CN112410179A - Simple in-vitro liver perfusion device for digesting and separating liver cells - Google Patents
Simple in-vitro liver perfusion device for digesting and separating liver cells Download PDFInfo
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Abstract
The invention discloses a simple in-vitro 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, the fixing mechanism comprises a left extrusion unit and a right extrusion unit which are identical in structure, liver tissues are placed between the two extrusion units, a puncture needle and a sleeve are inserted into the liver tissues, perfusion liquid flows into the liver tissues from the puncture needle, the liver tissues are clamped through the opposite movement of the two extrusion units, meanwhile, 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 plurality of pairs of supporting bodies through hinge points, when the two extrusion units are close to each other, the expansion units can automatically expand outwards under the thrust action to clamp and fix the liver tissues until the liver tissues are completely separated, the separation time is reduced, thereby further improving the efficiency of the liver tissue perfusion operation.
Description
Technical Field
The invention belongs to the technical field related to hepatic tissue isolation of hepatic cells, and particularly relates to a simple in-vitro hepatic perfusion device for digesting and separating hepatic cells.
Background
Hepatocytes are the main functional cells of the liver, mainly composed of deoxyribonucleic acid and histones. The liver cells are polygonal, have diameter of about 20-30/plus (micrometer) and 6-8 surfaces, and have different sizes under different physiological conditions, such as hepatic cell volume increase during hunger, and each hepatic cell surface can be divided into three types, namely sinusoid surface, hepatic cell surface and bile duct surface.
The existing simple in-vitro liver perfusion device technology for digesting and separating liver cells has the following problems: when the simple in-vitro liver perfusion device for digesting and separating the liver cells is used for separating the liver tissues, the liver tissues are cut by scissors and then introduced into perfusion liquid, 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 loosen, 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 purpose, the invention provides the following technical scheme: the simple in-vitro liver perfusion device for digesting and separating the 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 extrusion unit and a right extrusion unit which have the same structure and are symmetrically arranged;
a pressing space for placing the liver tissue is enclosed between the two pressing units;
each extrusion unit is provided with a perfusion device for guiding perfusion liquid in an external perfusion bottle into the interior of the liver tissue in the extrusion space;
the left extrusion unit and the right extrusion unit are connected with elastic pieces for enabling the extrusion units at the two sides to fold towards the extrusion space and extrude the liver tissue.
Preferably, the extrusion units on the left side and the right side of the extrusion space are two sliding plates which are parallel to each other respectively, 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 devices comprises a flow 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 a parallel and crossed manner, and the sliding plates are arranged on the puncture needles in a sliding manner.
Preferably, each group of the liquid filling devices comprises at least two puncture needles.
Preferably, the outer wall of the puncture needle is provided with a liquid inlet hole.
Preferably, the elastic member includes the same spring one and spring four, and the both ends of spring one are fixed in the upper end of the extrusion unit of both sides respectively, and spring four is fixed in the lower extreme of the extrusion unit of both sides respectively, and spring one and spring four are parallel to each other, and all are tensile state.
Preferably, the elastic part further comprises a third spring fixedly connected between the liquid storage box on the left side and the side wall of the sliding plate, and a second spring fixedly connected between the liquid storage box on the right side and the side wall of the sliding plate, and the third spring and the second spring are in a tightly-pushing 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 manner, 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 annularly distributed around the axis of the puncture needle.
Preferably, each group of support bodies comprises a first support sheet and a second support sheet which are hinged with each other, the first support sheet and the second support sheet between adjacent expansion units are hinged at two ends of the same slip ring, the first support sheet of the expansion unit positioned at the outermost side is hinged with the end part of the needle head, and the second support sheet of the expansion unit positioned at the innermost side is hinged with the sliding plate.
Compared with the prior art of the simple in-vitro liver perfusion device for digesting and separating the liver cells, the invention provides the simple in-vitro liver perfusion device for digesting and separating the liver cells, which has the following beneficial effects:
the invention arranges a fixing mechanism composed of two extrusion units with the same structure in a liver tissue separating sterile box, the extrusion units are two sliding plates which are parallel to each other, puncture needles which transversely penetrate are arranged in the two sliding plates, the puncture needles are communicated with a perfusion bottle through a conduit to form a liver tissue filtering liquid loop, perfusion liquid is infused into the puncture needles through the perfusion bottle, the liver tissue is diluted through the perfusion liquid, the liver cell separation is convenient, the two sliding plates are relatively closed under the action of an elastic part, and the puncture needles on the extrusion units at two sides are parallel and are arranged in a cross way, so that the passing routes of the puncture needles on the liquid storage boxes and the second liquid storage box can not be crossed and overlapped, the puncture needles can be inserted into the liver tissue in a cross way through the first sliding plate and the second sliding plate which are matched, and the liver tissue in the extrusion space can be extruded through the first sliding plate and the second sliding plate which are matched with the two sides, accelerating the separation efficiency of the liver cells in the liver tissue;
the invention can expand to the outside by sleeving the sleeve which can be expanded to the outside on the outside of 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 which are 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 support bodies which are distributed annularly, the sliding rings are sleeved on the needle tube body of the puncture needle in a sliding way, the four groups of support bodies are hinged through a plurality of hinge points, when the sliding rings are blocked by the first sliding plate and the second sliding plate, the four groups of support bodies can be expanded to further compress and clamp the liver tissues, the separation of the liver tissues.
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 embodiments of the invention and together with the description serve to explain the principles of the invention without limiting the invention in which:
FIG. 1 is a schematic structural diagram of a simple extracorporeal liver perfusion apparatus for digesting and separating liver cells according to the present invention;
FIG. 2 is a schematic diagram of the operation process of the simple extracorporeal liver perfusion apparatus for digesting and separating liver cells according to the present invention;
FIG. 3 is an enlarged view of the point A in FIG. 1 according to the present invention;
FIG. 4 is a cross-sectional view of a modified casing according to the present invention;
in the figure: 1. a conduit; 2. a liquid storage box; 3. puncturing 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 fourth spring; 13. a liver tissue separation sterile box; 15. and (4) filling a flow bottle.
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 derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Example 1
In order to clamp the liver tissue and rapidly separate the liver cells from the tissue fluid, the present invention provides a first embodiment, referring to fig. 1 and 2, and the present invention provides a technical solution: a simple and easy external liver perfusion device for digesting separation hepatocyte, including liver tissue separation sterile box 13 and fixed establishment, fixed establishment installs in liver tissue separation sterile box 13, fixed establishment includes the extrusion unit that two structures are the same about, the extrusion unit is two slides that are parallel to each other respectively, be first slide 4 and second slide 7 respectively, after leading-in perfusion liquid in to the liver tissue, two slides draw close relatively under the effect of elastic component, extrude the liver tissue in the extrusion space, the separation efficiency of hepatocyte in the liver tissue accelerates.
A perfusion device is arranged on each extrusion unit and is used for guiding perfusion liquid in the external perfusion bottle 15 into the 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 inlet holes are formed in the outer wall of the puncture needle 3, so that perfusion liquid in the perfusion bottle 15 is discharged to the liver tissue through the liquid inlet holes formed in the side walls of the catheter 1, the liquid storage box 2 and the puncture needle 3 and the liquid outlet hole in the needle head part in sequence, the liver tissue is diluted through the perfusion liquid, and the 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 communicated with the liquid storage box 2, the outer wall of the outer side of the liquid storage box 2 is communicated with one end of the guide pipe 1, and the other end of the guide pipe 1 is communicated with the perfusion bottle 15, so that a liver tissue filtering liquid loop is formed after the guide pipe 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 the liquid storage boxes 2 and the second liquid storage box 9 can not be crossed and overlapped. The first sliding plate 4 and the second sliding plate 7 are respectively arranged on the puncture needles 3 at two sides in a sliding way, and the first sliding plate 4 and the second sliding plate 7 can also be guided by the puncture needles 3.
The upper ends of the first sliding plate 4 and the second sliding plate 7 are fixedly connected with the two ends of the first spring 6 respectively, the lower ends of the first sliding plate 4 and the second sliding plate 7 are fixedly connected with the two ends of the fourth spring 12 respectively, and the first sliding plate 4 and the second sliding plate 7 tend to slide under the state of being close to each other under the pulling of the elastic forces of the first spring 6 and the fourth spring 12 so as to clamp the hepatic tissue between the first sliding plate 4 and the second sliding plate 7 and achieve the purpose of quickly separating the hepatic cells from the tissue fluid.
In order to further improve the extrusion force of the sliding plates on the two sides, a spring III 10 is fixedly connected between the liquid storage box 2 on the left side and the side wall of the first sliding plate 4, a spring II 8 is fixedly connected between the liquid storage box 2 on the right side and the side wall of the first sliding plate 4, and the spring III 10 and the spring II 8 are in a tightly-pushing state so as to extrude the sliding plates on the two sides.
Through above-mentioned scheme, can carry out the perfusion to the liver tissue in the extrusion space, then extrude the liver tissue through the slide of both sides to can improve the separation efficiency of liver tissue.
Example 2
In order to assist the separation of the liver tissue and enable the separation to be more thorough, the invention provides another embodiment, a sleeve 5 which can be expanded to the outside 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 needle head end part of the puncture needle 3, a plurality of expansion units and a plurality of sliding rings 506 which are sleeved on the outer wall of the puncture needle 3 in a sliding mode are arranged on the sleeve 5, adjacent expansion units are connected through one sliding ring 506, each group of expansion units comprises four groups of supporting bodies, the four groups of supporting bodies are distributed in an annular mode around the axis of the puncture needle 3, the sliding rings 506 are sleeved on the needle body 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.
It should be noted that each group of supports includes a first support plate 502, a second hinge point 503 and a second support plate 504, the first support plate 502 and the second support plate 504 are movably connected through the second hinge point 503, the second support plate 504 and the sliding ring 506 are movably connected through a third hinge point 505, the first support plate 502 located in the first group of supports is movably connected with the needle end of the puncture needle 3 through the first hinge point 501, and the second support plate 504 located in the last group of supports is connected with the first sliding plate 4 or the second sliding plate 7 through the sliding ring 506. The second support 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 support bodies can be opened under the action of the hinge points, thereby assisting the separation of liver tissues.
The working principle and the using process of the invention are as follows:
liver tissue is placed between the two compression units, which are then moved towards each other. At this time, the length of the part of the puncture needle 3 extending out of the first sliding plate 4 and the second sliding 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 attached to the puncture needle 3 in sequence, the puncture needle 3 and the sleeve 5 are inserted into the liver tissue, the perfusion liquid in the perfusion bottle 15 flows into the liquid storage box 2 through the conduit 1 and the second conduit 11 and flows into the liver tissue through the puncture needle 3, the distance between the first sliding plate 4 and the second sliding plate 7 is reduced due to the action of the spring I6 and the spring IV 12, the liver tissue is clamped by the first sliding plate 4 and the second sliding plate 7, the volume of the liver tissue is reduced along with the gradual separation of the liver cells in the liver tissue until the puncture needles 3 on the two extrusion units respectively abut against the first sliding plate 4 or the second sliding plate 7 on the opposite side, the volume of the liver tissue is further reduced, and the distance between the first sliding plate 4 and the second sliding, at the moment, the first sliding plate 4 exerts an acting force on the puncture needle 3 on the second sliding plate 7 abutted against the first sliding plate, meanwhile, under the action of the first spring 6 and the fourth spring 12, the puncture needle 3 on the second sliding plate 7 retracts into the second sliding plate 7, so that the support body expands towards the outer side of the puncture needle 3, at the moment, the second sliding plate 7 exerts an acting force on the puncture needle 3 on the first sliding plate 4 abutted against the second sliding plate, meanwhile, under the action of the first spring 6 and the fourth spring 12, the puncture needle 3 on the first sliding plate 4 retracts into the first sliding plate 4, so that the support body expands towards the outer side of the puncture needle 3, all the support bodies begin to expand towards the outer side of the puncture needle 3, and the liver tissue is clamped and fixed until.
Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that changes, modifications, substitutions and alterations can be made in these embodiments without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.
Claims (10)
1. A simple and easy external liver perfusion device for digesting separation hepatocyte, including liver tissue separation aseptic box (13) and fixed establishment, its characterized in that: the fixing mechanism is arranged in a 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;
a pressing space for placing the liver tissue is enclosed between the two pressing units;
each extrusion unit is provided with a perfusion device for guiding perfusion liquid in an external perfusion bottle (15) into the liver tissue in the extrusion space;
the left extrusion unit and the right extrusion unit are connected with elastic pieces for enabling the extrusion units at the two sides to fold towards the extrusion space and extrude the liver tissue.
2. The simplified extracorporeal liver perfusion apparatus for digesting isolated liver cells according to claim 1, wherein: the extrusion units on the left side and the right side of the extrusion space are respectively two sliding plates which are parallel to each other, and the two sliding plates are relatively closed under the action of the elastic piece.
3. The simplified extracorporeal liver perfusion apparatus for digesting isolated liver cells according to claim 2, wherein: the liquid filling devices are two groups and are respectively arranged on the left side and the right side of the liver tissue separation sterile box (13), each group of liquid filling devices respectively comprises a flow filling 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 a parallel and crossed manner, and the sliding plates are arranged on the puncture needles (3) in a sliding manner.
4. The simplified extracorporeal liver perfusion apparatus for digesting isolated liver cells according to claim 3, wherein: each group of the liquid filling devices comprises at least two puncture needles (3).
5. The simplified extracorporeal liver perfusion apparatus for digesting isolated liver cells according to claim 3, wherein: the outer wall of the puncture needle (3) is provided with a liquid inlet hole.
6. The simplified extracorporeal liver perfusion apparatus for digesting isolated liver cells according to claim 1, wherein: the elastic piece comprises a first spring (6) and a fourth spring (12) which are the same, the two ends of the first spring (6) are fixed at the upper ends of the extrusion units on the two sides respectively, the fourth spring (12) is fixed at the lower ends of the extrusion units on the two sides respectively, and the first spring (6) and the fourth spring (12) are parallel to each other and are in a stretching state.
7. The simplified extracorporeal liver perfusion apparatus for digesting isolated liver cells according to claim 6, wherein: the elastic component further comprises a third spring (10) fixedly connected between the liquid storage box (2) on the left side and the side wall of the sliding plate, and a second spring (8) fixedly connected between the liquid storage box (2) on the right side and the side wall of the sliding plate, wherein the third spring (10) and the second spring (8) are in a tightly-pushing state.
8. The simplified extracorporeal liver perfusion apparatus for digesting isolated liver cells according to any one of claims 1 to 7, wherein: the outer portion of the puncture needle (3) is sleeved with a sleeve (5) which can expand outwards when the extrusion unit is closed, 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 portion of the needle head of the puncture needle (3).
9. The simplified extracorporeal liver perfusion apparatus for digesting isolated liver cells according to claim 8, wherein: 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, adjacent expansion units are connected through one sliding ring (506), 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 (3).
10. The simplified extracorporeal liver perfusion apparatus for digesting isolated liver cells of claim 9, 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 expansion units are hinged at two ends of the same sliding ring (506), the first supporting sheet (502) of the expansion unit positioned at the outermost side is hinged with the end part of the needle head, and the second supporting sheet (504) of the expansion unit positioned at the innermost side is hinged with the sliding plate.
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