CN113189317A - Experimental device for three-dimensional static culture of artificial blood vessel and use method thereof - Google Patents
Experimental device for three-dimensional static culture of artificial blood vessel and use method thereof Download PDFInfo
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- CN113189317A CN113189317A CN202110473055.4A CN202110473055A CN113189317A CN 113189317 A CN113189317 A CN 113189317A CN 202110473055 A CN202110473055 A CN 202110473055A CN 113189317 A CN113189317 A CN 113189317A
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- 210000004204 blood vessel Anatomy 0.000 title claims abstract description 35
- 239000002473 artificial blood Substances 0.000 title claims abstract description 23
- 230000003068 static effect Effects 0.000 title claims abstract description 23
- 238000000034 method Methods 0.000 title claims abstract description 9
- 239000000017 hydrogel Substances 0.000 claims abstract description 43
- 238000007493 shaping process Methods 0.000 claims abstract description 14
- 239000001963 growth medium Substances 0.000 claims description 14
- 239000000463 material Substances 0.000 claims description 10
- 150000001875 compounds Chemical class 0.000 claims description 9
- 230000000149 penetrating effect Effects 0.000 claims description 5
- 239000002131 composite material Substances 0.000 claims description 4
- 239000011521 glass Substances 0.000 claims description 3
- 230000002792 vascular Effects 0.000 description 5
- 230000008929 regeneration Effects 0.000 description 3
- 238000011069 regeneration method Methods 0.000 description 3
- 208000024172 Cardiovascular disease Diseases 0.000 description 2
- 230000004071 biological effect Effects 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 238000000338 in vitro Methods 0.000 description 2
- 238000000465 moulding Methods 0.000 description 2
- 239000005020 polyethylene terephthalate Substances 0.000 description 2
- 229920004934 Dacron® Polymers 0.000 description 1
- 208000005189 Embolism Diseases 0.000 description 1
- 208000032984 Intraoperative Complications Diseases 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 229920000295 expanded polytetrafluoroethylene Polymers 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 229910010272 inorganic material Inorganic materials 0.000 description 1
- 239000011147 inorganic material Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- -1 polyethylene terephthalate Polymers 0.000 description 1
- 229920000139 polyethylene terephthalate Polymers 0.000 description 1
- 230000000250 revascularization Effects 0.000 description 1
- 238000009958 sewing Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
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- G—PHYSICS
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- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
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- G09B23/28—Models for scientific, medical, or mathematical purposes, e.g. full-sized devices for demonstration purposes for medicine
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/36—Embedding or analogous mounting of samples
- G01N2001/366—Moulds; Demoulding
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Abstract
The invention discloses a set of experimental device for three-dimensional static culture of an artificial blood vessel and a using method thereof, wherein the experimental device comprises a culture tank, a culture tube, a culture chamber and a hydrogel shaping mold, wherein the culture tank is cuboid and is not covered; the wide side of the culture tank is downwards sunken into an arc shape; two wide edges of the culture tank are correspondingly provided with semi-arc-shaped culture tube supporting grooves; the culture chamber comprises two hollow hemispheroids; the two hollow hemispheroids are fixedly arranged together; the surface of the culture chamber is provided with a plurality of holes; two culture room supporting grooves penetrate through the joint of the two hollow hemispheroids along the diameter direction; the hydrogel shaping mold comprises a left clamping plate, a right clamping plate and a base; the base is arranged below the left clamping plate and embedded into the right clamping plate, and semi-cylindrical holes are symmetrically formed above the left clamping plate and the right clamping plate; the semi-cylindrical holes are combined to form a cylindrical hole; the culture tube is cylindrical, and the size of the culture tube is matched with the culture tube supporting groove, the culture chamber supporting groove and the cylindrical hole. The invention can better simulate the three-dimensional growth of human blood vessels in natural conditions.
Description
Technical Field
The invention relates to the technical field of medical treatment, in particular to a set of experimental device for three-dimensional static culture of an artificial blood vessel and a using method thereof.
Background
In China, 300-400 million people still die each year due to cardiovascular diseases, and the vascular reconstruction plays an important role in the treatment process of the cardiovascular diseases. The main mode of vascular building is autologous arterial or venous vascular grafting. However, autologous vascular grafts present difficulties and challenges for current revascularization conditions due to limitations and secondary surgical injury. According to study statistics, more than 10% of patients cannot be transplanted with autologous blood vessels. Later researchers began focusing on the development of vascular substitutes such as expanded polytetrafluoroethylene-ePTFE and polyethylene terephthalate fiber-dacron, among others. However, such foreign substances have a series of problems in vivo such as occurrence of embolism, lack of regeneration function, and the like. Therefore, the regeneration and cultivation of the in vitro tissue engineering blood vessel provides possibility for new blood vessel construction.
The regeneration culture of the in vitro tissue engineering blood vessel mainly comprises three parts: seed cells, a culture bracket and a culture environment. The culture scaffold has various choices, and among them, PGA inorganic materials and hydrogels are used in many cases. There are many culture models for PGA materials, but there are few hydrogel-type experimental devices, and many experiments are simulated in a two-dimensional environment, which is greatly different from the original blood vessel growth environment and cannot well simulate the three-dimensional growth of blood vessels.
Disclosure of Invention
The invention aims to overcome the defects and shortcomings of the prior art, provides a set of experimental device for artificial blood vessel three-dimensional static culture, and can better simulate the three-dimensional growth of human blood vessels in natural conditions.
The invention also aims to provide a use method of the experimental device for the three-dimensional static culture of the artificial blood vessel.
The purpose of the invention can be realized by the following technical scheme: a set of experimental device for three-dimensional static culture of artificial blood vessels comprises a culture tank, a culture tube, a culture chamber and a hydrogel shaping mold;
the culture tank is in a cuboid uncovered shape; the wide side of the culture tank is downwards sunken into an arc shape; and semicircular arc-shaped culture tube supporting grooves are correspondingly formed in the two wide edges of the culture tank.
The culture chamber comprises two hollow hemispheroids; the two hollow hemispheroids are fixedly arranged together; the surface of the culture chamber is provided with a plurality of holes; two culture room supporting grooves penetrate through the joint of the two hollow hemispheroids along the diameter direction;
the hydrogel shaping mold comprises a left clamping plate, a right clamping plate and a base; the base is arranged below the left clamping plate and embedded into the right clamping plate, and semi-cylindrical holes are symmetrically formed above the left clamping plate and the right clamping plate; the semi-cylindrical holes are combined to form a cylindrical hole;
the culture tube is cylindrical, and the size of the culture tube is matched with that of the culture tube support groove, the culture chamber support groove and the cylindrical hole.
Furthermore, two strip-shaped fixing grooves are symmetrically formed in the two hollow hemispheroids respectively. The two hollow hemispheroids can be fixedly combined by a rope which correspondingly penetrates through the fixing groove.
Further, the semi-cylindrical holes are combined to form two cylindrical holes; and a screw hole is formed from the side surface of the left clamping plate to the right clamping plate in a penetrating way. The left clamping plate and the right clamping plate are fixedly combined together through screws.
Furthermore, the culture tank is made of ABS material. ABS is an inert material and has no obvious influence on the biological activity and the growth state of cells.
Further, the culture tank has a length of 60mm, a width of 40mm and a height of 20 mm.
Furthermore, the culture tube is made of glass.
Further, the culture tube had a diameter of 6mm and a length of 30 mm.
Furthermore, the culture chamber and the hydrogel shaping mold are made of PC materials.
Further, the diameter of the culture chamber is 30 mm; the diameter of the hole in the culture chamber was 3 mm.
The other purpose of the invention can be realized by the following technical scheme: an experimental device using method for artificial blood vessel three-dimensional static culture comprises the following steps:
for the hydrogel which is soft and is not easy to shape, the hydrogel is mixed with the seed cells; spreading in a culture tank; suturing the cell hydrogel composite to the culture tube; installing the culture tube on the culture tube supporting groove or the culture chamber supporting groove; putting the culture chamber into a culture medium or injecting the culture medium into a culture tank for culture;
for the hydrogel which is hard and easy to shape, the hydrogel is mixed with the seed cells; inserting the culture tube into the cylindrical hole; injecting the cell hydrogel composite into the cylindrical hole; and demolding after the cell hydrogel compound is formed, mounting the culture tube and the cell hydrogel compound attached to the culture tube on a culture tube supporting groove or a culture chamber supporting groove, and putting the culture chamber into a culture medium or injecting the culture medium into the culture tank for culture.
Compared with the prior art, the invention has the following advantages and beneficial effects:
(1) the invention can obtain the cell hydrogel sheet through the culture tank, thereby being sewn into a cylindrical cell sheet which can be placed on the culture tube, the culture tube can be placed in the culture tank or the culture chamber for culture, the culture tank and the culture chamber can realize three-dimensional culture of blood vessels, and the random placement of the culture chamber further ensures that the blood vessel culture has various different culture directions, better simulates the natural conditions of human blood vessels, and thus, the invention can better simulate the three-dimensional growth of the human blood vessels.
(2) The number of culture mediums required by the whole culture is small, the whole operation is simple, and the culture cost and the culture difficulty are greatly reduced.
Drawings
FIG. 1 is a schematic view of a three-dimensional structure of a culture tank of an experimental device for three-dimensional static culture of an artificial blood vessel according to the present invention;
FIG. 2 is a schematic view of a three-dimensional structure of a culture tube of an experimental device for three-dimensional static culture of an artificial blood vessel according to the present invention;
FIG. 3 is a schematic diagram showing the three-dimensional structure of the culture chamber of the experimental apparatus for three-dimensional static culture of artificial blood vessels according to the present invention;
FIG. 4 is a schematic diagram showing a three-dimensional structure of a hydrogel molding mold of an experimental device for three-dimensional static culture of an artificial blood vessel according to the present invention.
Wherein: 1: culture tank, 11: culture tube support tank, 2: culture tube, 3: culture chamber, 31: fixing groove, 32: culture room support tank, 4: hydrogel molding die, 41: left splint, 42: right splint, 43: base, 44: cylindrical hole, 45: and a screw hole.
Detailed Description
The present invention will be described in further detail with reference to examples and drawings, but the present invention is not limited thereto.
A set of experimental device for three-dimensional static culture of artificial blood vessels comprises a culture tank 1, a culture tube 2, a culture chamber 3 and a hydrogel shaping mold 4.
As shown in FIG. 1, the culture tank 1 is made of ABS material, an inert material, and has no significant influence on the biological activity and growth state of cells. The whole body is in a cuboid uncovered shape, the length is 60mm, the width is 40mm, the height is 20mm, and the wide edge is downwards sunken into an arc shape. The middle parts of the two wide edges are respectively provided with a culture tube supporting groove 11 with a diameter of 6mm in a semicircular arc shape and can be used for placing the culture tube 2.
As shown in FIG. 2, the culture tube 2 is a cylindrical glass column having a diameter of 6mm and a length of 30 mm.
As shown in FIG. 3, the culture chamber 3 is made of PC material, and comprises two hollow hemispheroids with 30mm diameter, two elongated fixing grooves 31 are symmetrically formed in the two hollow hemispheroids respectively, a rope can correspondingly penetrate through the fixing grooves 31 to fixedly combine the two hollow hemispheroids, a plurality of holes with 3mm diameter are formed in the surface of the hemispheroids, and when the culture chamber is placed in a culture medium, the culture medium can flow into the culture chamber through the holes. Two culture room supporting grooves 32 are formed in the joint of the two hollow hemispheroids in the diameter direction of the culture room 3 in a penetrating manner, and the culture tube 2 can pass through the two culture room supporting grooves 32 and be installed on the culture room 3.
As shown in fig. 4, the hydrogel shaping mold 4 is made of PC material, is rectangular, has a length of 80mm, a width of 30mm and a height of 70mm, and comprises a left clamp plate 41, a right clamp plate 42 and a base 43, wherein the base is installed below the left clamp plate 41 and embedded into the right clamp plate 42, a screw hole 45 is formed from the side surface of the left clamp plate 41 to the right clamp plate 42 in a penetrating manner, the left clamp plate and the right clamp plate are fixedly combined together through screws, and the demolding can be performed after the screws are disassembled. The upper parts of the left clamping plate 41 and the right clamping plate 42 are symmetrically provided with semi-cylindrical holes, the semi-cylindrical holes are combined to form two cylindrical holes 44, the culture tube 2 can be inserted into the two cylindrical holes 44, then the compound of the hydrogel and the seed cells is injected, after the forming, a hydrogel shaping mold is opened, the compound of the hydrogel and the seed cells is demolded, and a tubular cell hydrogel compound with the inner diameter of 6mm, the outer diameter of 10mm and the length of 30mm can be cast.
The use method of the experimental device comprises the following steps:
for soft hydrogel which is not easy to shape, firstly mixing seed cells with the hydrogel in a culture tank, performing flat culture until the seed cells and the hydrogel are combined into a sheet, then manually sewing the sheet to a culture tube, putting a culture tube frame in a culture chamber support groove of a culture chamber, and finally putting the culture chamber into a culture medium for culture; or the culture medium is injected into the culture tank, and then the culture tube is placed on the culture tube support groove of the culture tank to culture.
For the hydrogel which is hard and easy to shape, mixing seed cells with the hydrogel, inserting a culture tube into a cylindrical hole of a hydrogel shaping die, then injecting a cell hydrogel compound, opening the hydrogel shaping die after shaping, taking out the culture tube and the cell hydrogel compound attached to the culture tube, placing the culture tube frame in a culture chamber supporting groove of a culture chamber, and finally placing the culture chamber in a culture medium for culture; or the culture medium is injected into the culture tank, and then the culture tube is placed on the culture tube support groove of the culture tank to culture.
The above-mentioned embodiments only express several embodiments of the present invention, and the description thereof is more specific and detailed, but not construed as limiting the scope of the present invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention. Therefore, the protection scope of the present patent shall be subject to the appended claims.
Claims (10)
1. A set of experimental device for three-dimensional static culture of artificial blood vessels is characterized by comprising a culture tank (1), a culture tube (2), a culture chamber (3) and a hydrogel shaping mold (4);
the culture tank (1) is in a cuboid uncovered shape; the wide side of the culture tank (1) is downwards sunken into an arc shape; two wide edges of the culture tank (1) are correspondingly provided with semi-circular arc-shaped culture tube supporting grooves (11);
the culture chamber (3) comprises two hollow hemispheroids; the two hollow hemispheroids are fixedly arranged together; a plurality of holes are formed in the surface of the culture chamber (3); two culture room supporting grooves (32) are formed at the joint of the two hollow hemispheroids in a penetrating manner along the diameter direction;
the hydrogel shaping mold (4) comprises a left clamping plate (41), a right clamping plate (42) and a base (43); the base (43) is arranged below the left clamping plate (41) and embedded into the right clamping plate (42), and semi-cylindrical holes are symmetrically formed above the left clamping plate (41) and the right clamping plate (42); the semi-cylindrical holes combine to form a cylindrical hole (44);
the culture tube (2) is cylindrical, and the size of the culture tube is matched with the culture tube supporting groove (11), the culture chamber supporting groove (32) and the cylindrical hole (44).
2. The experimental device for three-dimensional static culture of artificial blood vessels as claimed in claim 1, wherein two strip-shaped fixing grooves (31) are symmetrically formed on the two hollow hemispheroids respectively.
3. The set of experimental devices for three-dimensional static culture of artificial blood vessels according to claim 1, wherein the semi-cylindrical holes are combined to form two cylindrical holes (44); a screw hole (45) is formed from the side surface of the left clamping plate (41) to the right clamping plate (42) in a penetrating manner.
4. The set of experimental device for three-dimensional static culture of artificial blood vessels according to claim 1, wherein the culture tank (1) is made of ABS material.
5. The set of experimental devices for three-dimensional static culture of artificial blood vessels according to claim 1, wherein the culture tank (1) has a length of 60mm, a width of 40mm and a height of 20 mm.
6. The experimental device for three-dimensional static culture of artificial blood vessels according to claim 1, wherein the culture tube (2) is made of glass.
7. The set of experimental devices for three-dimensional static culture of artificial blood vessels according to claim 1, wherein the culture tube (2) has a diameter of 6mm and a length of 30 mm.
8. The set of experimental apparatus for three-dimensional static culture of artificial blood vessel according to claim 1, wherein the culture chamber (3) and the hydrogel shaping mold (4) are made of PC material.
9. The set of experimental devices for the three-dimensional static culture of artificial blood vessels according to claim 1, characterized in that the culture chamber (3) has a diameter of 30 mm; the diameter of the hole on the culture chamber (3) is 3 mm.
10. The use method of the set of experimental devices for three-dimensional static culture of artificial blood vessels as claimed in any one of claims 1 to 9, which is characterized by comprising the following steps:
for the hydrogel which is soft and is not easy to shape, the hydrogel is mixed with the seed cells; spread in the culture tank (1); suturing the cell hydrogel composite to the culture tube (2); installing the culture tube (2) on the culture tube support tank (11) or the culture chamber support tank (32); putting the culture chamber into a culture medium or injecting the culture medium into a culture tank for culture;
for the hydrogel which is hard and easy to shape, the hydrogel is mixed with the seed cells; inserting the culture tube (2) into the cylindrical hole (44); injecting a cell hydrogel composite into the cylindrical bore (44); and (3) demolding after the cell hydrogel compound is formed, mounting the culture tube (2) and the cell hydrogel compound attached to the culture tube (2) on a culture tube supporting groove (11) or a culture chamber supporting groove (32), and putting the culture chamber into a culture medium or injecting the culture medium into the culture tank for culture.
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| CN202110473055.4A CN113189317A (en) | 2021-04-29 | 2021-04-29 | Experimental device for three-dimensional static culture of artificial blood vessel and use method thereof |
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