EP3743507A1

EP3743507A1 - Device and method for dissociating tissue

Info

Publication number: EP3743507A1
Application number: EP18704159.5A
Authority: EP
Inventors: Christian Reis; Stefan Scheuermann; Armin Schäfer; Daniel EDINGER
Original assignee: Fraunhofer Gesellschaft zur Forderung der Angewandten Forschung eV
Current assignee: Fraunhofer Gesellschaft zur Forderung der Angewandten Forschung eV
Priority date: 2018-01-24
Filing date: 2018-01-24
Publication date: 2020-12-02
Also published as: CN111902528A; US20210140856A1; WO2019145029A1

Abstract

The invention relates to a device and a method for dissociating tissue. In one embodiment, the device comprises at least one dissociation unit for receiving a tissue sample, which is arranged at least partially in a pot-shaped cell sieve. The invention also relates in an embodiment to a device and a method for dissociating tissue, wherein the tissue is dissociated by means of a grinding mechanism having rows of teeth which can be moved relative to one another and which, when moving past one another in the one direction, cut the tissue and, when moving past one another in the opposite direction, grind the tissue.

Description

Apparatus and method for the dissociation of tissue

The invention relates to an apparatus and a method for the dissociation of tissue. In one embodiment, the device has at least one dissociation mechanism for receiving a tissue sample, which is arranged at least partially in a pot-shaped cell sieve. The invention also relates, in one embodiment, to an apparatus and method for tissue disassembly wherein the tissue is dissociated by a grinder with mutually movable rows of teeth which, when moving in one direction, cut the tissue and move it in the opposite direction to rub the cloth.

The dissociation of human or animal tissue poses a challenge for several reasons. Not only are tissues of different origins differing in their consistency, but one and the same tissue sample can also be very heterogeneous. Therefore, it is not yet succeeded in developing a method with which any kind of tissue can be dissociated.

There are several commercially available systems that can disrupt tissues and cells. A reliable method for tissue dissociation into a single cell suspension exists only conditionally.

Object of the present invention is to provide an apparatus and a method for the dissociation of tissue, with which any type of tissue can be dissociated and preferably to a single cell suspension ver works.

The object is achieved by the tissue dissociation devices of claim 1 and claim 6 and by the tissue dissociation method of claim 24. The respective dependent claims set forth advantageous developments of the tissue dissociation devices and the tissue dissociation process ,

According to the invention, an apparatus for the dissociation of tissue is provided which has at least one dissociation unit for receiving a tissue sample and at least one pot-shaped cell sieve. At least one dissociation unit is at least partially arranged in the pot-shaped cell strainer.

A dissociation plant is understood to mean a device to which the tissue can be dissociated. In particular, the dissociation may include cell disruption and / or tissue dissociation. In particular, the process of dissociation of tissue may advantageously comprise attrition of tissue and / or production of single-cell suspension and / or isolation of molecules.

According to the invention, the device for dissociating tissue in this embodiment has a cup-shaped cell strainer. It should be understood by a cup shape in the most general sense, a concave shape into which the dissociation is at least partially introduced. In particular, under a pot shape advantageously a cylindrical shape with one side open End face or front side are understood. The term "cylindrical shape" may be understood in the mathematical most general sense and may include cylinders with any base surface, wherein the cylindrical shape preferably has a straight cylinder axis, but advantageously the cylindrical shape has a circular base surface, so that under a cylindrical shape in the context of the invention Advantageously, a circular cylindrical shape can be understood with unilaterally open end face or end face.

If the cell sieve is cylindrical in this sense, advantageously the dissociation plant also has a circular cross-section. It can then be arranged at least partially in the cell strainer with a coaxial to a cylindrical axis of the kreiszylin derförmigen cell strainer with a circular cross-section in its center axis.

Preferably, the dissociation plant is protected in the cell sieve against rotation relative to the cell sieve.

The cell strainer advantageously has at least one surface which acts as a sieve for the tissue constituents produced by the dissociation plant. In this case, a mesh size of the screen depending on the tissue by the dissociation he testified tissue components may be suitably chosen. Particularly advantageous is a cell strainer, which has a circular cylindrical cup shape with an open and a closed end face or end face, wherein the closed end face or end face and the lateral surface are formed by respective sieves. The sieves can be advantageously carried by a frame. The frame can specify the outer shape of the cell siebes. If the cell sieve has a cylindrical shape, then the frame advantageously runs along edges of the cylindrical shape, whereby the sections of the frame extending along the edges of the cylindrical shape can be connected to one another by struts, which can preferably run parallel to the cylinder axis. This shape of the frame is advantageous for both circular cylindrical screens, as well as cylindrical screens with other bases. The frame can be advantageously made of plastic.

In an advantageous embodiment of the invention, the device for Dissociation of tissue at least one vessel for receiving a tissue sample have. The vessel may be part of the dissociation plant.

Advantageously, a grinder is arranged in the vessel, with which the tissue sample can be dissociated. In this case, the feature may advantageously comprise at least two mutually movable rows of teeth, each having a plurality of teeth. The grinder can thus have at least a first row of teeth with a plurality of teeth and at least one second row of teeth with a plurality of teeth. Vorteilhafterwei se are then adjacent to the rows of teeth at such a distance zueinan arranged that cause the teeth of each adjacent rows of teeth in a distance range around their minimum distance from each other a dissociation of a portion of the tissue sample. Advantageously, therefore, the at least one first row of teeth can be movable relative to the at least one second row of teeth, so that the teeth of the respective first row of teeth move past the teeth of the respective second row of teeth. One of the at least one first rows of teeth and one of the at least one second rows of teeth can then be arranged adjacent to each other at such a distance that the teeth of these adjacent rows of teeth, when moved past each other, into a distance range about their minimum distance from one another Dissociation of a part of the tissue sample cause.

Advantageously, the vessel may have a bottom and a lid. The bottom and the cover can be designed so that they together form the vessel or put together. But it is also possible that the bottom and lid cover opposite sides of a cylindrical shell, so that soil, cover and shell together form the vessel. In an advantageous embodiment, the vessel may be cylindrical, particularly preferably circular cylindrical. Particularly preferred then the rows of teeth are arranged concentrically around a cylinder axis of the vessel.

In an advantageous embodiment of the invention, the teeth of the tooth-rich can be designed so that when they move in a direction past each other, cut the part of the tissue sample and if they move past each other in a direction opposite to this direction, rubbing the part of the tissue sample.

The term "grinder" may include both cutting and trimming devices, so the term "crusher" may be used as an alternative to the term "grinder".

Advantageously, the grinder may have at least two mutually BE wegbare rows of teeth. Each of the rows of teeth then has a more number of teeth. The term "row of teeth" should in this case generally describe a set of teeth that move together in the comminution process.The teeth of a row of teeth may advantageously be arranged along a line which may have different shapes be arranged a circular closed line.

Advantageously, adjacent rows of teeth are arranged at such a distance from one another that the teeth of these adjacent rows of teeth cause a dissociation of a portion of the tissue sample in a distance range about their minimum distance from each other. The distance between two rows of teeth can be defined here either as the distance of the lines along which the teeth of a row of teeth are arranged or as the distance of the teeth at the point of their respective closest approach. Have the teeth of two adjacent rows of teeth different distances in their respective closest approach, so can be defined as the distance and an average distance of the teeth measured at the point of closest approach of each two teeth defi ned.

The distance between the rows of teeth is advantageously dimensioned so that the teeth cause a dissociation of a part of the tissue sample when they are moved past each other. The respectively dissociated part of the tissue sample is that part on which the two teeth, which move past each other, act in a crushing manner. The dissociation takes place in a Abstandsbe the teeth from each other by the minimum distance of the teeth vonei nander instead. Thus, when moving past each other, two teeth of the adjacent rows of teeth move towards each other, starting from a At a distance, an action on the tissue to dissociate the tissue that persists until the teeth have passed a certain maximum distance from each other after they have passed.

It should be noted that the teeth of the adjacent rows of teeth do not have to be assigned to one another in pairs. Advantageously, it is rather the case that the teeth of the adjacent rows of teeth pass one another after each other.

Advantageously, the teeth are configured to cut the tissue as they move in a direction past one another and grind the tissue as they move past each other in opposite directions. To achieve this effect, the teeth can have a variety of different shapes. Advantageously, the teeth may have a cutting edge on one side in the direction of movement and have on their opposite side a part of a milling or tearing device in their direction of motion, so that two teeth of adjacent rows of teeth meeting with this side will cause the tissue to break down ken.

In two adjacent rows of teeth, which together cause a dissociation of the tissue, while the cutting edges and the frictional edges of the two rows of teeth are advantageous opposing orien benefits. If the two rows of teeth move past each other in the opposite direction, they cause the cutting with the two cutting edges in one direction and the rubbing with the fretting sides in the opposite direction. In general, one can define a cutting direction and a rubbing direction that are respectively opposite for cooperating adjacent rows of teeth, i. the cutting directions of the adjacent rows of teeth are opposite to each other and the Zerreibrichtungen the adjacent rows of teeth are entgegenge sets.

For other applications, the teeth may also have other cross-sections ha ben, such as cylindrical or have a karoförmige, parallelogram or diamond-shaped base. In an advantageous embodiment, the rows of teeth are circular and con centric to each other configured. That is, the teeth of a row of teeth are each lined up along a circle and these circles are con centric to each other. The cutting direction described and the trimming described are then advantageously tangential or parallel to the ent speaking circular lines.

In an advantageous embodiment of the invention, the device for the dissociation of tissue may further comprise a reagent vessel, in the opening of the dissociation plant and / or the vessel for receiving the Gewe can be arranged sample. In this case, the reagent vessel z. B. be a glass jar or a plastic container or be made of other suitable Materi alien. Advantageously, the reagent vessel may have a cylindrical portion, the length of which in the direction of its cylinder axis is longer than a diameter of the reagent vessel. The cylindrical shape can have on one side the opening in which the dissociation unit and / or the vessel can be arranged.

In an advantageous embodiment of the invention arranged in the lid of the vessel rows of teeth relative to the reagent vessel can be movable and arranged in the bottom of the vessel rows of numbers can be ge compared to the reagent vessel fixed. In the normal case, the vessel will be used in the Publ tion of the reagent vessel and there, for example, be screwed. Therefore, in this embodiment, the rows of teeth fixed relative to the reagent vessel should be fixed in the state in which the vessel is arranged for intended use in the reagent vessel. The mobility of the teeth arranged in the cover can then be effected by suitable design of the vessel.

In an advantageous embodiment of the invention, the components of the grinder, so the rows of teeth, be designed as interchangeable inserts. These inserts can be inserted into the vessel. The rows of teeth Kings nen be arranged on two plates, so that the teeth are arranged between the plates in a common plane. The idea according to the invention is also realized by a device which has at least one vessel for receiving a tissue sample, and a grinder arranged in the vessel, wherein the grinder has at least two mutually movable rows of teeth. For the vessel, the grinder and the rows of teeth advantageously the above can also be true true.

In this embodiment of the invention, each of the rows of teeth may have a plurality of teeth, wherein adjacent rows of teeth are arranged in a sol chen distance from each other, that the teeth of each of these adjacent rows of teeth in a distance range to their minimum from each other was a dissociation of a part effecting the tissue sample, wherein the teeth are configured to intersect the portion of the tissue sample as they move past each other in one direction and, as they move past each other in that direction, to grind the portion of the tissue sample. It should be noted that in this embodiment of the invention, the cup-shaped cell strainer is not essential. The ge called Dissoziationswerk may advantageously be the grinder in this Ausgestal device.

Also in this embodiment, the device can aufwei sen a reagent vessel, in whose opening said vessel and the grinder can be arranged. It can also be here arrange the arrange in the lid of the vessel ten rows of teeth relative to the reagent vessel and be arranged in the Bo arranged in the vessel rows of teeth relative to the reagent vessel. What has been said above about the reagent vessel can apply here accordingly.

The following statements may be advantageous for all embodiments of the invention.

In an advantageous embodiment, if appropriate, the cell strainer and / or the dissociation unit and / or the vessel and / or the grinding mechanism can be arranged at least partially within a closure cap of the reagent vessel. Advantageously, then the cap can embrace the Publ tion of the reagent vessel. In this case, the cell sieve, the dissociation mechanism, the vessel and / or the grinding mechanism can be as described above be arranged the opening of the reagent vessel. When the closure cap engages around the opening of the reagent vessel, the corresponding elements can at the same time be arranged at least partially within the closure cap. The fact that the closure cap encompasses the opening of the reagent vessel may mean, in particular, that the closure cap encloses or comprises the opening. Particularly advantageously, the cell strainer, the Dissozia tionswerk, the vessel and / or the grinder be fastened to the cap or be clamped or clamped by means of the cap between an edge of the reagent and the cap. For this purpose, the corresponding element may, for example, have an edge which runs along an outer circumference of the corresponding element and, when the closure cap is screwed onto the reagent vessel, rests on the edge of the reagent vessel and is pressed against this edge by the closure cap.

In all embodiments of the invention, if necessary, the rows of teeth kreisför mig and concentric with each other. Particularly advantageously, the rows of teeth can run in a common plane and ge in this plane against each other to be movable.

Preferably, the rows of teeth extend in a common plane and are movable in this plane against each other. Preferably, this plane is parallel to a bottom surface of the vessel and / or to a lid surface of the vessel. In this way, half of the rows of teeth can be placed on the bottom of the vessel and the other half of the rows of teeth on a lid of the vessel. So the teeth arranged on the bottom protrude from the Bo into the vessel and the teeth arranged on the lid protrude from the lid into the vessel. Advantageously, the teeth can each have a general cylindrical shape, ie a shape which is defined by suitably shaped side surfaces which extend parallel to a straight line which is perpendicular to the tooth row. In a cross-section perpendicular to this line, the teeth can each have the shape described, with which they cut in one direction and grind in the opposite direction.

Advantageously, the teeth each have a cross section in a plane perpendicular right to its cylinder axis or in a plane parallel to the common Plane in which the rows of teeth extend, which have a sharp corner and a spit zen corner opposite rounded side. The pointed corner and the rounded side are preferably opposite to each other in the direction of movement of the rows of teeth. The teeth thus have preference, in their cross section, a teardrop shape. Advantageously, the spit zen corners of teeth adjacent rows of teeth are opposite and thus the round corners.

In an advantageous embodiment of the invention, those edges of the cross section of the teeth, the verbin the rounded with the rounded side, in the direction of a line along which the row of teeth ver runs, to which the corresponding tooth belongs. This means that the corresponding sides are curved in the same direction as the line along which the row of teeth runs. In an advantageous embodiment, the radius of curvature of one of these two sides can be substantially equal to the radius of curvature of the line along which the row of teeth runs ver.

The drop shape can be advantageously understood as a parallelogram in which one of the two ge separated by a short diagonal of the parallelogram triangles is replaced by a circular arc adjacent to the obtuse angles of the parallelogram to the other of the two triangles. The circular arc here forms the rounded side and the remaining tip angle forms the sharp corner. Advantageously, the drop shape is then mirror-symmetrical around the long diagonal of the original parallelogram, ie the axis of the tip to the farthest point on the round corner. The radius of the circular arc can lie on the short diagonal of the original triangle. Optionally, the arc may be just a half-circle so that the center of the arc lies on the short diagonal of the original triangle. Alternatively, the circular arc can also adjoin the straight sides in such a way that the straight sides lie tangentially on the circular arc. In this case, the midpoint is a bit farther from the top.

Optionally, the drop shape described above may also be bent along the long diagonal of the parallelogram. The mirror symmetry works then lost and the shape is more like a fin.

Advantageously, the vessel can close a circular cylindrical volume. The rows of teeth can then rotate the cylinder axis coaxially.

Advantageously, the cylindrical volume limiting Be tenwand of the vessel have one or more projections in the direction of the tooth rows, wherein the side wall of the adjacent teeth row of this side wall has such a distance that the teeth of this to nearest row of teeth in cooperation with the projections a dissociation cause a portion of the tissue sample. Advantageously, be the projections with the teeth of them lying first Zahnrei hey rubbing a portion of the tissue sample. The projections may, for example, have a triangular shape in a sectional plane perpendicular to the cylindrical axis of the vessel.

In an advantageous embodiment, the vessel and / or the lid may have a base, which is circulated by the rows of teeth. The base may have one or more projections in the direction of the rows of teeth, wherein the basis of the base next row of teeth from this base has a sol chen distance that the teeth of this first row of teeth in egg nem distance range at its minimum distance to the projections, a dissociation of a part effect the tissue sample. Advantageously, the vessel is designed here so that it encloses a cylindrical inner space. The base can also be configured cylindrically with a cylinder axis coaxial with the cylinder axis of the vessel. The rows of teeth can then circulate the base circularly with the cylinder axis as center point.

In an advantageous embodiment of the invention, a diameter of the vessel in the plane in which the rows of teeth extend, greater than or equal to 0.5 cm, preferably greater than or equal to 1 cm, preferably greater than or equal to 2 cm and / or less than or equal to 4 cm , preferably less than or equal to 3 cm.

Preferably, a minimum distance of teeth of adjacent teeth hen in the state of greatest approximation each greater than or equal to 50 miti, preferably greater than or equal to 100 miti, preferably greater than or equal to 200 miti and / or less than or equal to 500 miti, preferably less than or equal to 400 miti, preferably less than or equal to 300 miti. The minimum distance to be described here is to be understood to mean that, when the teeth of adjacent and interacting rows of teeth move together, the teeth move towards each other, then reach the minimum distance and then move away from each other. The minimum distance should be the distance between two points of the profiles of the teeth moving past each other, which has the lowest value.

In an advantageous embodiment of the invention, the housing may have a bottom and a lid, wherein one half of the rows of teeth on the Bo can be arranged and the other half of the rows of teeth can be arranged on the lid. In this case, advantageously arranged on the cover rows of teeth can each be adjacent to the floor arranged rows of teeth in a plane in which the rows of teeth, adjacent. Particularly preferably, a row of teeth arranged on the cover cooperates with a row of teeth arranged on the bottom for tissue dissociation.

According to the invention, adjacent rows of teeth can be moved relative to one another. For this purpose, one half of the rows of teeth relative to the housing can be fixedly arranged and the other half of the rows of teeth, which in each case interact with the fixed rows of teeth for dissociation, can be movable relative to the housing. In an advantageous embodiment of the inven tion, the movable rows of teeth can be arranged on the lid. For this purpose, the lid can advantageously be movable. Alternatively, the cover may be fixed relative to the housing, and the rows of teeth may be designed to be movable relative to the cover and thus also relative to the housing. For this purpose, a suitable mechanism can be integrated in the cover, which FITS preferred from outside the housing when it is closed by the lid, can be driven.

In an advantageous embodiment of the invention, said Ver closing cap of the reagent vessel in its center an opening aufwei sen. It can also advantageously the lid of the vessel an opening tion, which is coaxial with the opening in the closure cap, so that the opening of the closure cap and the vessel are continuous. It Kings then nen the openings in the cap and in the lid be designed so that a drive shaft the lid of the dissociation or the vessel for moving the teeth of the at least one second row of teeth or arranged on the lid at least one row of teeth relative to the teeth of at least a first row of teeth or the at least one row of teeth arranged on the floor can drive. The drive shaft can intervene through the opening in the cap into the opening in the lid of the vessel.

In an advantageous embodiment of the invention, the cover of the Ge housing or dissociation work also a magnetic element aufwei sen, by means of which the at least one row of teeth is driven by a magnet. In this case, an opening in the cap is not required, but may still be provided, for example, to bring substances into the housing can.

Advantageously, the opening in the cap and / or the opening in the lid of the vessel may have a septum, through which liquid keit in the dissociation, the vessel and / or the reagent vessel can be introduced. Advantageously, such a septum can be accommodated at an end of the opening facing the reagent vessel in the lid of the vessel, so that a drive shaft can engage through the closure cap and the opening of the lid into the opening in the lid without being prevented by the septum. As described above, the closure cap and the cover each have an opening, so it is particularly advantageous if the opening in the closure cap is sealed against the opening in the lid.

In an advantageous embodiment of the invention, the cell strainer and / or the bottom of the vessel may comprise elements by means of which a relative movement of the soil relative to the cell strainer, for. B. when driving the lid with means of a motor can be prevented.

It should be noted that according to the invention, the vessel is liquid-tight can be. However, it is also possible that the vessel for liquids, such as the cell suspension is permeable. This is particularly advantageous if the device according to the invention has a reagent vessel and / or a cell sieve as described above. It is then possible first to make a tissue dissociation by means of the dissociation unit or the grinder and then to further process the same in the reagent vessel and / or in the cell sieve, for example to centrifuge or to clean. After the digestion process has ended, the cell suspension can be separated from tissue residues, for example by centrifugation become. It can then be collected in the reagent vessel, which may for example be a centrifuge tube, the cell suspension. It is advantageous here that from the introduction of the tissue sample on the digestion to further processing and examinations, the system can remain closed, whereby the risk of contamination can be significantly reduced while user friendliness is increased.

The said rows of teeth can be firmly integrated in the lid and in the floor. But it is also possible to design the rows of teeth in the lid and / or in the bottom of the vessel as exchangeable inserts. In particular, such a replaceable insert in the lid relative to the lid can be movably ge outsourced so that it is driven to move the teeth. The grinder can thus have, for example, two inserts with one or more rows of teeth, which mesh and rotate in opposite directions.

In an advantageous embodiment of the invention, the dissociation can also be formed onswerk that an insert with at least one row of teeth on the cap of the reagent vessel is arranged and an insert is inserted with at least one row of teeth in the cell sieve or is firmly integrated into this. In this embodiment of the invention, the two inserts can be seen together as a housing in the sense described above or form such a housing.

Advantageously, one of the inserts may have a concentric with the rows of teeth surrounding the rows of sheath, which can act as the outer wall of the housing Ge. It can then be placed with the arranged on the bottom of the housing rows of teeth on the lid of the other insert. The floor can then rest on an edge of said lateral surface and advantageously be slidably mounted with respect to this.

In an advantageous embodiment of the invention, therefore, the vessel for receiving the tissue sample on the one hand have the lid with the at least one second row of teeth and on the other hand the floor with the at least one first row of teeth. The at least one second row of teeth can thus be arranged on an end face of the lid and a lateral surface can rotate around the end face at its edge. Particularly advantageous is the man telfläche a cylindrical surface which is concentric with an axis about which extend the teeth of at least one second row of teeth. The floor may then have a plate, which may also have openings on which the at least one first row of teeth is arranged. In assembled to stand the plate of the bottom can rest on that edge of the lateral surface of the lid, which is opposite to the end face of the lid.

The vessel thus formed may advantageously be arranged at least partially in the closure cap of the reagent vessel and more preferably be screwed with the cap in the opening of the reagent vessel.

According to the invention, a method for the dissociation of tissue is also described which can be carried out with a device as described above. In this case, a tissue sample is introduced into the corresponding device and then the dissociation unit or the grinder is driven. Advantageously, therefore, if necessary, the at least two mutually movable against each other rows of teeth are moved against each other.

If the device is designed so that the tissue sample is cut in one direction and grated in the other, in the inventions to the invention method advantageously the rows of teeth are moved against each other both in the cutting direction and in the rubbing direction.

Advantageously, the rows of teeth with a rotational speed of greater than or equal to 60 U / min, preferably greater than or equal to 200 U / min, preferably greater than or equal to 300 U / min and / or less than or equal to 700 U / min, preferably less than or equal to 500 U / min, more preferred less than or equal 400 rev / min are moved against each other.

In an advantageous embodiment of the invention, the rows of teeth, while they are moved against each other, are moved periodically in the direction of this movement perpendicular direction. The periodic movement can thus take place in the direction of the cylinder axis of the vessel or the axis of rotation of the rows of teeth.

Preferably, the tissue is provided with a liquid such as a medium or enzyme solution. By rotating in both directions and optionally a linear up and down movement of the grinding cylinder, the tissue can be processed.

Optionally, the grinder can also be decoupled from the lid or housing be driven, for example by a magnetic drive. In this case, the plate carrying a part of the teeth can be moved with an external magnet. The corresponding plate can for this purpose have magnetic material.

Depending on the type of tissue, the comminution process can advantageously be carried out between 30 seconds and 10 minutes with one or more direction changes. Preferably, the rotation time> 1 min, more preferably> 4 min and / or <10 min, preferably <7 min betra conditions.

In an advantageous embodiment of the invention may be provided in the device a plurality of vessels with a plurality of grinders. In particular, the vessels may be cavities of a microtiter plate. In this way tissue dissociation can be parallelized. It is also possible to use the cavities in the microtiter plate individually, partially or in parallel. For partial use, for example, disposable cavity strips are suitable, where the base plate serves only as a frame. The lid can be used, for example, as a carrier for a grinding cylinder or as a carrier for several grinding cylinders, e.g. also in the form of stripes, serve.

In an advantageous embodiment of the method can by driving the dissociation plant or the grinder generated cell suspension after driving the dissociation unit or the grinder be cleaned and / or centrifuged. Here, for example, the device may first be placed on a drive device through which the dissociation unit or grinder is driven for tissue dissociation, and then the device may be placed in a centrifuge to centrifuge the dissociated cell suspension in the reagent vessel.

According to the invention, the industrial sample before, during and / or after driving the dissociation unit or the grinder at least one medi um added. This can be done, for example, by a described as described before ben ausgestaltetes septum in an opening of the lid and / or the United closure cap.

In an advantageous embodiment of the invention, a rotation Rich tion and / or a rotational speed of driving the Dissoziati onswerkes or the grinder based on stored during the driving he parameters are stored. These parameters may include, for example, one or more selected from a torque, a rheological parameter and / or an optical density. In this way, the progress of the dissociation can be detected during the course of the execution of the method, so that the tissue digestion can be executed up to a predetermined point, and when predetermined target values of the parameters considered can be ended.

In an advantageous embodiment, a transmitted to the dissociation plant or the grinder by a drive force or zizerswerk on the dissociation transmitted by a drive torque can be measured by means of one or more sensors. From the force thus measured or the torque thus measured, a degree of dissociation of the fabric can be determined. Advantageously, can be checked by driving the Dissoziati onswerkes or the grinder by means of measurement results, which are determined by one or a combination of sensors. In this way, it is possible in particular to set the transit time for optimizing the degree of dissociation. le

In the following, the invention will be explained by way of example of some figures. The features described in the figures can be realized inde pendent of the specific example and be combinatorial under the examples. Like reference numerals denote the same or corresponding features.

It shows

FIG. 1 shows an example of a device according to the invention for the dissociation of tissue,

FIG. 2 shows a further example of a device according to the invention for the dissociation of tissue,

FIG. 3 shows a multiplicity of devices according to the invention for dissociating tissue arranged in a microtiter plate,

FIG. 4 shows a device according to the invention for the dissociation of tissue with a dissociation plant and a pot-shaped cell sieve,

Figure 5 is a device shown in Figure 4 with a reagent vessel, and

Figure 6 shows the device shown in Figure 5 mounted on a device Antriebsvorrich for driving the Dissoziationswerks.

FIG. 1 shows an example of a device according to the invention for the dissociation of tissue. In the example shown, the device 1 for the dissociation of tissue is circular. The device has a base 2, of which in the plan view of Figure 1, a circular side wall 2a can be seen. The bottom 2 is closed by a cover 3, of which in the sectional view of Figure 1, a cylindrical side wall 3 can be seen. The elements 2 and 3 together form a vessel 4 for receiving a tissue sample. The vessel 4 is limited here by the side wall 3 of the lid. The bottom 2 has around a center of the housing 2, a base member 2b, which has an opening in its interior. In this opening can be a centra engage the element of the lid 3.

In the example shown in FIG. 1, the device has a grinder with two mutually movable rows of teeth 5a-5f and 6a-6f. Each of the rows of teeth has a plurality of teeth 5a - 5f and 6a - 6f. The Zahnrei hen are arranged at such a distance from each other that the teeth 5a

- 5f and 6a - 6f of these respective adjacent rows of teeth in a distance range around their minimum distance from one another cause a dissociation of a portion of the tissue sample. The partial images of Figure 1 show the grinder in a state in which the teeth 5a and 6a to 5f and 6f each have a mini malen distance to each other.

In the example shown in FIG. 1, the teeth have a teardrop shape in a cross section perpendicular to the axis of rotation about which the rows of teeth revolve. In the direction of a line along which the teeth 5a - 5f and 6a - 6f are arranged in the ent speaking row of teeth, the drop shape has on the one hand a tip and on the other hand opposite a rounding. The tips of the teeth 5a - 5f of one row of teeth are opposite to the tips of the teeth 6a - 6f of the other row of teeth. Likewise, the curves of these teeth are opposed to each other. As a result, the teeth 5a-5f, together with the teeth 6a-6f, cut the tissue as they move in the direction of the tips and tear the tissue as they move in the direction of the curves.

In order to improve the dissociation result, the wall of the lid 3 has triangular projections 7a-7d projecting from the wall in the direction of the teeth 5a

- 5f and 6a - 6f and in the direction of the center of the vessel 4 hervorra conditions. Move the teeth 6a - 6f past these projections 7a - 7d, tissue between the projections 7a - 7d and the teeth 6a - 6f rubbed zer.

Similarly, the base member 2b has triangular teeth 8a-8c projecting from the base 2b in the direction of the teeth 5a-5f and 6a-6f. When the teeth 5a-5f of the inner row of teeth move past these projections 8a-8c, between the teeth 5a-5f and the projections 8a

- 8c tissue rubbed. The teeth 6a-6f of the inner row of teeth can be arranged on the lid 3 and moved with it. The teeth 5a-5f of the outer row of teeth are advantageously arranged on the bottom 2 of the vessel 4 and fixed with respect to this. Accordingly, the base element 2b with the projections 8a-8c is advantageously arranged on the underside of the vessel 4 and fixed against it. If now the cover 3 or an element arranged on the cover 3 and carrying the teeth 6a-6f are rotated, the teeth 6a-6f move in a circle past the teeth 5a-5f and the projections 8a-8c and cause a dissociation of the teeth tissue. At the same time, the projections 7a-7d on the cover move past the teeth 5a-5f, causing dissociation of the tissue there.

FIG. 2 shows a further embodiment of a device according to the invention for the dissociation of tissue. The construction of the device shown in Figure 2 corresponds to that shown in Figure 1, so that reference should be made to the description of Figure 1. In the following, only the differences from FIG. 1 are executed.

In FIG. 2, the teeth 5a-5f and 6a-6f likewise have a teardrop shape which, however, is bent in the direction of the center point and the axis of rotation with respect to the teardrop shape shown in FIG. As a result, these teeth 5a-5f and 6a-6f likewise have a pointed side and a round side, which, however, in FIG. 2, unlike in FIG. 1, are connected by curved sides. In the example shown, the inside of the outer teeth 5a-5f is optionally parallel to the course of this row of teeth curved, that is, has substantially the same radius of curvature. The outer sides of the teeth 5a - 5f and 6a - 6f are optionally curved with a smaller radius of curvature than the toothed row.

FIG. 3 shows a multiplicity of devices according to the invention, which are arranged in 4x6 cavities 10 of a microtiter plate 9. For each of the cavities 10, a cover 3 is provided which carries the teeth 6a, 6b of one of the rows of teeth. Above each cavity 10 forms a vessel 2, in which teeth 5a, 5b, 5c of a white direct row of teeth are arranged. The arrangement of the teeth and Ausgestal direction of the teeth can be realized as shown in Figures 1 and 2. The Teeth 5a, 5b, 5c can also be designed as inserts for the cavities 10, so that they can be used in commercially available microtiter plates 9.

Tissue processing can be done in the examples of Figures 1 to 3 in example as follows. There may be 24 cavities as shown in FIG. An inner diameter of the cavities can be, for example, 2 cm. The teeth 5a-5f can be rotated with respect to the teeth 6a-6f, for example at a rotational speed of 60-700 rpm. For other scaling of the cavities and the grinder, the rotation speed can be adjusted accordingly. The duration of the dissociation can be chosen, for example, between 1 and 10 min. One or more changes of direction can be made during this time. In this way, for example, pieces of tissue can be dissociated from a mass of 50 mg to 3 g. A minimum distance of the teeth of adjacent rows of teeth, for example betra between 50 and 500 pm.

FIG. 4 shows an exploded view of an apparatus according to the invention for the dissociation of tissue with a dissociation unit 1, which is configured here with a lid 1a and a bottom 1b. The dissociation plant is in Figure 4 at least partially inside a cap 41 of a Rea genzgefäßes, which is shown in Fig. 5, arranged. The device also has a pot-shaped cell sieve 42, in which the dissociation unit 1 is arranged. The cap 41, the lid la, the bottom lb and the cell strainer 42 are cylindrical in Figure 4 and arranged with coaxial cylinder axes. In Figure 4, the components are shown exploded in the direction of the cylinder axis, in the assembled state, however, they are all in one another.

The cell strainer 42 is cup-shaped in the example shown, in the form of a closed on one side circular cylinder. The cell screen 42 has four cell screen surfaces 44 in the cylinder surface. Moreover, the dissociation plant 1 facing away from the end face 43 of the cell strainer 45 is closed by a cell. Those of the cell screen surface 45 opposite end face of the cup-shaped cell strainer is open. The dissociation plant has on the one hand the lid la and on the other the bottom lb. In this case, the lid 1a has an end face, which is not visible in the figure, on which a plurality of teeth 5a, 5b are arranged so that they are parallel to the cylinder axis of the lid. The teeth of the cover la are surrounded by a lateral surface lc, which runs along an edge of the front side of the lid, on which the teeth 5a, 5b are arranged. The lateral surface lc here is a cylindrical surface whose cylinder axis coincides with the Zy cylinder axis of the dissociation unit 1 as well as the cell strainer 42. The bottom 1b of the dissociation plant 1 has teeth 6a which cooperate with the teeth of the cover for dissociation.

In the example shown la, the cover la also has a central through-opening ld, which extends in the middle of the unrecognizable Stirnflä surface and is surrounded by the teeth 5a, 5b. This opening ld he leaves on the one hand, a coupling of a motor for driving the lid la and beyond also the introduction of liquids in the dissociation onswerk 1. Advantageously, while the opening at its cell sieve 42 facing the end having a septum. The opening ld can be configured as a tubular channel extending from the end face of the lid la to the end of the lateral surface lc of the lid.

Advantageously, the closure cap 41 also has an opening which is coaxial with the opening 1d and which is sealed off from the opening 1d.

FIG. 5 shows the dissociation plant shown in FIG. 4 as an exploded view with a reagent vessel 51. As in FIG. 4, the dissociation plant 1 is arranged in the closure cap 41 with a lid 1a and a bottom 1b and surrounded by the cell strainer 42. With regard to these elements, reference is made to the description of FIG. 4.

The reagent vessel 51 has a circular cylindrical shape with a tapered ver closed end and an opening 52 in that of the tip opposite the end face of the cylindrical shape. The closure cap 41 can be screwed onto the opening 52 and surrounds or surrounds the opening 52. The dissociation unit 1 and the cell sieve 42 are in the screwed state in the closed position. cap 41 and disposed in the opening 52 or just behind the opening 52 in the interior of the reagent vessel 51. FIG. 5A shows the device in a perspective view. Figure 5B shows the device in conjunction with a drive device 53 as a perspective view and Figure 5C shows the device shown in Figure 5B assembled in a sectional view.

In Figure 5C it can be seen that the cap 41 is screwed or applied to the reagent vessel 51 while the cell strainer 42 between egg nem edge around the opening 52 of the Reagenzgefäßes 51 and the Verschlusskap 41 clamps. As a result, the cell screen 42 is held in the opening 52 in the interior of the reagent vessel 51 immediately behind the opening 52. A shaft 54 of the drive device 53 engages in the lid la of the Dissoziati onswerks, whereby this is rotatable. The bottom 1b of the dissociation plant 1 is fixed relative to the cell strainer and the reagent vessel 51 and is not rotated when the lid 1a is rotated. As a result, the teeth 5a of the cover la move relative to the teeth 6a of the bottom 1b and thus cause a dissociation of tissue introduced into the dissociation mechanism 1. The lid 1a and the bottom 1b of the dissociation unit 1 form a housing within which the tissue is dissociated. In the example shown, however, the housing is not fluid-tight so that the cell suspension can flow out of the dissociation apparatus 1 when the apparatus is rotated so that the reagent vessel 51 faces downwards or when the apparatus is placed in a centrifuge in which the centrifugal force is applied in the direction of Rea genzgefäßes 51 acts.

The reagent vessel 51 may advantageously be a standard laboratory vessel, and also the lid 41 may be a standard laboratory vessel lid. The lid can also be fixed by means of a circumferential clamping ring.

The implementation of the coupling of the drive 53 to the bottom lb of the dis sociators 1 can be fitted with plain bearings to ensure a low-resistance rotation and simultaneous tightness.

FIG. 6 shows the device shown in FIG. 5, applied to a tissue grinder device, which acts here as a drive device 53. The tissue grinder Device 53 may in this case again have a shaft 54 which engages in the lid 1a of the dissociation unit 1 and can rotate it.

In the following, an expiry of a method according to the invention will be described by way of example, in which the device described can be used.

In a first step of tissue processing, a tissue sample to be examined is introduced into the dissociation unit 1 or the housing of the dissociation unit 1. For the lid la with the tissue and a liquid, for. As medium or enzyme solution, are filled and merged with the Bo lb in the cell strainer 42 via a union nut or cap 41. By rotation, preferably in both directions, and optionally linear up-down movement of the dissociation unit 1, the tissue is now processed by cutting and / or grinding operations. Optionally, the cutting unit can also be operated decoupled from the lid la, z. B. by an integrated magnet. In this case, the lid 1a and the dissociation unit 1 can be decoupled and the lid 1a can be driven by rotation of a magnet over the lid.

During the digestion process, the lid la of the cutter is connected to the drive 53 and is driven by it. The bottom 1b of the dissociation unit 1 or the housing 1 is fixed here and can be inserted directly into the cell sieve 42. In order to produce a closed system, the bottom 1b of the dissociation unit 1, which may be integrated in the cell strainer 42, is screwed to the lid 1a of the dissociation unit 1 through the cap 41 on the reagent container 51.

After completion of the digestion process, the cell suspension example, by centrifugation of tissue residues is separated. Further washing steps to increase the cell yield can be effected by a septum, which can be integrated into the opening 1 d in the lid lb. The cell suspension can be collected in the reagent vessel 51 and can thus remain in a closed system from the introduction of the tissue sample via digestion to further investigations, which considerably reduces the risk of contamination and at the same time simplifies the user-friendly operation. increased.

Tissue processing with the described rows of teeth can be used for centrifugal tubes 51 and cell filters (with, for example, a mesh between 10 and 200 microns) at a Schneidwerkinnendurchmes of for example 2 cm and a rotational speed of for example 10 to 700 U / min. respectively. Other scalings are possible, in which case the rotation speed is preferably adjusted. Depending on the type of tissue, a rotation time of, for example, 1 to 10 minutes and one or more directional changes of the rotation may be useful. With a suitable adaptation of the size of the housing, dissociation of tissue pieces from a mass of, for example, 50 mg to 3 g can be achieved. The minimum distance between teeth of adjacent rows of teeth is preferably between 50 and 500 pm.

A reagent vessel 51 is suitable for the processing of one or more tissue samples simultaneously. Advantageously, you can lines, z. B. be provided in the form of plain bearings to ensure a resistance poorer rotation and simultaneous tightness. In this case, in particular, the opening in the closure cap 41 and in the lid la can be sealed against one another. By providing the cell filter 42, tissue remnants of single cells in the closed system can be separated.

It is particularly advantageous if the degree of digestion, ie the dissociation efficiency, takes place via an online process control. In this case, the digestion of the tissue sample by means of sensors, for. B. based on a turbidity measurement, via an optical sensor and / or a torque measurement and / or rheological measurements of the digestion tools. The recorded parameters can be analyzed and converted into a generally valid differential equation for the description of the viability and yield of the Zielzel len at different process parameters. Based on the online process control, an intelligent control of the rotational direction and speed of rotation of the grinder as well as the duration of the tissue dissociation can take place. This allows each tissue sample to be individually sized to ensure optimal cell yield. The method according to the invention can be carried out, for example, by first loading the dissociation unit 1 with a tissue sample and determining the weight of the tissue sample. Medium may also be added. For dissociation, the rotational direction, rotational speed and / or duration of the dissociation can be controlled by the on-line acquisition of process parameters such as torque, and rheological parameters and optical density. The recorded parameters can be analyzed and forwarded by means of decay equations to a feedback system of the motor control of the drive 53. The feedback system can allow individual dissociation of each tissue sample to increase efficiency and yield.

Purification of the generated cell suspension is possible. For this purpose, the reagent vessel 51 can be placed in a centrifuge. After a first centrifugation as many washing steps, for example, by he neute addition of medium via the integrated septum in the engine coupling done. The purified cells then collect at the bottom of the Rea genzgefäßes 51st

The device according to the invention and the method according to the invention he allow a very targeted dissociation of tissue, with a desired final state of dissociation can be achieved very precisely. By a suitable combination of tooth geometries, cell sieves and dissociation units if required, and intelligent control, predetermined dissociation levels can be exactly adjusted. The described combination of tissue dissociation and centrifugation in a closed system allows complete processing of a tissue sample without the risk of contamination. The described drop shapes and curved shapes of the teeth, the tissue can be cut very gently and / or ge grind, which can be selected between cutting and grinding as needed. By controlling the direction of rotation, the predominant de force can be determined. In comparison to processes according to the prior art, tissue samples are not only processed more gently for the cells, but also save time. The tissue no longer needs to be cut into very small pieces with a scalpel, but can be processed in bulk or in a few pieces. By integrating a Cell sieve, the tissue sample can be purified directly without further pipetting steps. The process is semi-automatable, which gives a higher reproducibility and standardization. The technology can be inserted into automated processes.

Another advantage of the system is its parallelizability. The possibility of simultaneous use of several centrifugal tubes, the tissue processing can be parallelized so that several samples can be processed simultaneously. The advantage on the one hand saves time and on the other sources of error are minimized. When processing several samples, all necessary preparatory work steps for all samples can be carried out simultaneously and do not need to interfere anymore. An improved comparability of the results is given. An example of this is a certain incubation time with an enzyme solution, which can only be maintained if samples are added exactly the same time before the dissociation with the enzyme.

The centrifuge tube format allows the system to be integrated into an automated process. An automation prevents differences in the workup as it would be the case in the implementation by different employees. This leads to an improved reproduction and comparability of the results.

Since the device in its centrifuge tube variant is disposable, the probability of contamination and auxiliary resources can be minimized very much.

The device is suitable for cell disruption as well as for the generation of single cells from tissue.

Cell disruption is e.g. protein isolation or nucleic acid isolation, as well as other processes involving analysis of cell constituents.

The gentler dissociation of tissue is advantageous when individual cells are needed. This is the case, for example, in the examination of tumor cells by mass spectrometry, infrared spectroscopy or ELISA. Potentiel les tumor tissue can be dissociated for histological examination. In addition, the production of single-cell suspensions is important for the generation of 2D cultures, 3D cultures, single-cell characterization, high-throughput drug screening, and organ-on-chip technology. The system is not limited to application to tumor tissue, but can be theoretically adapted to any type of tissue.

Claims

claims

A device for dissociation of tissue, comprising

at least one dissociation work for receiving a tissue sample, at least one cup-shaped cell sieve,

wherein the at least one Dissoziationswerk is at least partially disposed in the pot-shaped cell strainer.

2. Device according to one of the preceding claims,

the dissociation work

at least one vessel for receiving the tissue sample,

and a grinder arranged in the vessel,

wherein the grinder has at least a first row of teeth with a plurality of teeth and at least one second row of teeth with a plurality of teeth,

wherein the at least one first of the rows of teeth is arranged on a bottom of the vessel and the at least one second of the rows of teeth are arranged on a lid of the vessel,

wherein the at least one first row of teeth is movable relative to the at least one second row of teeth, so that the teeth of the first row of teeth are movable past the teeth of the second row of teeth, wherein in each case one of the at least one first row of teeth and one of the at least one second row of teeth at such a distance are arranged adjacent to each other, that the teeth of these respective adjacent rows of teeth, when they are moved past each other in a distance range around their minimum distance from each other ei ne dissociation of a portion of the tissue sample cause.

3. Device according to the preceding claim,

wherein the teeth are configured to intersect the portion of the tissue sample as they pass one another in a direction and as they oppose each other in that direction Move towards each other, rub the part of the tissue sample.

4. Device according to one of the preceding claims,

further comprising a reagent vessel,

wherein the cell strainer and the Dissoziationswerk are angordnet in an opening of the Re agenzgefäßes.

5. Device according to one of claims 2 to 4, wherein the rows of teeth arranged in the lid of the vessel are movable relative to the reagent vessel and arranged in the bottom of the vessel Zahnrei hen are fixed to the reagent vessel.

6. An apparatus for dissociation of tissue, comprising

at least one vessel for receiving a tissue sample,

and a grinder arranged in the vessel,

wherein the grinder has at least two mutually movable tooth rows,

wherein each of the rows of teeth has a plurality of teeth, wherein adjacent ones of the rows of teeth are spaced apart such that the teeth of each adjacent row of teeth cause dissociation of a portion of the tissue sample within a range of their minimum distance apart are configured to intersect the portion of the tissue sample as they pass each other in one direction and, as they move past each other in an opposite direction to that direction, they rub the portion of the tissue sample.

7. Device according to the preceding claim,

further comprising a reagent vessel,

wherein the vessel and the grinder are angordnet vessel in an opening of the reagent,

wherein the rows of teeth arranged in the lid of the vessel are movable relative to the Reagenzgfäß and in the bottom of the vessel arranged rows of teeth relative to the reagent vessel are fixed.

8. Device according to one of the preceding claims,

wherein the cell strainer and the dissociation mechanism or the vessel and the grinding mechanism are at least partially disposed within a closure cap of the reagent vessel,

wherein the closure cap surrounds the opening of the reagent vessel.

9. Device according to one of claims 2 to 8,

the rows of teeth are circular and concentric with one another.

10. Device according to one of claims 2 to 9,

wherein the rows of teeth extend in a common plane and in this plane are mutually movable.

11. Device according to one of claims 2 to 10,

the teeth each having a cross section parallel to the common plane, having a sharp corner and a rounded corner opposite the acute corner,

wherein the pointed corners of teeth of adjacent rows of teeth are on the opposite.

12. Device according to one of claims 2 to 11,

wherein those edges of the cross section of the teeth connecting the tip with the rounded side, are curved in the direction of a line along which runs the row of teeth to which the corre sponding tooth belongs.

13. Device according to one of claims 2 to 12,

wherein a side wall bordering the vessel has at least one or a plurality of projections in the direction of the rows of teeth, wherein the side wall of the adjacent teeth row of this Be tenwand is arranged at such a distance that the teeth ser first lying row of teeth in a distance range around their minimum distance cause the projections to dissociate part of the tissue sample.

14. Device according to one of claims 2 to 13,

the vessel having a base which will run around the rows of teeth, the base having one or a plurality of projections in the direction of the rows of teeth, wherein the base of the first row of teeth from this base is at such a distance angeord net, that the Teeth of this next row of teeth in a distance range around their minimum distance to the projections ei ne dissociation of a portion of the tissue sample cause.

15. Device according to one of claims 2 to 14,

wherein a diameter of the vessel in the plane in which the rows of teeth are greater than or equal to 0.5 cm, preferably greater than or equal to 1 cm, preferably greater than or equal to 2 cm and / or less than or equal to 4 cm, preferably less than or equal to 3 cm is.

16. Device according to one of claims 2 to 15,

wherein the minimum distance of teeth of adjacent teeth of the rows of teeth is greater than or equal to 50 pm, preferably greater than or equal to 100 pm, preferably greater than or equal to 200 pm and / or less than or equal to 500 pm, preferably less than or equal to 400 pm, preferably less than or equal to 300 pm is.

17. Device according to one of claims 2 to 16,

wherein one half of the rows of teeth on the lid of the vessel is arranged and the other half of the rows of teeth is arranged at the bottom of the vessel is wherein the arranged on the cover rows of teeth each arranged at the bottom rows of teeth in a plane in which the rows of teeth are adjacent.

18. Device according to the preceding claim,

wherein the rows of teeth arranged on the cover are movable relative to the lid.

19. Device according to one of claims 8 to 18,

wherein the closure cap has an opening at its center and the lid of the vessel has an opening for opening the closure. cap has coaxial opening,

wherein the openings in the cap and in the lid are designed so staltet that a drive shaft can drive the cover of the vessel to BeWe movement of the teeth of the at least one second row of teeth relative to the teeth of the at least one first row of teeth.

20. Device according to one of claims 2 to 19,

wherein the lid of the vessel comprises a magnetic element by means of which the lid for movement of the rows of teeth is driven by means of a magnet.

21. Device according to one of claims 8 to 20,

wherein in the opening in the cap and / or in the opening in the lid of the vessel, a septum is arranged, through which liq fluid into the vessel and / or the reagent vessel can be introduced.

22. Device according to one of claims 2 to 21,

wherein the cell strainer and / or the bottom elements, preferably webs, for preventing relative movement of the bottom relative to the cell strainer.

23. Device according to one of claims 19 to 22, wherein the opening in the lid against the opening in the closure cap is sealed.

24. Method for the dissociation of tissue,

wherein a tissue sample is introduced into a device according to any one of claims 1 to 23, and

then the Dissoziationswerk or the grinder is driven.

25. Method according to the preceding claim,

wherein the rows of teeth are circular and with a speed of Rotationsge of greater than or equal to 60 U / min, preferably greater than or equal to 200 U / min, preferably greater than or equal to 300 U / min and / or less than or equal to 700 U / min, preferably smaller or be moved equal to 500 U / min, preferably less than or equal to 400 U / min against each other.

26. The method according to one of the two preceding claims, wherein after driving the dissociation unit or the grinder ei ne generated by driving the dissociation unit or the grinder cell suspension is purified.

27. The method according to any one of claims 24 to 26,

wherein the tissue sample before, during and / or after driving the dissociation mechanism or the grinder at least one medium is added.

28. The method according to any one of claims 24 to 27,

wherein a rotational direction and / or rotational speed of driving the dissociation mechanism or grinder is controlled based on parameters detected during driving, the parameters comprising one or more of torque, rheological parameter and / or optical density.

29. The method according to any one of claims 24 to 28, wherein a force transmitted to the Dissoziationswerk or the grinder force is determined by means of one or more sensors, and from the thus determined force a degree of dissociation of the tissue is determined.

30. The method according to any one of claims 24 to 29,

wherein the rows of teeth, while they are moved against each other, are peri odically moved in the direction of movement against each other perpendicular direction.

31. The method according to any one of claims 24 to 30,

wherein the Zelllsuspension is centrifuged in a reagent vessel, in the opening of the dissociation plant or the grinder is arranged.