CN112755370A - Mesenchymal stem cell exosome endometrium preparation, preparation method and uterine cavity perfusion tube - Google Patents
Mesenchymal stem cell exosome endometrium preparation, preparation method and uterine cavity perfusion tube Download PDFInfo
- Publication number
- CN112755370A CN112755370A CN202110012672.4A CN202110012672A CN112755370A CN 112755370 A CN112755370 A CN 112755370A CN 202110012672 A CN202110012672 A CN 202110012672A CN 112755370 A CN112755370 A CN 112755370A
- Authority
- CN
- China
- Prior art keywords
- mesenchymal stem
- preparation
- stem cell
- tube
- exosome
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 210000002901 mesenchymal stem cell Anatomy 0.000 title claims abstract description 70
- 210000001808 exosome Anatomy 0.000 title claims abstract description 62
- 238000002360 preparation method Methods 0.000 title claims abstract description 60
- 210000004696 endometrium Anatomy 0.000 title claims abstract description 32
- 230000010412 perfusion Effects 0.000 title claims abstract description 26
- 239000006166 lysate Substances 0.000 claims abstract description 17
- 230000002357 endometrial effect Effects 0.000 claims abstract description 13
- 239000003634 thrombocyte concentrate Substances 0.000 claims abstract description 8
- 210000004027 cell Anatomy 0.000 claims abstract description 5
- 239000008280 blood Substances 0.000 claims abstract description 4
- 210000004369 blood Anatomy 0.000 claims abstract description 4
- 238000005119 centrifugation Methods 0.000 claims abstract description 4
- 230000008014 freezing Effects 0.000 claims abstract description 4
- 238000007710 freezing Methods 0.000 claims abstract description 4
- 238000002156 mixing Methods 0.000 claims abstract description 4
- 238000010257 thawing Methods 0.000 claims abstract description 4
- 239000003814 drug Substances 0.000 claims description 21
- 239000007788 liquid Substances 0.000 claims description 10
- 238000004113 cell culture Methods 0.000 claims description 6
- 239000007921 spray Substances 0.000 claims description 6
- 210000000130 stem cell Anatomy 0.000 claims description 6
- 229940079593 drug Drugs 0.000 claims description 3
- 238000000703 high-speed centrifugation Methods 0.000 claims description 3
- 239000004017 serum-free culture medium Substances 0.000 claims description 3
- 238000005199 ultracentrifugation Methods 0.000 claims description 3
- 230000000694 effects Effects 0.000 abstract description 12
- 230000006378 damage Effects 0.000 abstract description 7
- 208000027418 Wounds and injury Diseases 0.000 abstract description 4
- 208000014674 injury Diseases 0.000 abstract description 4
- 239000012528 membrane Substances 0.000 abstract description 2
- 230000002265 prevention Effects 0.000 abstract description 2
- 230000000717 retained effect Effects 0.000 abstract description 2
- 241000700159 Rattus Species 0.000 description 24
- 239000000499 gel Substances 0.000 description 13
- 230000009286 beneficial effect Effects 0.000 description 5
- 230000035935 pregnancy Effects 0.000 description 5
- 238000005452 bending Methods 0.000 description 3
- 230000014759 maintenance of location Effects 0.000 description 3
- 210000001161 mammalian embryo Anatomy 0.000 description 3
- 230000035755 proliferation Effects 0.000 description 3
- 210000004291 uterus Anatomy 0.000 description 3
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 230000004069 differentiation Effects 0.000 description 2
- 235000013601 eggs Nutrition 0.000 description 2
- 210000002257 embryonic structure Anatomy 0.000 description 2
- 229940011871 estrogen Drugs 0.000 description 2
- 239000000262 estrogen Substances 0.000 description 2
- 238000011156 evaluation Methods 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 238000000034 method Methods 0.000 description 2
- 210000001215 vagina Anatomy 0.000 description 2
- 208000005189 Embolism Diseases 0.000 description 1
- 206010061218 Inflammation Diseases 0.000 description 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 1
- 210000001015 abdomen Anatomy 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 230000010267 cellular communication Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 230000032692 embryo implantation Effects 0.000 description 1
- 230000002124 endocrine Effects 0.000 description 1
- 201000006828 endometrial hyperplasia Diseases 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000002349 favourable effect Effects 0.000 description 1
- 239000003102 growth factor Substances 0.000 description 1
- 238000007490 hematoxylin and eosin (H&E) staining Methods 0.000 description 1
- 230000005847 immunogenicity Effects 0.000 description 1
- 230000007365 immunoregulation Effects 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 230000004054 inflammatory process Effects 0.000 description 1
- 230000005906 menstruation Effects 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 238000010992 reflux Methods 0.000 description 1
- 238000007790 scraping Methods 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- 238000005507 spraying Methods 0.000 description 1
- 230000001360 synchronised effect Effects 0.000 description 1
- 230000017423 tissue regeneration Effects 0.000 description 1
- 238000002054 transplantation Methods 0.000 description 1
- 210000000626 ureter Anatomy 0.000 description 1
Images
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
- A61M31/00—Devices for introducing or retaining media, e.g. remedies, in cavities of the body
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61D—VETERINARY INSTRUMENTS, IMPLEMENTS, TOOLS, OR METHODS
- A61D7/00—Devices or methods for introducing solid, liquid, or gaseous remedies or other materials into or onto the bodies of animals
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- 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
- C12N5/0602—Vertebrate cells
- C12N5/0652—Cells of skeletal and connective tissues; Mesenchyme
- C12N5/0662—Stem cells
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
- A61M2210/00—Anatomical parts of the body
- A61M2210/14—Female reproductive, genital organs
- A61M2210/1433—Uterus
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2509/00—Methods for the dissociation of cells, e.g. specific use of enzymes
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Zoology (AREA)
- General Health & Medical Sciences (AREA)
- Wood Science & Technology (AREA)
- Veterinary Medicine (AREA)
- Biotechnology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Animal Behavior & Ethology (AREA)
- Organic Chemistry (AREA)
- Chemical & Material Sciences (AREA)
- Public Health (AREA)
- Genetics & Genomics (AREA)
- Developmental Biology & Embryology (AREA)
- Cell Biology (AREA)
- Hematology (AREA)
- Rheumatology (AREA)
- Microbiology (AREA)
- Anesthesiology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- Heart & Thoracic Surgery (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Prostheses (AREA)
Abstract
An endometrium preparation of mesenchymal stem cell exosome, a preparation method and a uterine cavity perfusion tube are used for prolonging the acting time of exosome in a uterine cavity and achieving the aim of comprehensively treating endometrial injury. The inner membrane preparation comprises a mesenchymal stem cell exosome and a buffer gel, wherein the ratio of the mesenchymal stem cell exosome to the buffer gel is 1-5: 1. the preparation method comprises the following steps of obtaining platelet lysate: collecting whole blood of a patient, obtaining a platelet concentrate through gradient centrifugation, repeatedly freezing and thawing the platelet concentrate, and then centrifuging to remove cell debris to obtain a platelet lysate; obtaining a mesenchymal stem cell exosome; preparing an endometrium preparation of the exosome of the mesenchymal stem cells: and fully mixing the mesenchymal stem cell exosomes with the buffer gel to obtain the mesenchymal stem cell exosome endometrium preparation, and storing at 4 ℃. The invention has good backflow prevention effect, and the preparation can be retained on the inner wall of the uterine cavity for a long time, thereby ensuring the repair effect.
Description
Technical Field
The invention relates to the technical field of medical instruments, in particular to an endometrial preparation containing mesenchymal stem cells exosomes, a preparation method and a uterine cavity perfusion tube.
Background
Successful embryo planting does not leave good quality embryos and good endometrium. In the last 30 years, with the continuous development of the assisted reproduction technology, an embryo culture system is optimized, the embryo selection evaluation is enhanced, and the planting rate is further improved. However, recent data studies have shown that endometrial damage remains a major bottleneck limiting the increase in pregnancy rates despite having good quality embryos for transplantation.
The uterine cavity perfusion can promote the proliferation of endometrium and is more beneficial to nidation of fertilized eggs. The uterine cavity is generally infused after the menstruation is clear, which accords with the physiological cycle of women. The uterine cavity perfusion is mainly used for women with endometrial hyperplasia, and aims to promote endometrial proliferation, so that the endometrium is more favorable for nidation of fertilized eggs, and the conception probability is improved. At present, estrogen is mostly adopted for uterine cavity perfusion to adjust the thickness of an inner membrane in clinic, however, liquid medicine rapidly flows back and out of the uterine cavity, so that the retention time of the medicine in the uterine cavity is too short, the whole uterine cavity cannot be covered, and the expected treatment purpose is difficult to achieve. In addition, estrogen has certain interference effect on endocrine and certain side effect. Mesenchymal stem cells are immature primitive cells with self-renewal and multidirectional differentiation potential, the effects of the mesenchymal stem cells on tissue repair are commonly known in the medical field, and platelet lysate has different degrees of beneficial effects on the proliferation and induced differentiation of the stem cells. Stem cells are in cellular communication primarily through exosomes, which are smaller in size and have no immune rejection concerns compared to mesenchymal stem cells. The stem cell exosome perfused into the uterine cavity can repair and protect damaged endometrium, inhibit inflammation, has the function of immunoregulation, can improve the receptivity of endometrium, and is beneficial to embryo implantation.
Disclosure of Invention
The invention aims to provide an endometrial preparation of mesenchymal stem cell exosomes, a preparation method and a uterine cavity perfusion tube, which are used for prolonging the action time of exosomes in a uterine cavity and achieving the aim of comprehensively treating endometrial injury.
The technical scheme adopted by the invention for solving the technical problems is as follows: the mesenchymal stem cell exosome endometrium preparation is characterized by comprising a mesenchymal stem cell exosome and a buffer gel, wherein the ratio of the mesenchymal stem cell exosome to the buffer gel is 1-5: 1.
a preparation method of a mesenchymal stem cell exosome endometrial preparation is characterized by comprising the following steps:
(1) platelet lysate acquisition: collecting whole blood of a patient, obtaining a platelet concentrate through gradient centrifugation, repeatedly freezing and thawing the platelet concentrate, and then centrifuging to remove cell debris to obtain a platelet lysate;
(2) obtaining the exosome of the mesenchymal stem cell: adding 10% of platelet lysate into a serum-free culture medium for subculturing mesenchymal stem cells to obtain a P2-P6 generation mesenchymal stem cell culture, collecting the mesenchymal stem cell culture, centrifuging to remove stem cells, and performing high-speed centrifugation or ultracentrifugation to obtain a mesenchymal stem cell exosome stimulated by the platelet lysate;
(3) preparing an endometrium preparation of the exosome of the mesenchymal stem cells: and fully mixing the mesenchymal stem cell exosomes with the buffer gel to obtain the mesenchymal stem cell exosome endometrium preparation, and storing at 4 ℃.
A mesenchymal stem cell exosome intrauterine preparation uterine cavity perfusion tube is characterized by comprising:
the outer pipe is a hollow round pipe, a handle is fixed on the outer wall of the outer pipe, and a pair of limiting grooves are formed in the inner wall of the outer pipe;
the inner tube is arranged on the inner side of the outer tube and comprises a flow guide tube, a pressurizing tube and a dosing tube which are sequentially arranged, the flow guide tube is a hose, an elastic sheet is arranged on the side wall of the flow guide tube, the flow guide tube keeps an arc-shaped state under the action of the elastic sheet, the first end of the flow guide tube is a guide part, and the guide part is provided with a spray hole; the pressurizing pipe comprises a corrugated pipe fixedly connected with the second end of the flow guide pipe and a push rod in threaded connection with the first end of the dosing pipe, the second end of the corrugated pipe is fixedly connected with the first end of the push rod, the first end of the corrugated pipe is provided with a mounting ring, and the outer wall of the mounting ring is provided with a limiting block matched with the limiting groove; a one-way valve is arranged in the push rod, liquid can only enter the corrugated pipe from the push rod under the action of the one-way valve, and a filling mechanism for pushing the push rod to move to one side of the corrugated pipe is arranged between the push rod and the outer pipe; the second end of the dosing pipe is an expansion part, and an end cover is arranged at the expansion part.
Furthermore, a connecting ring is arranged at the second end of the flow guide pipe and is in threaded connection with the flow guide pipe, and a mounting ring is arranged at the first end of the corrugated pipe and is in threaded connection with the connecting ring.
Further, the guide portion is hemispherical.
Further, the filling mechanism comprises a first ear plate fixed on the side wall of the outer pipe, a trigger hinged with the first ear plate, a driving gear installed at the first end of the trigger in a one-way rotating mode, a spring seat fixed on the handle, a sliding plate arranged in a groove of the sliding plate on the handle in a sliding mode, a first spring arranged between the sliding plate and the spring seat, and a connecting rod fixed on the sliding plate.
Furthermore, the side wall of the push rod is provided with a long groove, and teeth in contact with the driving gear are arranged in the long groove.
Furthermore, the medicine adding device also comprises an opening mechanism, wherein the opening mechanism comprises a driving rod fixedly connected with the valve core of the one-way valve, a driving fork fixed at the tail end of the driving rod, a driving ring fixed on the inner wall of the medicine adding pipe and a pair of driving blocks fixed on the end surface of the driving ring, and when the medicine adding pipe is rotated, the driving blocks rotate and then are contacted with the driving fork, so that the driving rod is extruded to move to one side of the one-way valve, and the valve core moves in the one-way valve to realize the conduction of the one-.
The invention has the beneficial effects that: the mesenchymal stem cell exosome endometrium preparation, the preparation method and the uterine cavity perfusion tube provided by the invention have good backflow prevention effect, and the preparation can be retained on the inner wall of the uterine cavity for a long time, so that the repair effect is ensured. When in use, the end cover is detached at first, and then the medicine feeding pipe is rotated to conduct the one-way valve. Then the preparation is added into the medicine adding pipe through the flaring part, and then the preparation enters the corrugated pipe through the medicine adding pipe and the push rod in sequence. When the bellows is filled with the reagent (the bellows is the largest in length at the moment), the dosing tube is rotated reversely to close the check valve, and the end cover is arranged on the flared part. The operator holds the handle, stretches into the patient vagina with the outer tube to continue to promote to the palace intracavity, get into the palace intracavity until the guide part, promote the inner tube to the palace intracavity afterwards until the stopper reaches the spacing groove second end (initial state stopper is located the first end of spacing groove, and the first end of spacing groove is the one end that is close to the bellows), make most of honeycomb duct expose from the outer tube. At this time, the flow guide pipe is bent under the action of the elastic sheet, at this time, the driving gear is meshed with the teeth at the first end of the long groove, as shown in fig. 11, the trigger can be pulled at this time to push the push rod to move to the side where the corrugated pipe is located (at this time, the mounting ring and the first end of the corrugated pipe are stationary under the action of the limiting block), and then the corrugated pipe is extruded to enable the preparation in the corrugated pipe to be ejected through the spray holes in the guide part, so that the preparation is poured. Because the honeycomb duct is in a bending state, the whole inner wall of the uterine cavity can be filled without dead angles.
Drawings
FIG. 1 is a graph showing the reflux of the drug solution in rats in the control group and the experimental group;
FIG. 2 is a front view of the uterine cavity perfusion tube;
FIG. 3 is a cross-sectional view of FIG. 2;
FIG. 4 is a cross-sectional view of the outer tube;
FIG. 5 is a cross-sectional view of the inner tube;
FIG. 6 is a cross-sectional view of the draft tube;
FIG. 7 is a cross-sectional view of the pressurization pipe;
FIG. 8 is a cross-sectional view of the dosing tube;
FIG. 9 is an axial view of the drive ring;
FIG. 10 is a cross-sectional view of the trigger;
FIG. 11 is a state diagram of the present invention in use;
in the figure: the device comprises an outer tube 1, a limit groove 10, a handle 11, a spring seat 12, a first spring 121, a first lug plate 13, a trigger 14, a second lug plate 15, a chute 151, a connecting rod 16, a protrusion 161, a sliding plate 162, a through hole 17, a driving gear 18, a locking block 19, a guide pipe 2, a guide part 21, a spring plate 22, a connecting ring 23, a corrugated tube 24, a mounting ring 241, a limit block 242, a push rod 25, an elongated slot 251, a medicine adding tube 26, an expanding part 261, an end cover 262, a check valve 3, a valve core 31, a second spring 32, a driving rod 33, a driving fork 34, a driving ring 4, a driving block 41 and a round hole 42.
Detailed Description
The mesenchymal stem cell exosome endometrial preparation, the preparation method and the uterine cavity perfusion tube of the invention are described in detail below.
The mesenchymal stem cell exosome endometrium preparation comprises a mesenchymal stem cell exosome and a buffer gel, wherein the ratio of the mesenchymal stem cell exosome to the buffer gel is 1-5: 1, namely when the weight part of the buffer gel is 1, the weight part of the mesenchymal stem cell exosome is 1-5. The following describes a method for preparing an endometrial preparation containing mesenchymal stem cells exosomes.
(3) Platelet lysate acquisition: collecting whole blood of a patient, obtaining a platelet concentrate through gradient centrifugation, repeatedly freezing and thawing the platelet concentrate, and then centrifuging to remove cell debris to obtain a platelet lysate; the components such as platelet lysate growth factor and the like are high in content and have no immunogenicity;
(4) obtaining the exosome of the mesenchymal stem cell: adding 10% of platelet lysate into a serum-free culture medium for subculturing mesenchymal stem cells to obtain a P2-P6 generation mesenchymal stem cell culture, collecting the mesenchymal stem cell culture, centrifuging to remove stem cells, and performing high-speed centrifugation or ultracentrifugation to obtain a mesenchymal stem cell exosome stimulated by the platelet lysate;
(3) preparing an endometrium preparation of the exosome of the mesenchymal stem cells: fully mixing the mesenchymal stem cell exosomes with the buffer gel to obtain a mesenchymal stem cell exosome endometrium preparation, and storing at 4 ℃; the ratio of the mesenchymal stem cell exosomes to the buffer gel is as follows: the mesenchymal stem cell exosome endometrium preparation is in a liquid state at the temperature of 1-5: 1 and 4 ℃, and can effectively keep the activity of exosome. Wherein the buffer gel is temperature sensitive gel, colorless transparent liquid at low temperature (0-20 ℃), and stable gel state at high temperature (20-40 ℃). Buffer gels are prior art and will not be described further herein.
Preparing an endometrium preparation of a rat mesenchymal stem cell exosome by taking a rat as a sample, and performing the following experiment on the treatment effect of the endometrium preparation of the mesenchymal stem cell exosome by taking the rat as an experimental object:
(1) establishing a rat endometrium damage model: selecting female SD rats of 8 weeks old, scraping and damaging the left and right uteruses of the rats simultaneously by using a rat endometrial brush, and randomly dividing the rats into two groups, namely a control group and an experimental group, wherein each group comprises 10 rats;
(2) two groups of rats were treated differently: injecting 1.5ml of mesenchymal stem cell exosome preparation into the uterine cavity of a rat in an experimental group; injecting normal saline with equal dosage into uterine cavity of rat in control group; the rats in the experimental group and the rats in the control group are continuously infused in the uterine cavity for 5 days;
(3) analyzing the backflow condition after perfusion: collecting the liquid medicine flowing out of the uterine cavity by using a miniature collector for 30min, measuring the volume, and calculating the average return flow of rats in each group; the statistical data are shown in figure 1, the outflow rate of the liquid medicine in the uterine cavity of the rat in the experimental group is relatively less, which shows that the mesenchymal stem cell exosome preparation can greatly prolong the retention time of the liquid medicine in the uterine cavity;
(4) on day 6, 4 rats (8 uteruses) are randomly selected from a control group and an experimental group respectively, uterus samples are collected and subjected to HE staining, and the thickness of the endometrium is measured under a 200-time microscope to evaluate the damage recovery condition; the evaluation results are shown in table 1;
table 1: endometrium thickness of rats in experimental group and control group
(5) The remaining 6 rats of the control group and the experimental group were mated with normal male rats on day 6, female rats were sacrificed 15 days after the occurrence of vaginal embolus, whether intrauterine pregnancy was established or not was confirmed by opening the abdomen, and the pregnancy rates of the two groups of rats were compared, and the results are shown in table 2.
Table 2: pregnancy of rats in experimental and control groups
The conclusion is that the method integrates the following experiments: the preparation taking the mesenchymal stem cell exosome stimulated by the platelet lysate as the main active ingredient can prolong the retention time of the medicament in the uterine cavity, has excellent repairing effect on the endometrial injury, and is beneficial to improving the pregnancy rate.
When the uterine cavity perfusion treatment is carried out on a human body, the liquid mesenchymal stem cell exosome endometrium preparation is filled in a uterine cavity perfusion tube, and the uterine cavity perfusion is carried out according to a clinical uterine cavity perfusion operation guide. After the endometrium of the mesenchymal stem cell exosome is filled in a uterine cavity, the endometrium preparation of the mesenchymal stem cell exosome is converted into a stable gel state under the in-vivo environment of about 37 ℃, and can be tightly attached to the endometrium for injury repair.
In order to promote the mesenchymal stem cell exosome endometrium preparation to be uniformly sprayed on the inner wall of the uterine cavity, the invention designs the uterine cavity perfusion tube. As shown in fig. 2 to 11, the uterine cavity perfusion tube includes an outer tube 1 and an inner tube. As shown in fig. 2 to 4, the outer tube 1 is a circular plastic tube, a handle 11 and a first ear plate 13 are fixed to a sidewall of the outer tube, a trigger 14 is hinged on the first ear plate, a driving gear 18 is arranged at the first end of the trigger in a unidirectional rotation way, the side wall of the outer tube is provided with a through hole 17, a part of the driving gear passes through the through hole and then extends into the inner side of the outer tube, a connecting rod 16 is arranged between the trigger and the handle, a spring seat 12 is fixed on the handle, a sliding plate 162 is fixed at the first end of the connecting rod, a first spring 121 is arranged between the sliding plate and the inner wall of the spring seat, a sliding plate groove is arranged on the handle, the sliding plate is arranged in the sliding plate groove in a sliding way, a protrusion 161 is fixed to the second end of the link, a second lug 15 is fixed to the trigger, a sliding groove 151 is formed in the second lug, the protrusion is slidably disposed in the sliding groove, and the distance between the trigger and the handle is the largest under the action of the first spring. The handle is held to the user during the operation, pulls out the trigger and can overcome the elastic force effect of first spring and make the trigger around the pin joint swing, and drive gear rotates along with the trigger is synchronous this moment. The spring holder is columniform hollow structure, and one end is uncovered, and the open end and the handle threaded connection of spring holder.
The inner tube is disposed inside the outer tube and is axially movable relative to the outer tube. As shown in fig. 2, 3 and 5, the inner tube mainly comprises a flow guide tube 2, a pressurizing tube and a dosing tube 26, and as shown in fig. 5 and 6, the flow guide tube 2 is a circular tube. The first end of the draft tube is a guide part 21 with a hemispherical structure, an elastic sheet 22 is arranged in the side wall of the draft tube, and the elastic sheet 22 is in a bending state under a natural state. The honeycomb duct is a hose, and after the elastic pieces are inserted into the side wall of the honeycomb duct, the honeycomb duct is in a bent state under the action of the upper and lower groups of elastic pieces. And a connecting ring 23 is arranged at the second end of the draft tube, and the connecting ring 23 is in threaded connection with the draft tube. The arrangement of the connecting ring also limits the elastic sheet in the side wall of the flow guide pipe. The guide part is provided with a plurality of spray holes so as to facilitate the spraying of the preparation.
As shown in fig. 5 and 7, the pressurizing pipe includes a bellows 24 and a push rod 25, and a mounting ring 241 is provided at a first end of the bellows for being threadedly coupled to the connection ring. The second end of the corrugated pipe is fixedly connected with the first end of the push rod. The push rod 25 is a cylindrical hollow structure, and a long groove 251 is formed in the side wall of the push rod, and teeth are arranged in the long groove and used for being in contact with the driving gear. As shown in fig. 3, when the driving gear rotates with the trigger, the teeth on the driving gear contact with the teeth in the elongated slot, and further the push rod is pushed to move to the side where the corrugated pipe is located, and further the corrugated pipe is squeezed to reduce the volume in the corrugated pipe, so that the liquid pressure in the corrugated pipe is increased, and the preparation in the corrugated pipe is forced to enter the flow guide pipe and is ejected through the spray holes on the guide part. The cylindrical outer wall of the mounting ring is provided with a pair of limiting blocks 242, the inner wall of the outer pipe is provided with a pair of limiting grooves 10 matched with the limiting blocks, and the corrugated pipe can be limited by the arrangement and the matching of the limiting grooves and the limiting blocks. The trigger, the driving gear, the connecting rod, the first spring and the spring seat form a filling mechanism.
As shown in fig. 7, a check valve 3 is arranged in the push rod, a valve core 31 is arranged in the check valve 3, and the valve core 31 is contacted with the inner wall of a conical hole in the check valve, so that the check valve is blocked. A second spring 32 is arranged between the valve core and the one-way valve, and the valve core is contacted with a conical hole in the one-way valve under the action of the second spring. In order to realize the one-way passage of the one-way valve, a driving rod 33 is fixed on the valve core, a U-shaped driving fork 34 is fixed at the tail end of the driving rod, and when the driving fork moves, the valve core can be driven by the driving rod to move towards one side where the large end of the inner conical hole of the one-way valve is located, so that the one-way valve is communicated. For this purpose, as shown in fig. 8, a drive ring 4 is fixed to the inner wall of the dosing tube, as shown in fig. 9, a plurality of circular holes 42 are uniformly provided in the circumferential direction on the drive ring, and a pair of drive blocks 41 are provided on the drive ring, the drive blocks being provided with slopes. As shown in fig. 5, the drive fork is naturally in contact with the end surface of the drive ring. When the medicine ureter is added in the rotation, can be so that the drive ring is rotatory, and then make the inclined plane contact on drive fork and the driver block, and then extrude the drive fork through the driver block. As shown in fig. 8, the dosing tube 26 is a circular tube, and a first end of the dosing tube is screwed with a second end of the push rod, and a sealing ring is arranged between the first end and the second end. The second end of the dosing tube is provided with a flared part 261, the flared part is provided with an end cover 262, and the flared part is sealed through the end cover. When in use, the preparation is added into the medicine adding pipe through the flaring part. After pulling the trigger, drive gear rotates around the pin joint, and under the effect of first spring, for avoiding the tooth in the tooth's socket to interfere with the resetting of drive gear when the trigger resets, as shown in fig. 10, be equipped with locking piece 19 between trigger and drive gear, the point portion of locking piece stretches into between the profile of tooth of drive gear. A torsion spring is arranged between the locking block and the trigger, and the driving gear can only rotate in a single direction under the action of the locking block. I.e. the drive gear can only rotate clockwise relative to the trigger and not counter-clockwise, similar to a ratchet and ratchet mechanism in mechanical principles. The driving rod, the driving fork, the driving ring and the driving block form an opening mechanism for opening the one-way valve.
When in use, the end cover is detached at first, and then the medicine feeding pipe is rotated to conduct the one-way valve. Then the preparation is added into the medicine adding pipe through the flaring part, and then the preparation enters the corrugated pipe through the medicine adding pipe and the push rod in sequence. When the bellows is filled with the reagent (the bellows is the largest in length at the moment), the dosing tube is rotated reversely to close the check valve, and the end cover is arranged on the flared part. The operator holds the handle, stretches into the patient vagina with the outer tube to continue to promote to the palace intracavity, get into the palace intracavity until the guide part, promote the inner tube to the palace intracavity afterwards until the stopper reaches the spacing groove second end (initial state stopper is located the first end of spacing groove, and the first end of spacing groove is the one end that is close to the bellows), make most of honeycomb duct expose from the outer tube. At this time, the flow guide pipe is bent under the action of the elastic sheet, at this time, the driving gear is meshed with the teeth at the first end of the long groove, as shown in fig. 11, the trigger can be pulled at this time to push the push rod to move to the side where the corrugated pipe is located (at this time, the mounting ring and the first end of the corrugated pipe are stationary under the action of the limiting block), and then the corrugated pipe is extruded to enable the preparation in the corrugated pipe to be ejected through the spray holes in the guide part, so that the preparation is poured. Because the honeycomb duct is in a bending state, the whole inner wall of the uterine cavity can be filled without dead angles.
Claims (8)
1. The mesenchymal stem cell exosome endometrium preparation is characterized by comprising a mesenchymal stem cell exosome and a buffer gel, wherein the ratio of the mesenchymal stem cell exosome to the buffer gel is 1-5: 1.
2. a preparation method of a mesenchymal stem cell exosome endometrial preparation is characterized by comprising the following steps:
(1) platelet lysate acquisition: collecting whole blood of a patient, obtaining a platelet concentrate through gradient centrifugation, repeatedly freezing and thawing the platelet concentrate, and then centrifuging to remove cell debris to obtain a platelet lysate;
(2) obtaining the exosome of the mesenchymal stem cell: adding 10% of platelet lysate into a serum-free culture medium for subculturing mesenchymal stem cells to obtain a P2-P6 generation mesenchymal stem cell culture, collecting the mesenchymal stem cell culture, centrifuging to remove stem cells, and performing high-speed centrifugation or ultracentrifugation to obtain a mesenchymal stem cell exosome stimulated by the platelet lysate;
(3) preparing an endometrium preparation of the exosome of the mesenchymal stem cells: and fully mixing the mesenchymal stem cell exosomes with the buffer gel to obtain the mesenchymal stem cell exosome endometrium preparation, and storing at 4 ℃.
3. A mesenchymal stem cell exosome intrauterine preparation uterine cavity perfusion tube is characterized by comprising:
the outer pipe is a hollow round pipe, a handle is fixed on the outer wall of the outer pipe, and a pair of limiting grooves are formed in the inner wall of the outer pipe;
the inner tube is arranged on the inner side of the outer tube and comprises a flow guide tube, a pressurizing tube and a dosing tube which are sequentially arranged, the flow guide tube is a hose, an elastic sheet is arranged on the side wall of the flow guide tube, the flow guide tube keeps an arc-shaped state under the action of the elastic sheet, the first end of the flow guide tube is a guide part, and the guide part is provided with a spray hole; the pressurizing pipe comprises a corrugated pipe fixedly connected with the second end of the flow guide pipe and a push rod in threaded connection with the first end of the dosing pipe, the second end of the corrugated pipe is fixedly connected with the first end of the push rod, the first end of the corrugated pipe is provided with a mounting ring, and the outer wall of the mounting ring is provided with a limiting block matched with the limiting groove; a one-way valve is arranged in the push rod, liquid can only enter the corrugated pipe from the push rod under the action of the one-way valve, and a filling mechanism for pushing the push rod to move to one side of the corrugated pipe is arranged between the push rod and the outer pipe; the second end of the dosing pipe is an expansion part, and an end cover is arranged at the expansion part.
4. The perfusion tube for the uterine cavity of claim 3, wherein the second end of the flow guide tube is provided with a connection ring, the connection ring is in threaded connection with the flow guide tube, the first end of the corrugated tube is provided with a mounting ring, and the mounting ring is in threaded connection with the connection ring.
5. The uterine cavity perfusion tube of a mesenchymal stem cell exosome preparation according to claim 3, wherein the guide part is hemispherical.
6. The perfusion tube for the uterine cavity of an exosome preparation of mesenchymal stem cells according to claim 1, which is characterized in that the perfusion mechanism comprises a first ear plate fixed on the side wall of the outer tube, a trigger hinged with the first ear plate, a driving gear unidirectionally and rotatably installed at the first end of the trigger, a spring seat fixed on the handle, a sliding plate slidably arranged in a sliding plate groove on the handle, a first spring arranged between the sliding plate and the spring seat, and a connecting rod fixed on the sliding plate, wherein the side wall of the trigger is provided with a second ear plate, the end part of the connecting rod is provided with a protrusion, the protrusion is slidably arranged in a sliding groove on the second ear plate, and a part of the driving gear passes through a through hole on the side wall of the outer tube and then contacts.
7. The uterine cavity perfusion tube of mesenchymal stem cell exosome and endometrium preparation according to claim 3, wherein the side wall of the push rod is provided with a long groove, and the long groove is internally provided with teeth which are contacted with the driving gear.
8. The perfusion tube for the uterine cavity of the endometrial preparation of the mesenchymal stem cells according to claim 3, which is characterized by further comprising an opening mechanism, wherein the opening mechanism comprises a driving rod fixedly connected with a valve core of the one-way valve, a driving fork fixed at the tail end of the driving rod, a driving ring fixed on the inner wall of the drug adding tube and a pair of driving blocks fixed on the end surface of the driving ring, when the drug adding tube is rotated, the driving blocks rotate to be in contact with the driving fork, and then the driving rod is extruded to move towards one side of the one-way valve, so that the valve core moves in the one-way valve to realize the conduction of the one-way valve.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110012672.4A CN112755370B (en) | 2021-01-06 | 2021-01-06 | Mesenchymal stem cell exosome endometrial preparation, preparation method and uterine cavity perfusion tube |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110012672.4A CN112755370B (en) | 2021-01-06 | 2021-01-06 | Mesenchymal stem cell exosome endometrial preparation, preparation method and uterine cavity perfusion tube |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN112755370A true CN112755370A (en) | 2021-05-07 |
| CN112755370B CN112755370B (en) | 2022-10-21 |
Family
ID=75699926
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202110012672.4A Active CN112755370B (en) | 2021-01-06 | 2021-01-06 | Mesenchymal stem cell exosome endometrial preparation, preparation method and uterine cavity perfusion tube |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN112755370B (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112760293A (en) * | 2021-01-08 | 2021-05-07 | 广东工业大学 | Method for preparing high-activity exosomes by 3D culture of MSC stem cells by using xeno-free serum |
| CN112870228A (en) * | 2021-01-20 | 2021-06-01 | 杭州贤石生物科技有限公司 | Multifunctional microenvironment protection exosome hydrogel and preparation method and application thereof |
Citations (13)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20050137575A1 (en) * | 2003-12-18 | 2005-06-23 | Duke University | Minimally invasive injection devices and methods |
| CN202104958U (en) * | 2011-06-23 | 2012-01-11 | 李洪湘 | Medical suction injection and endoscopic variceal ligation therapeutic apparatus |
| CN202740611U (en) * | 2012-06-21 | 2013-02-20 | 邵光英 | Gun-shaped cervix applicator |
| CN203988487U (en) * | 2014-06-13 | 2014-12-10 | 大庆市兴牧科技有限公司 | Veterinary's uterine irrigation pipe |
| CN205924677U (en) * | 2016-06-06 | 2017-02-08 | 邢磊 | Annotate feeder and annotate appetite controlling means thereof |
| CN206072400U (en) * | 2016-08-16 | 2017-04-05 | 圣博莱阀门有限公司 | A kind of solid forging ramp type ball valve |
| CN207500544U (en) * | 2017-10-09 | 2018-06-15 | 陕西航天动力高科技股份有限公司 | A kind of spherical shape plugging type check valve |
| KR101917748B1 (en) * | 2018-05-25 | 2018-11-12 | 전상표 | Trigger type instrument for artificial insemination |
| CN208785565U (en) * | 2018-03-21 | 2019-04-26 | 林高阳 | A kind of Bladder perfusion of medicine pipe |
| CN110585118A (en) * | 2019-09-30 | 2019-12-20 | 广东华夏健康生命科学有限公司 | Temperature-sensitive gel preparation of endometrial stem cells and preparation method and application thereof |
| CN111021989A (en) * | 2019-12-10 | 2020-04-17 | 中国石油集团川庆钻探工程有限公司长庆井下技术作业公司 | Underground controllable one-way valve and control method |
| CN211434741U (en) * | 2020-01-08 | 2020-09-08 | 重庆市中医院 | Ointment smearing device |
| CN112076416A (en) * | 2020-09-10 | 2020-12-15 | 丁良超 | Insulated temperature-sensing self-starting fire extinguisher |
-
2021
- 2021-01-06 CN CN202110012672.4A patent/CN112755370B/en active Active
Patent Citations (13)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20050137575A1 (en) * | 2003-12-18 | 2005-06-23 | Duke University | Minimally invasive injection devices and methods |
| CN202104958U (en) * | 2011-06-23 | 2012-01-11 | 李洪湘 | Medical suction injection and endoscopic variceal ligation therapeutic apparatus |
| CN202740611U (en) * | 2012-06-21 | 2013-02-20 | 邵光英 | Gun-shaped cervix applicator |
| CN203988487U (en) * | 2014-06-13 | 2014-12-10 | 大庆市兴牧科技有限公司 | Veterinary's uterine irrigation pipe |
| CN205924677U (en) * | 2016-06-06 | 2017-02-08 | 邢磊 | Annotate feeder and annotate appetite controlling means thereof |
| CN206072400U (en) * | 2016-08-16 | 2017-04-05 | 圣博莱阀门有限公司 | A kind of solid forging ramp type ball valve |
| CN207500544U (en) * | 2017-10-09 | 2018-06-15 | 陕西航天动力高科技股份有限公司 | A kind of spherical shape plugging type check valve |
| CN208785565U (en) * | 2018-03-21 | 2019-04-26 | 林高阳 | A kind of Bladder perfusion of medicine pipe |
| KR101917748B1 (en) * | 2018-05-25 | 2018-11-12 | 전상표 | Trigger type instrument for artificial insemination |
| CN110585118A (en) * | 2019-09-30 | 2019-12-20 | 广东华夏健康生命科学有限公司 | Temperature-sensitive gel preparation of endometrial stem cells and preparation method and application thereof |
| CN111021989A (en) * | 2019-12-10 | 2020-04-17 | 中国石油集团川庆钻探工程有限公司长庆井下技术作业公司 | Underground controllable one-way valve and control method |
| CN211434741U (en) * | 2020-01-08 | 2020-09-08 | 重庆市中医院 | Ointment smearing device |
| CN112076416A (en) * | 2020-09-10 | 2020-12-15 | 丁良超 | Insulated temperature-sensing self-starting fire extinguisher |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112760293A (en) * | 2021-01-08 | 2021-05-07 | 广东工业大学 | Method for preparing high-activity exosomes by 3D culture of MSC stem cells by using xeno-free serum |
| CN112760293B (en) * | 2021-01-08 | 2023-04-18 | 广东工业大学 | Method for preparing high-activity exosomes by 3D culture of MSC stem cells by using xeno-free serum |
| CN112870228A (en) * | 2021-01-20 | 2021-06-01 | 杭州贤石生物科技有限公司 | Multifunctional microenvironment protection exosome hydrogel and preparation method and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN112755370B (en) | 2022-10-21 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN112755370B (en) | Mesenchymal stem cell exosome endometrial preparation, preparation method and uterine cavity perfusion tube | |
| CN208404652U (en) | A kind of gynemetrics, which checks, uses device for sampling secretions | |
| Júnior et al. | Anatomical and histological characterization of the cervix in Santa Inês hair ewes | |
| CN111493939A (en) | Protective sampling swab for large animals | |
| CN205612515U (en) | Be used for hemostatic device in uterus | |
| CN203139357U (en) | Gynecological triple-bag multi-cavity tube | |
| CN209826915U (en) | Artificial insemination device for assisted reproduction technology | |
| CN203971144U (en) | Salpingography recanalization conduit | |
| CN208838220U (en) | A kind of milk cow ovarian cyst puncture needle | |
| CN201752400U (en) | Uterine cavity examination resection hysteroscopy | |
| CN106691696B (en) | A tampon for extracting stem cells from menstrual blood | |
| CN213372326U (en) | Endometrium cleaning brush | |
| CN210673896U (en) | Gynaecology washes stick and gynaecology washing unit | |
| CN205697854U (en) | A kind of cytopathology inspection sampling apparatus | |
| CN112890932A (en) | Artificial insemination transplanting device for reproductive medicine | |
| CN211862858U (en) | Cervical mucus swab | |
| CN219022939U (en) | Portable medicine plug administration device | |
| CN107213453B (en) | Abortion medicine | |
| CN215384186U (en) | Novel vagina and cervix cell lavage and recovery integrated device | |
| CN211460453U (en) | Self-service insemination appliance | |
| CN201524345U (en) | Medicine feeder | |
| CN214284995U (en) | Novel gynecological curettage therapeutic device | |
| CN217244722U (en) | Embryo transfer assembly | |
| CN219022227U (en) | Eye nursing washing and disinfecting appliance | |
| CN212491124U (en) | Clinical device of dosing of gynaecology and obstetrics |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20240307 Address after: 250100 workshop 102, building 12, SME industrialization base, biomedical park, 1777 Dazheng Road, high tech Zone, Jinan City, Shandong Province Patentee after: TAIPINGYANG KANGTAI SCIENTIFIC APPARATUS (JINAN) CO.,LTD. Country or region after: China Address before: 250000 805-1, sunshine big sister building, Huaiyin District, Jinan City, Shandong Province Patentee before: Shandong Meiyi Biotechnology Co.,Ltd. Country or region before: China |
|
| TR01 | Transfer of patent right |