CN112410179B - Simple in-vitro liver perfusion device for digestion and separation of liver cells - Google Patents
Simple in-vitro liver perfusion device for digestion and separation of liver cells Download PDFInfo
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- CN112410179B CN112410179B CN202011426174.6A CN202011426174A CN112410179B CN 112410179 B CN112410179 B CN 112410179B CN 202011426174 A CN202011426174 A CN 202011426174A CN 112410179 B CN112410179 B CN 112410179B
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M47/00—Means for after-treatment of the produced biomass or of the fermentation or metabolic products, e.g. storage of biomass
- C12M47/04—Cell isolation or sorting
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M23/00—Constructional details, e.g. recesses, hinges
- C12M23/48—Holding appliances; Racks; Supports
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M23/00—Constructional details, e.g. recesses, hinges
- C12M23/52—Mobile; Means for transporting the apparatus
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M29/00—Means for introduction, extraction or recirculation of materials, e.g. pumps
- C12M29/10—Perfusion
Abstract
The invention discloses a simple external liver perfusion device for digesting and separating liver cells, which comprises a liver tissue separation sterile box and a fixing mechanism, wherein the fixing mechanism is arranged in the liver tissue separation sterile box and comprises a left extrusion unit and a right extrusion unit which have the same structure, the liver tissue is placed between the two extrusion units, a puncture needle and a sleeve are inserted into the liver tissue, so that perfusion fluid flows into the liver tissue from the puncture needle, the liver tissue is clamped by the two extrusion units by moving towards each other, a plurality of groups of expansion units are arranged on the sleeve outside the puncture needle, the expansion units are movably connected with a sliding ring through a hinging point by a plurality of pairs of supporting bodies, and when the two extrusion units are mutually close, the expansion units can be automatically opened outwards under the action of thrust, so that the liver tissue is clamped and fixed until the liver tissue is completely separated, the separation time is shortened, and the efficiency of the liver tissue perfusion operation is further improved.
Description
Technical Field
The invention belongs to the technical field related to liver tissue separation of liver cells, and particularly relates to a simple in-vitro liver perfusion device for digesting and separating liver cells.
Background
Hepatocytes are the main functional cells of the liver, consisting mainly of deoxyribonucleic acid and histones. The liver cells are polygonal, have a diameter of about 20-30 per liter (micrometer), have 6-8 faces, and have different sizes under different physiological conditions, such as large volume of liver cells when starved, and can be divided into three faces of sinus face, liver cell face and biliary duct face.
The existing technology of a simple in-vitro liver perfusion device for digesting and separating liver cells has the following problems: when the simple in-vitro liver perfusion device for separating liver cells by digestion is used for separating liver tissues, the liver tissues are needed to be sheared through scissors and then perfusion liquid is introduced, so that the damage to the liver cells is large, the integrity of the cells is poor, meanwhile, the liver cells are not easy to loose, the extraction difficulty is large, and the separation efficiency of the liver cells is influenced.
Disclosure of Invention
The invention aims to provide a simple in-vitro liver perfusion device for digesting and separating liver cells, so as to solve the problems of great damage to the liver cells and great extraction difficulty in the background technology.
In order to achieve the above purpose, the present invention provides the following technical solutions: the simple in-vitro liver perfusion device for digesting and separating liver cells comprises a liver tissue separation sterile box and a fixing mechanism, wherein the fixing mechanism is arranged in the liver tissue separation sterile box and comprises a left extruding unit and a right extruding unit which have the same structure and are symmetrically arranged;
an extrusion space for placing liver tissues is enclosed between the two extrusion units;
a perfusion device is arranged on each extrusion unit and is used for guiding perfusion liquid in the external perfusion bottle into liver tissues in the extrusion space;
the left extrusion unit and the right extrusion unit are connected with elastic pieces, and the elastic pieces are used for enabling the extrusion units on the two sides to fold into the extrusion space and extrude liver tissues.
Preferably, the extrusion units on the left side and the right side of the extrusion space are respectively two mutually parallel sliding plates, and the two sliding plates are relatively closed under the action of the elastic piece.
Preferably, the two groups of liquid filling devices are respectively arranged at the left side and the right side of the liver tissue separation sterile box, each group of liquid filling device comprises a filling bottle, a catheter, a liquid storage box and a puncture needle which are sequentially connected from outside to inside, the puncture needles of the two groups of liquid filling devices are arranged in parallel in a crossing way, and the sliding plate is arranged on the puncture needles in a sliding way.
Preferably, each set of irrigation means comprises at least two needles.
Preferably, the outer wall of the puncture needle is provided with a liquid inlet.
Preferably, the elastic piece comprises a first spring and a fourth spring which are the same, wherein two ends of the first spring are respectively fixed at the upper ends of the extrusion units at two sides, two ends of the fourth spring are respectively fixed at the lower ends of the extrusion units at two sides, and the first spring and the fourth spring are parallel and are in a stretching state.
Preferably, the elastic piece further comprises a third spring fixedly connected between the left liquid storage box and the side wall of the sliding plate and a second spring fixedly connected between the right liquid storage box and the side wall of the sliding plate, and the third spring and the second spring are in a propping state.
Preferably, a sleeve which can expand outwards when the extrusion unit is closed is sleeved outside the puncture needle, one end of the sleeve is fixedly connected with the sliding plate, and the other end of the sleeve is fixedly connected with the end part of the needle head of the puncture needle.
Preferably, the sleeve comprises a plurality of expansion units and a plurality of sliding rings sleeved on the outer wall of the puncture needle in a sliding mode, the adjacent expansion units are connected through one sliding ring, each expansion unit comprises four groups of supporting bodies, and the four groups of supporting bodies are distributed annularly around the axis of the puncture needle.
Preferably, each group of supporting bodies comprises a first supporting sheet and a second supporting sheet which are mutually hinged, the first supporting sheet and the second supporting sheet between adjacent expanding units are hinged at two ends of the same slip ring, the first supporting sheet of the expanding unit positioned at the outermost side is hinged with the end part of the needle head, and the second supporting sheet of the expanding unit positioned at the innermost side is hinged with the sliding plate.
Compared with the existing technology of the simple in-vitro liver perfusion device for digesting and separating liver cells, the invention provides the simple in-vitro liver perfusion device for digesting and separating liver cells, which has the following beneficial effects:
according to the invention, the fixing mechanism consisting of two extrusion units with the same structure is arranged in the liver tissue separation sterile box, the extrusion units are respectively two mutually parallel sliding plates, the two sliding plates are internally provided with the puncture needles which transversely penetrate through the guide pipe and are communicated with the perfusion bottle to form a liver tissue filtrate loop, the perfusion bottle is used for infusing perfusion liquid to the puncture needles, the liver tissues are diluted by the perfusion liquid, so that liver cells are separated conveniently, the two sliding plates are relatively close under the action of the elastic piece, and the puncture needles on the two side extrusion units are parallel and are arranged in a cross way, so that the running paths of the puncture needles on each liquid storage box and the second liquid storage box are not overlapped, the puncture needles are guided by the first sliding plate and the second sliding plate, so that the puncture needles can be inserted into the liver tissues in a staggered way, the liver tissues in the extrusion space are extruded by the first sliding plate and the second sliding plate matched with the two sides, and the separation efficiency of the liver cells in the liver tissues is accelerated;
according to the invention, the sleeve capable of expanding outwards is sleeved outside the puncture needle, one end of the sleeve is fixedly connected with the first sliding plate, the other end of the sleeve is fixedly connected with the end part of the needle head of the puncture needle, a plurality of expansion units and a plurality of sliding rings sleeved on the outer wall of the puncture needle are arranged on the sleeve, the adjacent expansion units are connected through one sliding ring, each expansion unit consists of four groups of supporting bodies distributed annularly, the sliding rings are sleeved on the needle tube body of the puncture needle in a sliding manner, the four groups of supporting bodies are hinged through a plurality of hinge points, and when the sliding rings are blocked by the first sliding plate and the second sliding plate, the four groups of supporting bodies can be expanded, further compress and clamp liver tissues, the separation of the liver tissues is assisted, and the separation efficiency of the liver tissues is improved, so that the separation is more thorough.
Drawings
The accompanying drawings, which are included to provide a further understanding of the invention and are incorporated in and constitute a part of this specification, illustrate and together with the embodiments of the invention and do not constitute a limitation to the invention, and in which:
FIG. 1 is a schematic diagram of a simplified in vitro liver perfusion device for digestion and separation of hepatocytes according to the present invention;
FIG. 2 is a schematic diagram showing the operation of the simple in vitro liver perfusion device for digesting and separating liver cells according to the present invention;
FIG. 3 is an enlarged schematic view of the portion A of FIG. 1 according to the present invention;
FIG. 4 is a schematic view of a deformed cross-section of a sleeve according to the present invention;
in the figure: 1. a conduit; 2. a liquid storage box; 3. a puncture needle; 4. a first slide plate; 5. a sleeve; 501. a first hinge point; 502. a first support sheet; 503. a second hinge point; 504. a second support sheet; 505. a third hinge point; 506. a slip ring; 6. a first spring; 7. a second slide plate; 8. a second spring; 10. a third spring; 12. a spring IV; 13. a liver tissue separation sterile box; 15. and (5) filling the bottle.
Description of the embodiments
The following description of the embodiments of the present invention will be made clearly and completely with reference to the accompanying drawings, in which it is apparent that the embodiments described are only some embodiments of the present invention, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
Examples
In order to clamp liver tissue and separate liver cells from interstitial fluid rapidly, the present invention proposes a first embodiment, referring to fig. 1 and 2, the present invention provides a technical solution: a simple and easy external liver perfusion device for digesting and separating hepatocytes, including hepatic tissue separation aseptic tank 13 and fixed establishment, fixed establishment installs in hepatic tissue separation aseptic tank 13, and fixed establishment is including controlling two extrusion units that the structure is the same, and extrusion unit is two slide that are parallel to each other respectively, is first slide 4 and second slide 7 respectively, after leading in the perfusion liquid to hepatic tissue, two slides are drawn together relatively under the effect of elastic component, extrudes hepatic tissue in the extrusion space, accelerates the separation efficiency of hepatocytes in the hepatic tissue.
A perfusion device is arranged on each extrusion unit for guiding perfusion liquid in the external perfusion bottle 15 into liver tissue in the extrusion space; the perfusion device comprises a catheter 1, a liquid storage box 2, a puncture needle 3 and a perfusion bottle 15, wherein a plurality of liquid inlets are formed in the outer wall of the puncture needle 3, so that perfusion liquid in the perfusion bottle 15 can be discharged into liver tissues sequentially through the catheter 1, the liquid storage box 2 and the liquid inlets formed in the side wall of the puncture needle 3 and the liquid discharge holes in the needle head part, and the liver tissues are diluted by the perfusion liquid, so that liver cells are separated conveniently.
The first sliding plate 4 (or the second sliding plate 7) is provided with a through hole which is convenient for the puncture needle 3 to pass through, one end of the puncture needle 3 passes through the through hole on the first sliding plate 4 (or the second sliding plate 7) and is in through connection with the liquid storage box 2, the outer side outer wall of the liquid storage box 2 is in through connection with one end of the catheter 1, the other end of the catheter 1 is communicated with the perfusion bottle 15, and a liver tissue filtrate loop is formed after the catheter 1 is communicated with the perfusion bottle 15.
Two puncture needles 3 are respectively arranged on the liquid storage boxes 2 at two sides, and the puncture needles 3 at two sides are parallel and are arranged in a cross way, so that the passing routes of the puncture needles 3 on each liquid storage box 2 and the second liquid storage box 9 are not overlapped in a cross way. The first slide plate 4 and the second slide plate 7 are respectively arranged on the puncture needles 3 at two sides in a sliding way, and the puncture needles 3 can also play a role in guiding the first slide plate 4 and the second slide plate 7.
The upper ends of the first slide plate 4 and the second slide plate 7 are respectively and fixedly connected with the two ends of the first spring 6, the lower ends of the first slide plate 4 and the second slide plate 7 are respectively and fixedly connected with the two ends of the fourth spring 12, the first slide plate 4 and the second slide plate 7 tend to slide in a mutually approaching state under the pulling of the elastic force of the first spring 6 and the fourth spring 12, and the first slide plate 4 and the second slide plate 7 are used for clamping liver tissues between the first slide plate 4 and the second slide plate 7, so that liver cells and tissue fluid can be rapidly separated.
In order to further improve the extrusion force of the sliding plates at two sides, a spring III 10 is fixedly connected between the left liquid storage box 2 and the side wall of the first sliding plate 4, a spring II 8 is fixedly connected between the right liquid storage box 2 and the side wall of the first sliding plate 4, and the spring III 10 and the spring II 8 are in a tight pushing state, so that the sliding plates at two sides are extruded.
Through the scheme, the liver tissues in the extrusion space can be irrigated, and then the liver tissues are extruded through the sliding plates at the two sides, so that the separation efficiency of the liver tissues can be improved.
Examples
In order to assist in separating liver tissues and enable the separation to be more thorough, the invention provides another embodiment, a sleeve 5 capable of expanding outwards is sleeved outside the puncture needle 3, one end of the sleeve 5 is fixedly connected with the first sliding plate 4, the other end of the sleeve 5 is fixedly connected with the end part of the needle head of the puncture needle 3, a plurality of expansion units and a plurality of sliding rings 506 sleeved on the outer wall of the puncture needle 3 are arranged on the sleeve 5, adjacent expansion units are connected through one sliding ring 506, each expansion unit comprises four groups of supporting bodies, the four groups of supporting bodies are distributed annularly around the axis of the puncture needle 3, the sliding rings 506 are sleeved on the needle tube of the puncture needle 3 in a sliding mode, and the sliding rings 506 are blocked by the first sliding plate 4 and the second sliding plate 7 to expand the four groups of supporting bodies to further compress and clamp the liver tissues.
It is noted that each group of supporting bodies comprises a first supporting plate 502, a second hinging point 503 and a second supporting plate 504, the first supporting plate 502 and the second supporting plate 504 are movably connected through the second hinging point 503, the second supporting plate 504 and the sliding ring 506 are movably connected through a third hinging point 505, the first supporting plate 502 positioned at the first group of supporting bodies is movably connected with the needle head end of the puncture needle 3 through the first hinging point 501, and the second supporting plate 504 positioned at the tail group of supporting bodies is connected with the first sliding plate 4 or the second sliding plate 7 through the sliding ring 506. The second supporting piece 504 and the sliding ring 506, and the sliding ring 506 and the first sliding plate 4 or the second sliding plate 7 are movably connected through a plurality of hinge points, so that each group of supporting bodies can be opened under the action of the hinge points, thereby assisting the separation of liver tissues.
The working principle and the using flow of the invention are as follows:
the liver tissue is placed between two pressing units, and then the two pressing units are moved toward each other. At this time, the length of the portion of the puncture needle 3 extending out of the first slide plate 4 and the second slide plate 7 is the longest distance, the first support plate 502, the second hinge point 503 and the second support plate 504 on the sleeve 5 are sequentially attached to the puncture needle 3, the puncture needle 3 and the sleeve 5 are inserted into liver tissues, the perfusion fluid in the perfusion bottle 15 flows into the liquid storage box 2 through the catheter 1, and flows into the liver tissues through the puncture needle 3, the distance between the first slide plate 4 and the second slide plate 7 is reduced due to the action of the first spring 6 and the fourth spring 12, the first slide plate 4 and the second slide plate 7 clamp the liver tissues, along with the gradual separation of liver cells in the liver tissues, the liver tissue volume is reduced until the puncture needle 3 on the two extrusion units respectively abuts against the first slide plate 4 or the second slide plate 7 on the opposite sides, the liver tissue volume is further reduced, at this time, the first slide plate 4 applies acting force to the puncture needle 3 on the second slide plate 7 against the second slide plate 7, and simultaneously under the action of the first spring 6 and the fourth spring 12 makes the second slide plate 7 contact with the puncture needle 3 on the outer side of the second slide plate 3, and the puncture needle 3 is expanded, and the acting force is applied to the second slide plate 3 on the second slide plate 3 against the second slide plate 3, and the puncture needle 3 is completely expanded, and the puncture needle 3 is expanded until the acting force is completely on the second slide plate 3 is reached.
Although embodiments of the present invention have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions and alterations can be made therein without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.
Claims (2)
1. A simple and easy external liver perfusion device for digesting and separating hepatocytes, including liver tissue separation aseptic tank (13) and fixed establishment, its characterized in that: the fixing mechanism is arranged in the liver tissue separation sterile box (13) and comprises a left extruding unit and a right extruding unit which have the same structure and are symmetrically arranged; an extrusion space for placing liver tissues is enclosed between the two extrusion units; a perfusion device is arranged on each extrusion unit and is used for guiding perfusion liquid in an external perfusion bottle (15) into liver tissues in the extrusion space; the left extrusion unit and the right extrusion unit are connected with elastic pieces, and the elastic pieces are used for folding the extrusion units at the two sides into the extrusion space to extrude liver tissues; the extrusion units at the left side and the right side of the extrusion space are respectively two mutually parallel sliding plates, and the two sliding plates are relatively closed under the action of the elastic piece; the two groups of liquid filling devices are respectively arranged at the left side and the right side of the liver tissue separation sterile box (13), each group of liquid filling devices comprises a perfusion bottle (15), a catheter (1), a liquid storage box (2) and a puncture needle (3) which are sequentially connected from outside to inside, the puncture needles (3) of the two groups of liquid filling devices are arranged in parallel and in a crossed manner, and a sliding plate is arranged on the puncture needles (3) in a sliding manner; each group of liquid filling devices comprises at least two puncture needles (3); the outer wall of the puncture needle (3) is provided with a liquid inlet; the elastic piece comprises a first spring (6) and a fourth spring (12) which are the same, wherein two ends of the first spring (6) are respectively fixed at the upper ends of the extrusion units at two sides, two ends of the fourth spring (12) are respectively fixed at the lower ends of the extrusion units at two sides, and the first spring (6) and the fourth spring (12) are parallel to each other and are in a stretching state; the elastic piece also comprises a third spring (10) fixedly connected between the left liquid storage box (2) and the side wall of the sliding plate and a second spring (8) fixedly connected between the right liquid storage box (2) and the side wall of the sliding plate, and the third spring (10) and the second spring (8) are in a propping state; a sleeve (5) which can expand outwards when the extrusion units are closed is sleeved outside the puncture needle (3), one end of the sleeve (5) is fixedly connected with the sliding plate, and the other end of the sleeve is fixedly connected with the end part of the needle head of the puncture needle (3); the sleeve (5) comprises a plurality of expansion units and a plurality of sliding rings (506) sleeved on the outer wall of the puncture needle (3) in a sliding mode, the adjacent expansion units are connected through one sliding ring (506), each group of expansion units comprises four groups of supporting bodies, and the four groups of supporting bodies are distributed annularly around the axis of the puncture needle (3).
2. The simple in vitro liver perfusion device for the digestion and separation of hepatocytes of claim 1, wherein: each group of supporting bodies comprises a first supporting sheet (502) and a second supporting sheet (504) which are hinged with each other, the first supporting sheet (502) and the second supporting sheet (504) between adjacent expanding units are hinged at two ends of the same slip ring (506), the first supporting sheet (502) of the expanding unit positioned at the outermost side is hinged with the end part of the needle head, and the second supporting sheet (504) of the expanding unit positioned at the innermost side is hinged with the sliding plate.
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| Application Number | Priority Date | Filing Date | Title |
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| CN202011426174.6A CN112410179B (en) | 2020-12-09 | 2020-12-09 | Simple in-vitro liver perfusion device for digestion and separation of liver cells |
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| Application Number | Priority Date | Filing Date | Title |
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| CN202011426174.6A CN112410179B (en) | 2020-12-09 | 2020-12-09 | Simple in-vitro liver perfusion device for digestion and separation of liver cells |
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| CN112410179B true CN112410179B (en) | 2023-09-29 |
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| CN112410179A (en) | 2021-02-26 |
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