CN115247126A - Artificial blood vessel extracorporeal circulation culture system and use method thereof - Google Patents
Artificial blood vessel extracorporeal circulation culture system and use method thereof Download PDFInfo
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- CN115247126A CN115247126A CN202111653486.5A CN202111653486A CN115247126A CN 115247126 A CN115247126 A CN 115247126A CN 202111653486 A CN202111653486 A CN 202111653486A CN 115247126 A CN115247126 A CN 115247126A
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- 210000004204 blood vessel Anatomy 0.000 title claims abstract description 99
- 239000002473 artificial blood Substances 0.000 title claims abstract description 93
- 230000004087 circulation Effects 0.000 title claims abstract description 23
- 238000000034 method Methods 0.000 title claims abstract description 17
- 230000002572 peristaltic effect Effects 0.000 claims abstract description 30
- 239000001963 growth medium Substances 0.000 claims abstract description 29
- 239000007788 liquid Substances 0.000 claims abstract description 28
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 22
- 238000004113 cell culture Methods 0.000 claims description 19
- 230000035755 proliferation Effects 0.000 claims description 7
- 238000009530 blood pressure measurement Methods 0.000 claims description 4
- 230000003068 static effect Effects 0.000 claims description 4
- 239000011521 glass Substances 0.000 claims description 3
- 229920001296 polysiloxane Polymers 0.000 claims description 3
- 230000017531 blood circulation Effects 0.000 abstract description 6
- 230000021164 cell adhesion Effects 0.000 abstract description 6
- 238000005086 pumping Methods 0.000 abstract description 4
- 238000012360 testing method Methods 0.000 abstract description 4
- 238000002474 experimental method Methods 0.000 abstract description 3
- 238000000338 in vitro Methods 0.000 description 7
- 238000011156 evaluation Methods 0.000 description 6
- 238000001514 detection method Methods 0.000 description 5
- 230000008901 benefit Effects 0.000 description 3
- 238000001727 in vivo Methods 0.000 description 3
- 230000000694 effects Effects 0.000 description 2
- 230000006978 adaptation Effects 0.000 description 1
- 230000036772 blood pressure Effects 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 230000003013 cytotoxicity Effects 0.000 description 1
- 231100000135 cytotoxicity Toxicity 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
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- C12M21/00—Bioreactors or fermenters specially adapted for specific uses
- C12M21/08—Bioreactors or fermenters specially adapted for specific uses for producing artificial tissue or for ex-vivo cultivation of tissue
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- C12M23/00—Constructional details, e.g. recesses, hinges
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- C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
- C12M41/30—Means for regulation, monitoring, measurement or control, e.g. flow regulation of concentration
- C12M41/32—Means for regulation, monitoring, measurement or control, e.g. flow regulation of concentration of substances in solution
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- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/06—Animal cells or tissues; Human cells or tissues
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Abstract
The invention discloses an artificial blood vessel extracorporeal circulation culture system and a use method thereof, and the system comprises a frequency converter, an artificial blood vessel, a pressure measurer, a culture medium bottle, a peristaltic pump and a liquid collecting box, wherein the pipelines comprise a first pipeline, a second pipeline, a third pipeline and a fourth pipeline, the artificial blood vessel is arranged in the liquid collecting box, two ends of the artificial blood vessel are respectively communicated with one ends of the first pipeline and the third pipeline, one end of the peristaltic pump is communicated with the first pipeline, the other end of the peristaltic pump is communicated with one end of the second pipeline, the other end of the second pipeline is communicated with a culture medium, the other end of the third pipeline is communicated with the culture medium bottle, one end of the fourth pipeline is communicated with the culture medium bottle, the other end of the fourth pipeline is communicated with the liquid collecting box, a water stopping clamp is arranged on the fourth pipeline, the pressure measurer is arranged on the first pipeline, and the frequency converter is used for controlling the water pumping quantity of the peristaltic pump; the system can simulate the cell adhesion condition of the artificial blood vessel under different pressures and blood flow environments, and can simultaneously test a plurality of artificial blood vessels to improve the efficiency and the repeatability of the experiment.
Description
Technical Field
The invention relates to the technical field of medical instrument testing devices, in particular to an artificial blood vessel extracorporeal circulation culture system and a use method thereof.
Background
The artificial blood vessel has good social benefit and considerable economic benefit when being applied to clinic. The artificial blood vessel must be evaluated for biocompatibility during the development process and the biocompatibility must meet the clinical requirements so as to be applicable to the clinic. If biocompatibility does not meet clinical requirements, the artificial blood vessel must be modified to improve its biocompatibility.
At present, the evaluation of biocompatibility of an artificial blood vessel requires the evaluation of cellular compatibility, hemocompatibility and histocompatibility as specified in international standard ISO 10993. The evaluation of cell compatibility requires the evaluation of cytotoxicity under static conditions, and the evaluation of histocompatibility requires in vivo animal experiments, which often result in experimental results that do not match the results obtained under static conditions. If the biocompatibility of the artificial blood vessel can be evaluated by simulating the dynamic environment in vivo in vitro, a more objective experimental result can be obtained.
However, the blood pressure and the blood flow rate are different at different parts of the human body, and thus, the pressure applied to the inner wall of the artificial blood vessel transplanted to different parts is also different. When in vitro cell compatibility evaluation is carried out, the blood flow effect on blood vessel cells at different parts in vivo is difficult to simulate, so that the design of an artificial blood vessel in vitro circulation culture system aiming at the pressure of a transplanted part is particularly urgent. Through retrieval, no related experimental device or instrument exists at home and abroad at present, and the dynamic culture cell compatibility of the artificial blood vessel material can be tested under the conditions of controllable pressure and flow rate.
In order to solve the above problems, the present invention provides an artificial blood vessel extracorporeal circulation culture system and a use method thereof, so as to solve the problem that the existing detection device cannot complete the cell compatibility detection of the artificial blood vessel under different pressures and different blood flow rates.
Disclosure of Invention
The invention aims to provide an artificial blood vessel extracorporeal circulation culture system and a use method thereof, which achieve the purpose of detecting the cell compatibility of the artificial blood vessel under different pressures and different blood flow rates.
In order to achieve the purpose, the invention provides the following scheme:
the utility model provides an artificial blood vessel extracorporeal circulation culture system, includes converter, artificial blood vessel, pressure measurement ware, communicates in proper order and forms closed loop's culture medium bottle, peristaltic pump and collection liquid case through the pipeline, the pipeline includes first pipeline, second pipeline, third pipeline and fourth pipeline, artificial blood vessel sets up in the collection liquid case and both ends communicate with the one end of first pipeline and third pipeline respectively, the one end of peristaltic pump with first pipeline intercommunication, the other end with the one end intercommunication of second pipeline, the other end of second pipeline with the culture medium intercommunication, the other end of third pipeline with culture medium bottle intercommunication, fourth pipeline one end with culture medium bottle intercommunication, the other end with the collection liquid case intercommunication, install the stagnant water clamp on the fourth pipeline, the pressure measurement ware sets up on the first pipeline, the converter is used for controlling the pump water yield of pump.
Preferably, a first joint is arranged at the position where the first pipeline is communicated with the liquid collecting tank, and a second joint is arranged at the position where the third pipeline is communicated with the liquid collecting tank.
Preferably, the first joint and the second joint are both straight joints or forked multi-way joints.
Preferably, the first pipeline, the second pipeline, the third pipeline and the fourth pipeline are all silicone tubes without biotoxicity.
Preferably, the liquid collection tank and the culture medium bottle are transparent glass water storage containers.
Preferably, the pressure detector comprises a pressure sensor and a pressure gauge, the pressure sensor is arranged on the first pipeline and close to the first joint, the pressure sensor is connected with the pressure gauge, and the pressure gauge is used for displaying the pressure in the artificial blood vessel.
The use method of the artificial blood vessel extracorporeal circulation culture system comprises the following steps:
step 1: injecting complete cell culture solution into the culture medium bottle, respectively connecting two ends of an artificial blood vessel cultured together with the cell culture solution with the first joint and the second joint, opening the peristaltic pump, controlling the flow of the peristaltic pump through the frequency converter, and enabling the pressure in the artificial blood vessel and the flow rate of the cell culture solution to reach preset values;
step 2: opening the water stop clamp, and enabling complete cell culture solution seeped out of the artificial blood vessel to enter the culture medium bottle through a fourth pipeline;
and 3, step 3: keeping the artificial blood vessel extracorporeal circulation culture system running for a preset time, closing the frequency converter and the peristaltic pump, taking out the blood vessel sample, and observing the adhesion and proliferation conditions of cells on the inner wall of the artificial blood vessel.
Preferably, the culture process of the artificial blood vessel in step 1 is as follows: and closing the water stop clamp, respectively connecting the two ends of the artificial blood vessel with the first connector and the second connector, injecting complete cell culture solution into the liquid collecting box, immersing the artificial blood vessel, and performing static culture for 4 to 5 hours.
Preferably, the instruments used for observing the adhesion and proliferation of cells on the inner wall of the artificial blood vessel in the step 3 are a scanning electron microscope and a fluorescence microscope.
Preferably, the pressure value of the artificial blood vessel in the step 1 changes within the range of +/-0.3 kPa in continuous time, namely, the stable pressure is reached.
Compared with the prior art, the invention has the following technical effects:
1. according to the invention, different water pumping amounts of the peristaltic pump are controlled by the frequency converter, so that the artificial blood vessel generates different pressure values and different liquid flow rate values, and the cell compatibility under different pressures and different flow rates is further detected; the dynamic setting mode of the whole circulatory system can test the cell adhesion condition of the artificial blood vessel under the dynamic condition; meanwhile, the arrangement mode of the first joint and the second joint is a forked multi-way type, so that a plurality of artificial blood vessels can be tested at the same time to improve the efficiency and the repeatability of the experiment.
Drawings
In order to more clearly illustrate the present invention or technical solutions in the prior art, the drawings needed to be used in the embodiments will be briefly described below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and it is obvious for those skilled in the art to obtain other drawings without inventive exercise.
FIG. 1 is a schematic structure of an artificial blood vessel extracorporeal circulation culture system;
FIG. 2 is a schematic diagram of a bifurcated artificial blood vessel;
FIG. 3 is a schematic view of a plurality of artificial blood vessels;
wherein, 1, a culture medium bottle; 2. a second joint; 3. a liquid collection tank; 4. a liquid discharge port; 5. a water stop clip; 6. an artificial blood vessel; 7. a first joint; 8. a pressure gauge; 9. a pressure sensor; 10. a peristaltic pump; 11. a frequency converter; 12. a fourth pipeline; 13. a second pipeline; 14. a first pipeline; 15. a third pipeline.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be obtained by a person skilled in the art without making any creative effort based on the embodiments in the present invention, belong to the protection scope of the present invention.
The invention aims to provide an artificial blood vessel in-vitro circulation culture system and a using method thereof, which achieve the purpose of detecting the cell compatibility of the artificial blood vessel under different pressures and different blood flow rates.
In order to make the aforementioned objects, features and advantages of the present invention comprehensible, embodiments accompanied with figures are described in further detail below.
Referring to fig. 1, an artificial blood vessel extracorporeal circulation culture system comprises a frequency converter 11, an artificial blood vessel 6, a pressure measurer, a culture medium bottle 1, a peristaltic pump 10 and a liquid collection tank 3 which are sequentially communicated through a pipeline and form a closed loop, wherein the pipeline comprises a first pipeline 14, a second pipeline 13, a third pipeline 15 and a fourth pipeline 12, the artificial blood vessel 6 is arranged in the liquid collection tank 3, two ends of the artificial blood vessel are respectively communicated with one ends of the first pipeline 14 and the third pipeline, one end of the peristaltic pump 10 is communicated with the first pipeline 14, the other end of the peristaltic pump is communicated with one end of the second pipeline 13, the other end of the second pipeline is communicated with a culture medium, the other end of the third pipeline 15 is communicated with the culture medium bottle 1, one end of the fourth pipeline 12 is communicated with the culture medium bottle 1, the other end of the fourth pipeline is communicated with the liquid collection tank 3, a peristaltic clamp 5 is arranged on the fourth pipeline 12, the pressure measurer is arranged on the first pipeline 14, and the frequency converter 11 is used for controlling the water pumping quantity of the pump 10; in the invention, different water pumping amounts of the peristaltic pump 10 are controlled by the frequency converter 11, so that the artificial blood vessel 6 generates different pressure values and different liquid flow rate values, and the cell compatibility under different pressures and different flow rates is further detected; the dynamic setting mode of the whole circulatory system can test the cell adhesion condition of the artificial blood vessel 6 under the dynamic condition.
Further, a first joint 7 is arranged at the position where the first pipeline 14 is communicated with the liquid collecting tank 3, a second joint 2 is arranged at the position where the third pipeline 15 is communicated with the liquid collecting tank 3, and the first joint 7 and the second joint 2 are both straight joints or forked multi-way joints; the straight joint can be connected with an artificial blood vessel 6 and carries out the detection process; the multi-way connector can be simultaneously connected with a plurality of artificial blood vessels 6 and can be used for simultaneously detecting, so that the detection efficiency is improved; in addition, the same is true for the bifurcated artificial blood vessel 6.
Furthermore, the first pipeline 14, the second pipeline 13, the third pipeline 15 and the fourth pipeline 12 are all silicone tubes without biotoxicity.
Further, the liquid collecting tank 3 and the culture medium bottle 1 are transparent glass water storage containers; the inside condition of collection liquid tank 3 and culture medium bottle 1 is surveyd in real time to the detection personnel of being convenient for.
Referring to fig. 1, the pressure detector comprises a pressure sensor 9 and a pressure gauge 8, the pressure sensor 9 is arranged on the first pipeline 14 and close to the first joint 7, the pressure sensor 9 is connected with the pressure gauge 8, and the pressure gauge 8 is used for displaying the pressure in the artificial blood vessel 6.
The use method of the artificial blood vessel extracorporeal circulation culture system comprises the following steps:
examples 1,
Referring to FIG. 1, the dynamic cell adhesion status in vitro of the sample of the artificial blood vessel 6, which has a 2mm inner diameter and a 20mm length, is tested under 5kPa, wherein the first connector and the second connector 2 are both one-way connectors.
Closing the water-stop clamp 5, respectively connecting two ends of an artificial blood vessel 6 with the first connector 7 and the second connector 2, injecting complete cell culture solution into the liquid collection tank 3 and immersing the artificial blood vessel 6, after standing culture is carried out for 4-5 hours, opening the water-stop clamp 5, allowing the cell culture solution to enter the culture medium bottle 1, opening the peristaltic pump 10, controlling the flow of the peristaltic pump 10 through the frequency converter 11, and observing the display value of the pressure gauge 8, wherein when the display value is 5kPa, and the variation value of the displayed pressure value within continuous 10 minutes is within the range of 5kPa +/-0.3 kPa of the preset value, the stable preset pressure value is reached; the water stop clamp 5 is opened, and the complete cell culture solution seeped out from the artificial blood vessel 6 enters the culture medium bottle 1 through the fourth pipeline 12; keeping the artificial blood vessel 6 extracorporeal circulation culture system running for a preset time, closing the frequency converter 11 and the peristaltic pump 10, taking out the blood vessel sample, and observing the adhesion and proliferation conditions of cells on the inner wall of the artificial blood vessel 6 sample under the 5kPa dynamic culture condition by utilizing a scanning electron microscope and a fluorescence microscope.
Examples 2,
Referring to FIG. 2, the first connector 7 is a two-way connector and the second connector 2 is a one-way connector, and the dynamic cell adhesion condition in vitro of the bifurcated sample of the artificial blood vessel 6 with an inner diameter of 2mm and a length of 20mm under 5kPa was tested.
Closing the water stop clamp 5, respectively communicating two bifurcated ends of the bifurcated artificial blood vessel 6 with two connectors of a first connector 7, connecting the other end of the bifurcated artificial blood vessel with one connector of a second connector 2, injecting complete cell culture solution into the liquid collection tank 3 and immersing the artificial blood vessel 6, after standing culture for 4 to 5 hours, opening the water stop clamp 5, allowing the cell culture solution to enter a culture bottle, opening the peristaltic pump 10, controlling the flow of the peristaltic pump 10 through the frequency converter 11, observing the display value of the pressure gauge 8, and when the display value is 5kPa, and the change value of the displayed pressure value within continuous 10 minutes is within the range of 5kPa +/-0.3 kPa of a preset value, namely reaching a stable preset pressure value; the water stop clamp 5 is opened, and the complete cell culture solution seeped out from the artificial blood vessel 6 enters the culture medium bottle 1 through the fourth pipeline 12; keeping the artificial blood vessel 6 extracorporeal circulation culture system running for a preset time, closing the frequency converter 11 and the peristaltic pump 10, taking out the blood vessel sample, and observing the adhesion and proliferation conditions of cells on the inner wall of the artificial blood vessel 6 sample under the 5kPa dynamic culture condition by utilizing a scanning electron microscope and a fluorescence microscope.
Examples 3,
Referring to FIG. 3, the in vitro dynamic cell adhesion status of the sample of the artificial blood vessel 6, which has an inner diameter of 2mm and a length of 20mm, and which has the first connector 7 and the second connector 2 both of which are three-way connectors, was tested under the condition of 5 kPa.
Closing the water-stop clamp 5, respectively connecting two ends of three artificial blood vessels with three connectors of a first connector 7 and a second connector 2, injecting complete cell culture solution into the liquid collection box 3 and immersing the artificial blood vessels 6, after standing culture for 4-5 hours, opening the water-stop clamp 5, allowing the cell culture solution to enter a culture bottle, opening the peristaltic pump 10, controlling the flow of the peristaltic pump 10 through the frequency converter 11, observing the display value of the pressure gauge 8, and when the display value is 5kPa, and the variation value of the displayed pressure value within continuous 10 minutes is within the range of 5kPa +/-0.3 kPa as the preset value, reaching a stable preset pressure value; the water stop clamp 5 is opened, and the complete cell culture solution seeped out from the artificial blood vessel 6 enters the culture medium bottle 1 through the fourth pipeline 12; keeping the artificial blood vessel 6 extracorporeal circulation culture system running for a preset time, closing the frequency converter 11 and the peristaltic pump 10, taking out the blood vessel sample, and observing the adhesion and proliferation conditions of cells on the inner wall of the artificial blood vessel 6 sample under the 5kPa dynamic culture condition by using a scanning electron microscope and a fluorescence microscope.
The adaptation according to the actual needs is within the scope of the invention.
It will be evident to those skilled in the art that the invention is not limited to the details of the foregoing illustrative embodiments, and that the present invention may be embodied in other specific forms without departing from the spirit or essential attributes thereof. The present embodiments are therefore to be considered in all respects as illustrative and not restrictive, the scope of the invention being indicated by the appended claims rather than by the foregoing description, and all changes which come within the meaning and range of equivalency of the claims are therefore intended to be embraced therein. Any reference sign in a claim should not be construed as limiting the claim concerned.
Claims (10)
1. The utility model provides an artificial blood vessel extracorporeal circulation culture system, its characterized in that includes converter, artificial blood vessel, pressure measurement ware, communicates in proper order and forms closed loop's culture medium bottle, peristaltic pump and header tank through the pipeline, the pipeline includes first pipeline, second pipeline, third pipeline and fourth pipeline, artificial blood vessel sets up in the header tank and both ends communicate with the one end of first pipeline and third pipeline respectively, the one end of peristaltic pump with first pipeline intercommunication, the other end with the one end intercommunication of second pipeline, the other end of second pipeline with the culture medium intercommunication, the other end of third pipeline with culture medium bottle intercommunication, fourth pipeline one end with culture medium bottle intercommunication, the other end with the header tank intercommunication, install the stagnant water on the fourth pipeline and press from both sides, pressure measurement ware sets up on the first pipeline, the converter is used for controlling the pump water yield of pump.
2. The system of claim 1, wherein a first joint is arranged at a position where the first pipeline is communicated with the liquid collecting tank, and a second joint is arranged at a position where the third pipeline is communicated with the liquid collecting tank.
3. The extracorporeal circulation culture system for artificial blood vessels according to claim 2, wherein the first connector and the second connector are both straight connectors or branched multi-way connectors.
4. The culture system of claim 1, wherein the first, second, third and fourth pipelines are non-biotoxic silicone tubes.
5. The system of claim 1, wherein the liquid collection tank and the culture medium bottle are transparent glass water containers.
6. The system of claim 2, wherein the pressure detector comprises a pressure sensor and a pressure gauge, the pressure sensor is disposed on the first pipeline and near the first joint, the pressure sensor is connected to the pressure gauge, and the pressure gauge is used for displaying the pressure in the artificial blood vessel.
7. The use method of the artificial blood vessel extracorporeal circulation culture system is characterized by comprising the following steps:
step 1: injecting complete cell culture solution into the culture medium bottle, respectively connecting two ends of an artificial blood vessel co-cultured with the cell culture solution with the first joint and the second joint, opening the peristaltic pump, controlling the flow of the peristaltic pump through the frequency converter, and enabling the pressure in the artificial blood vessel and the flow rate of the cell culture solution to reach preset values;
step 2: opening the water stop clamp, and enabling the complete cell culture solution seeped out from the artificial blood vessel to enter the culture medium bottle through a fourth pipeline;
and step 3: keeping the artificial blood vessel extracorporeal circulation culture system running for a preset time, closing the frequency converter and the peristaltic pump, taking out the blood vessel sample, and observing the adhesion and proliferation conditions of cells on the inner wall of the artificial blood vessel.
8. The method for using the artificial blood vessel extracorporeal circulation culture system according to claim 7, wherein the artificial blood vessel culture process in step 1 is as follows: and closing the water stop clamp, respectively connecting the two ends of the artificial blood vessel with the first connector and the second connector, injecting complete cell culture solution into the liquid collecting box, immersing the artificial blood vessel, and performing static culture for 4 to 5 hours.
9. The method for using the culture system of artificial blood vessel in extracorporeal circulation according to claim 7, wherein the apparatus for observing the adhesion and proliferation of cells on the inner wall of the artificial blood vessel in step 3 is scanning electron microscope and fluorescence microscope.
10. The method of claim 7, wherein the pressure value of the artificial blood vessel in step 1 is changed within ± 0.3kPa in a continuous time to reach a stable pressure.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116343568A (en) * | 2023-05-30 | 2023-06-27 | 中国医学科学院阜外医院 | Vascular experimental device and experimental method thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2005337758A (en) * | 2004-05-24 | 2005-12-08 | Tissue Engineering Initiative Co Ltd | Method and instrument for evaluating compliance of artificial blood vessel |
| CN105021510A (en) * | 2015-07-16 | 2015-11-04 | 东华大学 | Controllable-pressure artificial blood vessel overall water permeability tester and using method thereof |
-
2021
- 2021-12-30 CN CN202111653486.5A patent/CN115247126A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2005337758A (en) * | 2004-05-24 | 2005-12-08 | Tissue Engineering Initiative Co Ltd | Method and instrument for evaluating compliance of artificial blood vessel |
| CN105021510A (en) * | 2015-07-16 | 2015-11-04 | 东华大学 | Controllable-pressure artificial blood vessel overall water permeability tester and using method thereof |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116343568A (en) * | 2023-05-30 | 2023-06-27 | 中国医学科学院阜外医院 | Vascular experimental device and experimental method thereof |
| CN116343568B (en) * | 2023-05-30 | 2023-09-05 | 中国医学科学院阜外医院 | Vascular experimental device and experimental method thereof |
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