CN109266533B

CN109266533B - Adipose tissue preparation device and method for preparing adipose tissue using the same

Info

Publication number: CN109266533B
Application number: CN201811153479.7A
Authority: CN
Inventors: 邓贵澧; 徐靖宁; 黄君杰
Original assignee: Zhejiang Quantum Medical Devices Co Ltd
Current assignee: Zhejiang Quantum Medical Devices Co Ltd
Priority date: 2018-09-30
Filing date: 2018-09-30
Publication date: 2022-07-01
Anticipated expiration: 2038-09-30
Also published as: CN109266533A

Abstract

The present invention relates to a container, a device, a kit and a method for preparing adipose tissue from aspirated fatty material, which can be used for injection. In particular, a washing and separating vessel for preparing adipose tissue has a washing chamber for washing aspirated fatty material, the vessel having an inlet and an outlet for passing fatty material into the washing chamber through the inlet and for passing a fluid component of said material out of said chamber through the outlet, comprising within said washing chamber at least one stirring assembly for mechanically forming an emulsion of the fluid component, each stirring assembly comprising an elastic member and at least one ball attached to the elastic member. The method comprises a washing step, an emulsifying step and an emulsion removing step, during which the washing and separating vessel is always in a vertical state with the outlet facing downwards. Compared with the existing process, the preparation process is relatively simple, and can save time, reduce the complexity of the apparatus and reduce the possibility of pollution.

Description

Adipose tissue preparation device and method for preparing adipose tissue using the same

Technical Field

The present invention relates to a device and method for preparing adipose tissue for transplantation from lobular fat extracted by liposuction.

Background

As early as 70 s of the last century, autologous fat transplantation has become a method for confirmation in the orthopedic field, and has been gradually applied to various fields of cosmetic surgery. Autologous fat transplantation requires treatment of the sucked fat, and it is important that red blood cells in the sucked fat and fatty oil components produced from the crushed fat cells must be discharged first. This is because the components after the decomposition of erythrocytes destroy normal tissues, thereby destroying adipocytes and reducing the survival rate of transplantation of fats; while fatty oil is separated from fat cells, it decomposes and forms acidic free fatty acids, which also become a factor for destroying normal tissues and normal fat cells to reduce the survival rate of fat transplantation, and the formation of free fatty acids also causes and increases cell death, which easily increases the probability of secondary infection or becomes a culture base of bacteria, and the fatty oil can remain on the wall and cells of fat cells, like blood membranes, to prevent the supply of oxygen necessary for the survival of transplanted fat cells, resulting in the necrosis of fat cells. For these reasons, it is generally necessary to remove the fatty oil before the fat transplantation, but it is still difficult to completely separate the fat due to the operation conditions and facilities, or a part of the fat is mixed into the transplanted fat, particularly, the fatty oil is accumulated in the lower part of the transplanted space when the transplanted amount is large, which may affect the operation effect, and the symptoms include causing inflammation or damaging tissues, forming a large lump, feeling the lump, etc.

In order to remove red blood cells and fatty oil in the extracted fat, the most original method is to naturally separate the extracted fat by standing in a funnel-shaped container, but the separation of the fatty oil (the fat is separated and then located on the upper layer of the fat) is difficult and time-consuming. Another original treatment method is to pack the extracted fat with cloth and then squeeze the fat with hands to separate impurities, and simultaneously compress the fat by the squeezing force to achieve the purpose of separating and filtering out the fat oil, but the method also takes a lot of time, is complicated in process, has a separation effect related to operators, and more importantly, has the risk of infection after the fat is contacted with hands of people and air.

In order to separate red blood cells and fatty oil safely and efficiently, some technical developments using a syringe-shaped container as a separation device and developments of related devices have been successively reported. The chinese patent application 200480002834.0 discloses a closed loop fat transplantation system, which can be seen from the description of the specification, and the transplantation system mainly aims at a large amount of fat transplantation operations to achieve the purpose that bacterial infection does not occur in the processes of fat extraction, liquid separation and fat transplantation, but the composition relationship among the components of the system is relatively complex, the operation requirement is relatively high, and the popularization and application are not facilitated, and the transplantation system does not consider the problem of fat oil separation. Chinese utility model patent 2006220113167 discloses an autologous fat transplantation adipocyte gathers screening syringe in x, sets up inside and outside piston in the syringe, forms negative pressure lacuna and accomplishes the liposuction through the liposuction machine in intraductal, later need take out this inside and outside piston, insert the former syringe push-and-pull rod that has the flap through special transformation, make the grease spill over through the flap through the centrifugation, then change the syringe push-and-pull rod again and accomplish the injection transplantation of fat. It can be seen that the separation of the fatty oil in the patent technology only depends on centrifugation, the spilled fatty oil is only located at the upper part of the fat and is difficult to separate completely, especially when only a small amount of fat tissue is separated, the accuracy is reduced, and only pure fat tissue can be lost to avoid the mixing of the fatty oil, so that the loss of the fat is caused.

Chinese patent application 201180034851.2 discloses an apparatus and method for preparing adipose tissue, the device structure is shown in figure 1, and comprises at least one washing and separating container 1, wherein the at least one washing and separating container 1 is provided with a washing chamber 101 for washing liposuction materials, the container 1 is provided with an inlet 102 and an outlet 103, for allowing lobular fatty material extracted by fat suction to enter the washing chamber 101 through the inlet 102 and for allowing fluid components in the material to leave the chamber 101 through the outlet 103, and said washing chamber 101 comprises means for forming an emulsion of the fluid components by mechanical agitation, the emulsion forming means being constituted by at least one agitation element 104, such as a ball or the like, for increasing the emulsification of said liquid components when said washing and separating vessel 1 is subjected to mechanical or manual agitation. When the device is used for the preparation of adipocytes, as shown in figure 2, it comprises at least four steps, one being a washing step, the container 1 is kept in vertical position with the outlet 103 facing the ground, repeated washing is carried out by pressure injection of a liquid material, such as sterile saline, into said inner washing chamber 101, to obtain a high-purity cell mass free from any oil, blood and any solution used during the extraction (figure 2 a); secondly, an emulsifying step, in which the container is oriented in a horizontal position, i.e. approximately parallel to the ground, said container 1 being stirred manually or mechanically to promote the emulsification of the fluid components, in particular the oil component with the sterile fluid substance and the blood (fig. 2 b); a third emulsion removal step, arranging the container 1 in a vertical position with respect to the floor with the outlet 103 facing downwards, to obtain stratification of the solid components, including the cell debris, the cells and the solid component of the one or more cell agglomerates floating on the emulsion of fluid components, on the liquid emulsion constituting the fat contained in the washing chamber 101, discharging the emulsion of fluid components (i.e. the oil/liquid) from the washing chamber 101 through the outlet 103 of the washing and separating container 1 (fig. 2 c); fourthly, the solid component removal step, such that the outlet 103 of the vessel 1 is oriented vertically upwards, such that the solid component floats on the wash solution (fig. 2d) to discharge the solid component from the wash and separation vessel.

The above invention patent application promotes the production of emulsion by arranging a plurality of pellets in a container as a stirring device, thereby improving the purity of fat cells, but since the pellets have a certain mass and sink to the bottom when the container is vertically placed, and in this case, the pellets are vibrated and shaken, the stirring action of the pellets is very limited, and thus the container needs to be moved transversely and then stirred, i.e., the above-mentioned emulsification step, which increases the operation difficulty. In practice, the above-mentioned emulsification step is carried out in two ways, firstly by manual operation, which increases the possibility of contamination, and secondly by mechanical operation, in which the device for holding the container is specially designed so that it can provide the function of making the container horizontal and shaking it, since a rotation drive means is required to rotate the container from a vertical position to a horizontal position, which increases the mechanical complexity. Meanwhile, in the whole adipose cell preparation process, at least three times of orientation conversion operations of the container are required, and because the two ends of the container are both connected with the pipelines, the stability of the connecting part of the pipelines and the container is inevitably affected under the condition of multiple orientation conversion operations, so that the pollution possibility is increased. In addition, even if the container is horizontally placed in the emulsification step, the balls can move more intensively at the position close to the ground in the shaking/stirring process due to the gravity of the balls, and the stirring action in the whole container is different, so that the emulsification efficiency needs to be improved.

Therefore, in view of the above technical problems of the prior art, there is still a need for an apparatus and method for preparing adipocytes that can be more conveniently operated and more efficiently emulsified.

Disclosure of Invention

The object of the present invention is to provide a method for preparing adipose tissue for transplantation from aspirated fatty material, such as lobular fat, i.e. fatty material containing large clumps of cells, cells and cell debris, which avoids the above-mentioned drawbacks. The method allows the biological material to be maintained in a completely closed system and the adipose tissue to be prepared with a higher degree of purification through relatively simple operation steps and a relatively high degree of emulsification efficiency.

It is a further object of the present invention to provide adipose tissues, particularly adipocytes, prepared by the above method. It contains less blood and oil, and thus can maintain better surgical effect after implantation.

The above object is achieved by means of a device comprising at least one washing and separating vessel having a washing chamber for washing a sucked fatty material, the vessel having an inlet and an outlet for the fatty material to enter the washing chamber through the inlet and for the fluid component of said material to leave said chamber through the outlet, at least one stirring assembly for mechanically forming an emulsion of the fluid component being comprised in said washing chamber, each stirring assembly comprising an elastic member and at least one ball attached to the elastic member; the method comprises at least the following steps: a washing step of repeatedly washing the fatty material injected into the separation and washing container (1) with a liquid; an emulsifying step of agitating the washing and separating container with the washing chamber injected with the liquid, to promote emulsification of the oil component containing the liquid and the blood in the washing chamber, to form an emulsion; an emulsion removing step of leaving the separation and washing container in which the emulsion is formed standing to form a layer of the emulsion and the solid components floating on the emulsion in the washing chamber, and then discharging the emulsion from the outlet; wherein in the above washing, emulsifying and emulsion removing steps the separation and washing container is always in a vertical position with respect to the ground with the outlet (103) facing downwards.

Optionally, a size reduction device is arranged in the washing chamber, and before, after or during the washing step, a size reduction step of passing the fatty material through the size reduction device at least once is further included.

Preferably, the method further comprises a solid component recovery step after the emulsion is removed. Optionally, in the solid component recovery step, the separation and washing vessel is in a vertical position relative to the ground with the outlet facing upwards, and hydraulic thrust is applied from the inlet to recover the solid component at the outlet.

Optionally, in the emulsifying step, the washing and separating container is agitated in such a manner that the washing and separating container is reciprocally vibrated in a horizontal direction and/or a vertical direction, and/or is rotated centering on a longitudinal axis of the washing and separating container.

Optionally, the process of agitating the washing and separating vessel is effected manually or mechanically.

Optionally, in the emulsion removal step, the emulsion is discharged by means of gravity-induced liquid flow.

Optionally, in the emulsion removal step, the emulsion is forced out by injecting a washing fluid into the washing chamber through the opening.

Preferably, the washing step, the emulsifying step and the emulsion removing step are repeated at least three times.

Optionally, from the emulsion removal step to the solid component recovery step, the washing and separation vessel, via its holding means, effects a switch between a vertical position relative to the ground with the outlet facing downwards to a vertical position relative to the ground with the outlet facing upwards.

The object of the present invention also includes obtaining adipose tissue prepared by the method for preparing adipose tissue as described above.

Compared with the prior art, the invention has the following advantages:

since the elastic member is fixedly connected to the ball, even when the container is in an upright state, slight shaking and shaking causes a large movement of the ball in the container, and the turning of the spring causes an undifferentiated swirling motion, thereby more effectively promoting emulsion formation. Accordingly, the method for preparing adipose tissue according to the present invention maintains the container in a vertical position with the outlet facing downward all the time before forming adipose tissue for transplantation, so that, in the washing, emulsifying, emulsion removing steps and the repeated process of the several steps, there is no need to manually or mechanically adjust the position state of the container, thereby remarkably simplifying the preparation operation process, on one hand improving the work efficiency, on the other hand, since the position of the container is changed by one inversion in the whole preparation process of adipose tissue, the number of manual operations and the pulling torsion between the container and the pipeline in the position changing process are reduced, thereby reducing the contamination risk. When the container is mechanically operated, the method of the invention requires only the operating device to have the function of inverting the container from the vertical position and agitating it, thus also reducing the complexity of the machine in the case of a mechanically driven operation.

Drawings

In order to more clearly illustrate the technical solutions of the present application, the drawings needed for the description of the embodiments or the prior art will be briefly described below, and it is obvious that the drawings in the following description are only some embodiments of the present application, and it is obvious for those skilled in the art to obtain other drawings without creative efforts.

Fig. 1 shows a side view of a washing and separating vessel of a prior art adipose tissue preparation device.

Fig. 2 shows a schematic flow diagram of an operation for preparing adipose tissue based on a preparation device comprising the washing and separating vessel shown in fig. 1.

Fig. 3 shows a schematic longitudinal section of a washing and separating vessel according to a first embodiment of the invention.

Fig. 4 shows a schematic longitudinal section of a washing and separating vessel according to a second embodiment of the invention.

Fig. 5 shows a schematic longitudinal section through a washing and separating vessel according to a third embodiment of the invention.

Fig. 6 shows a schematic longitudinal section of a washing and separating vessel according to a fourth embodiment of the invention.

Fig. 7 shows a schematic longitudinal section of a washing and separating vessel according to a fifth embodiment of the invention.

Fig. 8 shows a schematic view of a fat preparation device kit comprising a washing and separating vessel according to an embodiment of the present invention.

Fig. 9 is a flowchart of a method of preparing adipose tissue using the fat preparation device according to the embodiment of the present invention.

Fig. 10 is a schematic view showing the state of each step of preparing adipose tissues using the fat preparation device according to the embodiment of the present invention. Wherein fig. 10(a), fig. 10(b), fig. 10(c) and fig. 10(d) show schematic diagrams of a washing step, an emulsifying step, an emulsion removing step and a solid component recovering step, respectively.

Detailed Description

The technical solutions in the embodiments of the present application will be clearly and completely described below with reference to the drawings of the specification, and it is obvious that the described embodiments are only a part of the embodiments of the present application, 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 application.

< washing and separating vessel >

< first embodiment >

Fig. 3 shows a schematic longitudinal section of a washing and separating vessel according to a first embodiment of the invention. The washing and separating container 1 has a washing chamber 101 for washing the sucked material, said container 1 having an inlet 102 and an outlet 103 for the sucked fatty material to enter the washing chamber 101 through the inlet 102 and to leave said chamber 101 through the outlet 103 in a time sequence, i.e. in a sequence of first a fluid component and then a solid component, said washing chamber 101 comprising means for mechanically forming an emulsion of the fluid component, in particular of an oil, blood and/or other sterile liquid solution obtained from ruptured fat cells.

The emulsion forming device consists of at least one stirring assembly 4, the at least one stirring assembly 4 comprising an elastic member 41 and a ball 42 attached to the elastic member 41, in particular, the elastic member 41 may be a coil spring. Optionally, the elastic members 41 are secured to the top and bottom of the washing and separating vessel 1. The spheres 42 are attached to the resilient member 41 for increasing the emulsion of the liquid component when the washing and separating vessel 1 is subjected to shaking. In the device of the invention, simple manual stirring of the container 1 can provide separation of the liquid phase, comprising the fluid emulsion, from the solid phase, comprising the cells, the cell fragments, the cell clumps, obtained.

Optionally, the number of stirring assemblies 4 is equal to or greater than 4.

Alternatively, the stirring assemblies 4 are evenly distributed within the vessel 1. For example, symmetrically distributed on the bottom wall and on the top wall of the container 1 with respect to the centre line of the container 1 in the height direction, and/or circumferentially distributed with respect to the longitudinal axis of the container 1.

Optionally, a size reduction device is provided in the washing and separating vessel 1, which size reduction device comprises several size reduction meshes. At least at the inlet 102 of the washing and separating vessel 1 a first size reduction mesh 301 is arranged. The material to be treated passes through the size reduction mesh 301 before entering the washing chamber 101 of the washing and separating container 1 to cut the material into smaller sizes. Optionally, a second size reduction mesh 302 is also provided at the outlet 103 of the washing and separating vessel 1.

Preferably, the mesh size of the size reduction mesh decreases from the inlet 102 to the outlet 103.

The size-reducing net 3 is made up of at least one set of parallel or intersecting sheets or cut lines. Preferably, the sheet or cut line is made of a sterilizing material such as metal.

The mesh openings of the size reduction mesh 34 may have various shapes, such as, but not limited to: triangular, rectangular, diamond, hexagonal, octagonal, etc.

< second embodiment >

A washing and separating vessel in accordance with a second embodiment of the present invention is shown in fig. 4, and has a structure similar to that of the first embodiment except that an elastic member 41 in the form of a coil spring is fixedly attached to a sidewall of the washing and separating vessel 1.

Alternatively, the agitating members are alternately arranged on opposite sides of the inner wall of the washing and separating vessel 1.

Alternatively, the stirring assembly is arranged in a spiral form on the side wall of the washing and separating vessel 1.

In this embodiment, optionally, a third size reduction mesh 303 is provided between the first size reduction mesh 301 and the second size reduction mesh 302, at a position closer to the inlet 102 than to the outlet 103.

The material to be treated passes through the two stage size reduction meshes 301, 303 before entering the washing chamber 101 of the washing and separating vessel 1, whereby the material can be cut to smaller sizes.

< third embodiment >

A washing and separating vessel in accordance with a third embodiment of the invention is shown in fig. 5 and is similar in construction to the first embodiment (the size reduction net is omitted from illustration) except that an elastic member 41 in the form of a coil spring is not connected to the washing and separating vessel 1 but has a free end. In this embodiment, the elastic member 41 provides a sufficient repulsive force with the inner wall of the container 1, so that shaking of the container 1 can cause active movement of the ball 42 regardless of whether the container 1 is in a vertical or horizontal orientation, thereby effectively improving the emulsifying effect.

Optionally, both ends of the elastic member 41 in the form of a coil spring are provided with balls 42.

In this embodiment, optionally, 1 to 3 size-reducing meshes are additionally provided at any position between the first size-reducing mesh 301 and the second size-reducing mesh 302, thereby further improving the size-reducing effect and securing the separation ratio of the adipose tissues.

< fourth embodiment >

A washing and separating vessel in accordance with a fourth embodiment of the present invention is shown in fig. 6, and has a similar structure to that of the first embodiment (size reduction net is omitted from illustration), except that the agitating assembly 4 ' includes an elastic member 41 ' which is a leaf spring, the elastic member 41 ' in the form of a leaf spring is fixedly attached to the inner wall of the vessel, and a ball 42 ' is fixedly attached to the elastic member 41 '.

Alternatively, elastic members 41' in the form of leaf springs are attached to both upper and lower ends of the washing and separating vessel 1, or to the inner wall thereof.

In this embodiment, the orientation of the leaf springs used as the elastic members 41' may be the same, for example, the surfaces of the leaf springs having a larger area face a certain direction, or may be different, for example, the surfaces of the leaf springs having a larger area face the container wall, or may be randomly arranged.

Alternatively, the leaf springs may be of the same length or of different lengths, for example in alternating lengths or in a helically extended form.

< fifth embodiment >

A washing and separating vessel according to a fifth embodiment of the invention is shown in fig. 7 and is similar in construction to the first embodiment (size reduction mesh not shown) except that the agitator assembly 4 "comprises resilient members 41" and spheres 42 ", wherein more than one sphere 42" is attached to each resilient member 41 ".

Alternatively, the elastic member 42 "is a helical spring, which is either attached to the inner wall of the washing and separating container 1 or is free in the washing chamber.

Optionally, more than one ball 42 "is attached to the same side or different sides of the same resilient member 42".

Optionally, the ball 42 "is connected to the resilient member 42" by a short wire.

< adipose tissue preparation device and kit >

Fig. 8 shows a schematic view of a fat preparation device kit comprising a washing and separating vessel according to an embodiment of the present invention. The kit allows the transfer of fatty material to take place in a closed system where the fatty material from the donor and the finally prepared fatty tissue are completely free from the external environment.

The kit comprises a washing and separating container 1 as described in the above embodiments, a syringe 7 for aspirating material from a patient, a syringe 12 for collecting/injecting processed biological material, a washing liquid container 10 (e.g. a bag containing saline), a container 11 for collecting waste material, which are connected or can be connected together by a 7 sterile tube, a multi-way valve, a luer connector.

In particular, the inlet 102 and/or the outlet 103 of the washing and separating container 1 can be connected to a three-way valve 9, such as a luer-lock syringe 7 known in the art connected by a tube or duct 8 to the inlet 102 of the washing and separating container 1 equipped with the three-way valve 9, the syringe 7 performing the aspiration of the fatty material via a cannula provided with one or more holes on its surface.

The washing liquid bag 10 contains a liquid, such as physiological saline, which is used to evacuate air from the container 1, provide pressure to force the fatty material into and out of the chamber 101, and to repeatedly wash the fatty material before washing and separating the fatty material.

The syringe 12 is connected to an outlet 103 for collecting prepared biological material mixed with liquid, which is allowed to settle to obtain separation of solid components from residual liquid components. The material stored in the container 12 may be designed to be reprocessed in washing and separating the container 1 and/or size reducing container 2 prior to final storage and/or re-injection.

A container for collecting waste material, such as a bag 11, is connected to the outlet 103 through the three-way valve 9 to collect the emulsion therein.

The washing and separating vessel 1 may be held by a holding means (not shown) and provide the required driving of the movement of the vessel 1, for example, rotation of the vessel 1 in a plane perpendicular to the ground can be provided to effect inversion from the outlet 103 towards the ground to the inlet 102 towards the ground, and at the same time, vibration driving of the vessel 1 up and down and/or side to side in a plane perpendicular to the ground can also be provided. Alternatively, the holding means may also provide rotation of the container 1 about an axis perpendicular to the ground.

< method for producing adipose tissue >

Fig. 9 and fig. 10(a) -10(d) schematically illustrate a method of preparing adipose tissue using the fat preparation device according to the embodiment of the present invention.

First, liquid is introduced into the container 1, and then lipoaspirate is injected into the washing and separating chamber 101.

In a washing step 501, a liquid material, such as sterile saline, is pressure injected into said washing chamber 101 to repeatedly wash the fat mass injected into the separation and washing container 1, to obtain a high purity cell mass free of any oil, blood and any solution used during the extraction.

In this step, the container is preferably held in an upright position relative to the ground with the outlet 103 facing downwards, as shown in fig. 10 (a).

Optionally, the fatty material is injected into the chamber 101 by means of a suction syringe for extraction, for example a double-syringe equipped with a valve, directly or through a tube connected to the syringe and to the opening of the container, and is pushed into said chamber by the action of the pressure exerted by the piston of the syringe.

Optionally, the fatty material is passed through the size reduction device at least once before, after or simultaneously with the washing step to obtain a cell pellet of a predetermined size.

In an emulsifying step 502, a sanitizing washing solution, such as saline, is injected at least once into the washing chamber 101 containing the fatty materials of the washing and separating vessel 1, which contains at least one stirring assembly 4. The washing and separating container 1 is agitated manually or mechanically, for example, so that the washing and separating container 1 is vibrated in the horizontal direction and/or in the vertical direction, or so that it is rotated, thereby causing a large amplitude movement of the balls 42 of the agitation assembly 4 provided on the elastic member 41, causing an undifferentiated vortex movement of the fluid in the container, so as to promote the emulsification of the oil component containing the sterilizing fluid substance and the blood in the container.

Preferably, in this step, the container is kept in a vertical position with respect to the ground with the outlet 103 facing downwards, as shown in fig. 10 (b).

During this emulsifying step, the openings of the container 1, i.e. the inlet 102 and the outlet 103, are closed to prevent any leakage of material.

In this emulsifying step, the shaking, shaking and rotation of the container 1, due to the presence of the stirring assembly 4, causes a large amplitude movement of the spheres 42 under the action of the elastic force of the elastic member 41, which movement can cause an undifferentiated vortex movement of the fluid inside the container, the washing and separating container 1 containing the fat being thus stirred with a force which does not cause the rupture of the cell walls but is sufficient to form an emulsion, that is, to disperse the tiny oil droplets in the washing fluid, mixing with the sanitizing solution.

In the emulsion removal step 503, the container is still held in a vertical position relative to the ground with the outlet 103 facing downwards, resting to form a stratification of the liquid emulsion of fat and the solid components floating above the emulsion, in particular the solid components comprising cell debris, cells and one or more cell clumps floating on the emulsion of fluid components, within the washing chamber 101, the emulsion components being in contact with the outlet 103 of the washing and separating container 1 in the lower part of the washing chamber 101, as shown in particular in fig. 10 (c). The emulsion of fluid components (i.e. oil and liquid) is discharged from the washing chamber 101 through the outlet 103 of the washing and separating vessel 1.

Alternatively, the discharge of the emulsion is obtained by the flow of physiological fluid induced by gravity.

Alternatively, the emulsion is forced out by injecting a washing fluid through the opening 102 with a given pressure.

The emulsion is collected in a container for collecting waste material, such as bag 11, to prevent contamination of the external environment and the cellular material contained in chamber 101 with the emulsion.

The above washing, emulsifying and emulsion removing steps may be repeated as many times as necessary. The washing and separation is repeated by injecting a sterile washing solution into the chamber, and then forming an emulsion (by agitating the container while closing the inlet and outlet) and draining the emulsion (by injecting a clean washing solution) until the outflowing liquid-oil-blood emulsion appears free of any impurities such as blood and oil.

In the three steps of washing, emulsifying and emulsion removal described above, the washing and separating vessel 1 is preferably always in a vertical position with respect to the ground, with the outlet 103 facing downwards.

After a number of cycles of the three steps or three, the final washing and separation procedure is completed and the solid component material remaining in the chamber 101 of the container 1 is available for implantation.

At the solid component recovering step 504, the solid component available for transplantation is recovered at the outlet 103, preferably by applying a hydraulic thrust from the inlet 102. In this process, the container 1 is preferably brought into a vertical position with respect to the ground, with the outlet 103 facing upwards. Therefore, when the liquid is injected from the inlet 102, the liquid continuously pushes up the solid component floating thereon, and finally the solid component is discharged from the washing and separating vessel 1 via the outlet 103, as shown in fig. 10 (d).

It will be appreciated that when one or more size reduction devices are present in the washing and separating vessel 1, the solid component performs a further size reduction during recovery at least by the size reduction device near the outlet 103.

In the process of carrying out and circulating the steps, the container 1 only needs to be switched between the two states of the outlet 103 facing vertically downwards and the outlet 103 facing vertically upwards, the operation is simple, the processing time can be effectively saved, and the possibility of pollution in the operation process can be reduced.

The solid component obtained by the above-described method of the present invention comprises cell aggregates, single cells and cell debris, which can be used as a bio-filler. Since the whole method does not involve any enzyme or other chemical material, the cells do not undergo any chemical, biological reaction, but only change the size of the cell mass by mechanical action to obtain a cell mass of smaller size and larger exposed surface, which is suitable for transplantation by a micro-fine injection device, so that the obtained cell mass, single cells and cell debris are more suitable for human tissue filling.

The adipose tissues thus prepared can be used for autologous transplantation. It is contemplated that it may also be used for allograft transplantation. For example, adipose tissue prepared using the apparatus and method of the present invention is useful for correction of facial, body volume defects. At the same time, the device and method of the present invention can also be used to provide a larger mass of adipose tissue with stem cells on its surface, the filling of which allows for rapid regeneration of the treated tissue.

The above disclosed devices, kits and methods forming the subject of the present invention can obviously be used not only for the preparation of adipose tissue to be transplanted, but also for the preparation of any type of tissue/cell mass that needs to have a high level of purity for use.

It should be understood by those skilled in the art that the above embodiments are only for illustrating the present invention and are not to be used as a limitation of the present invention, and that changes and modifications to the above embodiments are within the scope of the claims of the present invention as long as they are within the spirit and scope of the present invention.

Claims

1. A method for preparing adipose tissue using a washing and separating vessel (1), said washing and separating vessel (1) having a washing chamber (101) for washing adipose material, an inlet (102) and an outlet (103) for fatty material entering the washing chamber (101) through the inlet (102) and for fluid components of said material exiting the washing chamber (101) through the outlet (103), a plurality of stirring assemblies (4) for an emulsion mechanically forming fluid components being comprised within the washing chamber (101), each stirring assembly (4) comprising an elastic member (41) and at least one ball (42) attached to the elastic member (41), wherein one end of the elastic member (41) is fixedly attached to the ball (42) and the other end of the elastic member (41) is attached to a side wall of the washing and separating vessel (1) while the stirring assemblies are arranged in a spiral form in the washing and separating vessel (1) The side wall of the device (1) or the other end of the elastic component (41) is in a free shape; the method comprises at least the following steps:

a washing step (501) of repeatedly washing the fatty material injected into the washing and separating vessel (1) with a liquid;

an emulsifying step (502) of agitating the washing and separating container (1) with the washing chamber (101) injected with liquid, promoting the emulsification of the fluid components in the washing chamber (101), forming an emulsion;

an emulsion removing step (503) of leaving the washing and separating vessel (1) in which the emulsion is formed standing to form a stratification of the emulsion and the solid components floating on the emulsion in the washing chamber (101), and then discharging the emulsion from the outlet (103);

wherein, in the above washing step (501), emulsifying step (502) and emulsion removing step (503), the washing and separating vessel (1) is always in a vertical position with respect to the ground with the outlet (103) facing downwards.

2. Method for preparing adipose tissue according to claim 1, wherein a size reduction device is provided inside the washing chamber (101), further comprising a size reduction step of passing the adipose material at least once through the size reduction device before, after or during the washing step.

3. The method of preparing adipose tissue according to claim 1 or 2, wherein the method further comprises a solid component recovery step (504) after removing the emulsion.

4. A method for preparing adipose tissue according to claim 3, wherein, in the solid component recovery step (504), the washing and separating vessel (1) is in a vertical position with respect to the ground with the outlet (103) facing upwards, a hydraulic thrust is applied from the inlet (102) to recover the solid component at the outlet (103).

5. The method for preparing adipose tissue according to claim 1 or 2, wherein, in the emulsifying step (502), the washing and separating vessel (1) is agitated in such a manner that the washing and separating vessel (1) is reciprocally vibrated in a horizontal direction or a vertical direction, or is rotated centering on a longitudinal axis of the washing and separating vessel (1).

6. The method for preparing adipose tissue according to claim 5, wherein the process of agitating the washing and separating vessel (1) is achieved manually or mechanically.

7. The method for preparing adipose tissue according to claim 1 or 2, wherein, in the emulsion removing step (503), the emulsion is discharged by means of liquid flow induced by gravity.

8. Method for preparing adipose tissue according to claim 1 or 2, wherein, in the emulsion removal step (503), the emulsion is forced out by injecting a washing fluid into the washing chamber (101) through the inlet (102).

9. The method for preparing adipose tissue according to claim 1 or 2, wherein the washing step (501), the emulsifying step (502), and the emulsion removing step (503) are repeated at least three times.