed and diluted to 40% (v / v) PEFT40 vitrification solution, adding 0.8% azone and a small amount of anionic surfactants such as sodium lauryl sulfate. At a room temperature of 25°C, transfer the aseptically obtained homologous or heterologous blood vessels into a 50ml plastic tube or plastic bag containing PEFT40 vitrification solution to equilibrate for 1 minute, seal it and put it directly into liquid nitrogen for cryopreservation. When thawing is required, take the cryopreserved tube or bag out of the liquid nitrogen and immerse it directly in a 25°C water bath for about 20 seconds to complete the thawing procedure.

[0025]The above-mentioned thawed blood vessels were digested with enzyme digestion method to di...

technical field

[0001] The invention belongs to the technical field of preservation of human or animal bodies or their parts or tissue engineering products, and particularly relates to a blood vessel preservation solution and a preservation method for rapid vitrification deep-low temperature human blood vessels, artificial blood vessels, and tissue engineering blood vessels. technical background:

[0002] At present, the incidence of cardiovascular disease has become one of the main causes of human death. In the United States alone, 600,000 people need coronary artery bypass graft surgery every year. In my country, according to the latest statistics, elderly people over 60 years old In 2000, it reached 130 million, ranking first in the world. Vascular bypass and repair of vascular injury caused by other reasons all require vascular substitutes, while tissue engineered blood vessels, artificial blood vessels planted with cells, and homologous blood vessels must be well solved ...

Examples

Embodiment 1

[0022] Example 1: Dilute Ficoll70 (the average relative molecular mass is 70,000) and trehalose into 30% and 17% mixed solutions (FT solution) with M199 solution, add 1% azone, and then dilute vinylglycerin with FT solution Alcohol and polyvinyl alcohol (20:1) are mixed and diluted to 30% (v / v) PEFT30 vitrification solution, and 0.002% of anionic surfactants such as sodium lauryl sulfate are added. At a room temperature of 20°C, transfer the constructed tissue engineered blood vessel or artificial blood vessel into a 50ml plastic tube or plastic bag containing PEFT30 vitrification solution to equilibrate for 1 minute, seal it and put it directly into liquid nitrogen for cryopreservation. When thawing is required, take the cryopreserved tube or bag out of the liquid nitrogen and immerse it directly in a 25°C water bath for about 20 seconds to complete the thawing procedure.

[0023] The above-mentioned thawed blood vessels were digested with enzymatic digestion method to digest...

Embodiment 2

[0024] Embodiment 2: Ficoll70 (average relative molecular mass is 70000) and trehalose (Trehalose) are diluted into 30% and 17% mixed solution (FT solution) with M199 liquid, then ethylene glycol and polyvinyl alcohol are respectively mixed with FT solution (30:1) mixed and diluted to 40% (v / v) PEFT40 vitrification solution, adding 0.8% azone and a small amount of anionic surfactants such as sodium lauryl sulfate. At a room temperature of 25°C, transfer the aseptically obtained homologous or heterologous blood vessels into a 50ml plastic tube or plastic bag containing PEFT40 vitrification solution to equilibrate for 1 minute, seal it and put it directly into liquid nitrogen for cryopreservation. When thawing is required, take the cryopreserved tube or bag out of the liquid nitrogen and immerse it directly in a 25°C water bath for about 20 seconds to complete the thawing procedure.

[0025]The above-mentioned thawed blood vessels were digested with enzyme digestion method to di...