DE102008024408A1 - Device for investigation of a migration of adherent cells from a coating of a carrier coated by protein, comprises a barrier, which stands in contact with the carrier and is removable from the carrier - Google Patents

Abstract

The device (10) comprises a barrier (12), which stands in contact with the carrier and is removable from a carrier (25). The barrier prevents a growth of the cells on the carrier in the area of the contacts, so that an area kept free from the cells during the cultivation of cells on the carrier. The removal of the barrier permits the migration of the cells in the area. The barrier consists of a material on the area, which stands in contact with the carrier and the material non adheres on the coated carrier. The device (10) comprises a barrier (12), which stands in contact with the carrier and is removable from a carrier (25). The barrier prevents a growth of the cells on the carrier in the area of the contacts, so that an area kept free from the cells during the cultivation of cells on the carrier. The removal of the barrier permits the migration of the cells in the area. The barrier consists of a material on the area, which stands in contact with the carrier and the material non adheres on the coated carrier. The carrier is the base of the cell culture container or is arranged on the base of the cell culture container. The carrier is a plate made of glass or first plastic. The cellculture container is a Petri dish (11) or a dent of a microtiter plate made of second plastic. The barrier consists of third plastic and/or stainless steel. A means by which the barrier is pressed in the area of the contacts on the carrier is arranged on the barrier, the carrier or the cell culture container. The means is a spring (14) or clasp for clamping the barrier on the carrier or the cell culture container, formed from metals or fourth plastic or is a magnet. The spring and the barrier or the clasp and the barrier are formed in single piece manner. The barrier is produced in injection molding method. The barrier has coating made of further protein on the area, which stands in contact with the carrier. The surface of the barrier is functionalized through physical or chemical modification of the surface or through applying of inert coating. The carrier and the barrier or the cell culture container and the barrier comprise plane surface corresponding in the area of the contacts in its form. The barrier is formed so that the area free from cells has width of 0.5-2 mm or diameter of 0.5-2 mm. The barrier is formed so that the area free from cells is widened on both ends of the strip. An independent claim is included for a method for investigation of a migration of adherent cells from a coating of a carrier coated by protein.

Description

The The invention relates to a device for studying a migration adherent cells on at least one protein existing coating of a carrier. There is one in contact with the wearer and by the wearer removable barrier provided on the support in the area of contact prevents growth of the cells, so that there in a cultivation of the cells on the carrier left by the cells remains area. A removal of the Barrier allows the cells to migrate to the free ones Area inside.

Out Todaro GJ et al., J. Cell. Physiol. 1965, 66 (3): 325-33 is a so-called scratch assay known. In this case, a cell-free strip is produced in a cell lamina that has grown to a single layer on a support until it reaches its confluence by scraping a strip of cells of this cell lawn away with a pipette tip. Subsequently, the surrounding cells can migrate into the vacated area. The migration of the cells can be investigated. However, the method has the disadvantage that cells are destroyed by the scratching. As a result, substances are released from the destroyed cells, which can influence the migration and possibly a factor to be examined with regard to its influence on the migration. Furthermore, scraping away cells from those remaining cells that have been linked to the scraped cells via adhesive contacts can result in permeabilization and thus also in the migration behavior of these cells.

Farther is from the company ibidi GmbH, Am Klopferspitz 19, 82152 Martinsried, Germany announced an insert for a cell culture vessel, with which the migration of cells can be examined. This Cell culture insert is made of silicone. On the bottom, with which cell culture insert on the bottom of a cell culture vessel is set, this is sticky or adhesive. After this Place on the bottom of the cell culture vessel The use adheres there independently and forms two liquid-tight by a partition from each other separate chambers, in which on the bottom of the cell culture vessel adherently growing cells until reaching confluency can be cultivated. By the stickiness of the bottom In use, cell growth under the bulkhead is completed avoided.

To removing the insert from the bottom of the cell culture vessel remain two defined cell areas, which by a cell-free Range of the width of the partition between the two chambers are separated from each other. The migration of cells into the cell-free Area inside can be examined.

task It is the object of the present invention to provide an improved device and to provide an improved method by which migration becomes more adherent Cells on a carrier coated with at least one protein can be examined.

These The object is achieved by the features of claims 1 and 17 solved. Advantageous embodiments of Invention will become apparent from the features of the claims 2 to 16 and 18 to 25.

According to the invention a device for studying a migration more adherent Cells on a coating consisting of at least one protein a carrier provided.

Prefers is the protein an extracellular Matrix protein, in particular a fibronectin or a collagen. at the carrier may be, for example, the bottom of a cell culture vessel act. The carrier may also be on the bottom of a cell culture vessel be arranged. The carrier may for example be a plate or a plate, for example made of glass or a first plastic, his. At the plate or the plate It can be a coverslip or a slide act. Instead of an arrangement on the bottom of a cell culture vessel But it is also conceivable that on the plate or another Carrier adherent growing cells only from a drop of a cell culture medium and the carrier or the plate is placed in a moist chamber, so that the drop can not dry.

According to the invention at least one device in contact with the carrier in the device standing and removable from the carrier barrier provided which on the support in the area of contact growth prevents the cells, so in the area of contact at a Cultivation of the cells on the support one of the cells reserved area remains. The contact can be, for example manufactured and maintained by that barrier pressed on the bottom of the cell culture vessel and is clamped there. To allow the clamping the barrier z. B. be sized so that after insertion into the cell culture vessel through a wall of the Cell culture vessel is clamped. At a Petri dish as a cell culture vessel would be the dimensions of the barrier then on the inner diameter of the Petri dish Voted.

Removal of the barrier allows migration of the cells into the reserved area. According to the invention, it is further provided that the barrier on the region in contact with the carrier consists of a material which does not adhere to the coated carrier.

The Inventors of the present invention have recognized that by the Scrape the cells in the scratch assay one on the support existing coating is removed from protein. This leads to, that the cells in the subsequent migration directly on the material of the carrier, for example plastic or Glass, migrate. This leads to artificial results, which in terms of the native behavior of in contact with a extracellular matrix standing cells little meaningful are.

The Inventors of the present invention have further recognized that with the removal of the cell culture insert of the company ibidi GmbH from the bottom of a cell culture vessel proteins, with which the bottom of the cell culture vessel coated is to be removed. The cause for this is the stickiness the bottom of the cell culture insert. The study of cell migration in that by removing the cell culture insert protein-free area also provides artificial results because The cells are directly on the, for example, glass or plastic migrate existing bottom of the cell culture vessel.

in the Unlike the cell culture use of the company ibidi GmbH sticks the Barrier of the device according to the invention not even stuck to the carrier. It consists at least in that area in contact with the carrier made of a material, which does not adhere to the coated support. The Material is therefore not adhesive, such as soft Silicone, so that when removing the barrier to get the coating remains. The main advantage of the invention Device for use coming barrier is that through maintaining the coating the adherent cells after removing the barrier on the coating and not on the Material from which the carrier is made to migrate. Another advantage of the barrier of the device according to the invention is their reusability because of the properties Do not change the barrier during repeated use. In contrast, the use sold by the company ibidi GmbH only very limited reusable because of its stickiness decreases with the number of uses, so the use is not more on the carrier.

through the device according to the invention makes it possible to study the migration of cells on a defined coating. Cells are in their native environment in contact with the organism an extracellular matrix. This matrix has a decisive one Influence on the migration behavior of the cells. The expressiveness a performed with the device according to the invention Investigation of the migration behavior is therefore much greater as the expressiveness of one with the use of the company ibidi GmbH or by means of a scratch assay. The device according to the invention can also be several, in particular arranged in parallel, containing barriers. That is in particular then advantageous if in the cell culture medium in the migration of Cells a liberated from the cells substance, in particular a to other cells acting as a messenger substance to be examined should. The total number of cells migrating in the device This is greater than the presence only a barrier, so overall a larger one Amount of this substance is released. Contains the device according to the invention several of the barriers, these can, for example, by means a bridge, be connected to each other. This is the handling the barriers are simplified.

at the cell culture vessel is preferred around a Petri dish or a well of a microtiter plate. Both vessels and their coating with protein are established in the laboratory practice. The Petri dish or the well the microtiter plate or the entire microtiter plate can consist of a second plastic. Plastics are particularly good suitable for coating with proteins. Depending on the coating selected protein may be a second plastic with a selected for the protein opposite charge be used to promote the binding of the protein to the plastic.

The Barrier preferably consists of one, in particular solid, biocompatible Material, preferably a third plastic and / or a, preferably inert, metal. The metal may be stainless steel act. A biocompatible material is understood as meaning a material which by its nature the growth and / or the multiplication not or not significantly affected by cells. An influence For example, the growth and / or proliferation of cells may be caused by substances that releases the plastic. The Barrier can also be embedded in the third plastic metal contain. As a result, for example, a magnetic force the barrier will be exercised when it is at the metal around a magnetic or magnet attractable metal is. At the same time, advantageous properties of the third plastic, such as corrosion resistance or antiadhesive Properties to be used.

Particularly preferably, the third plastic is, in particular high molar gular, polyethylene (PE), polystyrene (PS), polypropylene (PP), teflon, polymethyl methacrylate (PMMA), acrylonitrile-butadiene-styrene copolymer (ABS), polyethylene terephthalate (PET), polyphenylene ether (PPE), styrene / acrylonitrile ( SAN), styrene / butadiene (SB) or any other plastic used in medicine or biomedicine.

at a preferred embodiment, in particular at the barrier, the carrier or cell culture vessel, at least one means provided by means of which the barrier in the area of the contact is pressed onto the carrier. This can more reliably prevent the Cells infiltrate the barrier in the area of the contact. In which Means can be one at the barrier, the carrier or the cell culture vessel, in particular from the or another metal or the third or a fourth Plastic formed, spring or clip for clamping the barrier the carrier or cell culture vessel or also act on a magnet. Is that included in the barrier or the barrier forming metal magnetically or by a magnet dressable, such. As stainless steel, the agent can under the carrier be arranged magnet through which the barrier to the carrier is pressed. The magnet can be a permanent magnet or act an electromagnet. To a predefined arrangement of Magnet in relation to the carrier and / or a Easy to use, the magnet can turn on one in its shape to the shape of the carrier or cell culture vessel be arranged coordinated device. At the facility it can z. B. act to a lid of a Petri dish, at the Outside the magnet is fixed. To provide a This lid is then carried along with the magnetic field acting on the barrier arranged upwards and the Petri dish put into the lid.

The Spring may be formed, for example, as a wire hanger. In each case a wire hanger can be on opposite Arranged sides of the barrier and be sized to the barrier thus on the walls or the wall of the cell culture vessel can be clamped to the barrier thereby to the ground of the cell culture vessel. The wire hanger have the advantage that they can be bent so that they adapt to petri dishes or other cell culture vessels allow with different diameters or dimensions. Preferably, the spring and the barrier or the clasp and the barrier formed integrally. The barrier can be prepared for example in an injection molding process. at one-piece design of spring and barrier or clasp and barrier is an injection molding process in one single process step possible. The barrier can thereby produced inexpensively in large quantities become.

Preferably has the barrier, especially on the one with the carrier in contact area, no coating from or another Protein up. It is particularly preferred if the barrier, in particular at the area in contact with the carrier, so is that they have no coating from one or more Protein. Even if such a barrier in one Solution for the coating of plastics per se suitable protein, this protein does not remain adhere to the barrier. This can be avoided very reliably that the cells are on a coating of the barrier, approximately out of extracellular matrix proteins formed by the cells themselves, adhere. This can also be avoided when removing the Barrier injured cells adjacent to these adherent cells and possibly cause migration signals become.

The Surface of the barrier, especially the surface at the area in contact with the carrier, preferably by a physical or chemical modification the surface or by applying a, preferably inert coating, be functionalized. This can be an adhesion be avoided and / or achieved on the coated carrier be that the barrier is such that it does not coat out or another protein. The surface For example, it may be functionalized by plasma coating. The surface can also by means of a silane, in particular a functional organosilane, be functionalized. This kind of Functionalization is also called silanization.

Prefers have the carrier and the barrier or the cell culture vessel and the barrier in the area of the contact is corresponding in shape, in particular plane, surfaces on, leaving in the area of contact the smallest possible space between the barrier and the carrier remains. Preferably, the barrier is shaped such that the area kept clear of the cells, in particular 0.1 to 3 mm, preferably 0.5 to 2 mm wider, strips or a, in particular 0.1 to 3 mm, preferably 0.5 to 2 mm diameter, Circle is. Time can then be easily analyzed for migration be determined until the strip or the circle is overgrown. Alternatively, at different times or continuously the speed with which the cells are kept in the free Grow in area.

Preferably, the barrier is shaped such that the area kept clear of the cells is widened at both ends of the strip. This will ensures that the barrier can be fixed more stably and reliably with regard to preventing the migration of the cells in the region of the contact on the support and that a planar surface of the barrier rests flat on the support.

• a) Aussäen der Zellen auf dem Träger in einem Zellkulturmedium,
• b) Kultivieren der Zellen bis sie in einem die Barriere umgebenden Bereich konfluent sind,
• c) Entfernen der Barriere und Erfassen des Ausmaßes des von den Zellen freigehaltenen Bereichs und
• d) Weiterkultivieren der Zellen und Erfassen einer Veränderung des Ausmaßes des von den Zellen freigehaltenen Bereichs.

Furthermore, the invention relates to a method for investigating a migration of adherent cells by means of an inventions to the invention device. The method according to the invention comprises the following steps:

• a) sowing the cells on the support in a cell culture medium,
• b) culturing the cells until confluent in an area surrounding the barrier,
• c) removing the barrier and detecting the extent of the area vacated by the cells and
• d) further culturing the cells and detecting a change in the extent of the cell-free area.

The Detecting the extent of the cells kept free from the cells Area and its change can by means of, in particular regular intervals photographic recordings or by means, in particular in real time successful, video microscopy done. The migration of the cells can be analyzed. The analysis of the migration can be the analysis a migration rate, a migration direction, a protein expression, a secretion of a messenger and / or a protein localization include. The messenger substance is a substance that for the transmission of signals or information between Cells serves. The analysis of protein expression and / or protein localization can be done immunocytochemically. In an immunocytochemical analysis it is a well-known method in which certain proteins of a cell specifically by means of antibodies or antibody fragments are detected. The antibodies or Antibody fragments can themselves be a labeling substance or by means of against these antibodies or antibody fragments directed and a marker substance having secondary antibody or antibody fragments are detected. The detec tion The protein is then detected by detecting the presence of the protein Labeling substance. The marking substance may be a Fluorescent dye, a detectable by a specific reaction Enzyme, such as peroxidase or alkaline phosphatase, or act a molecule, which specifically with high affinity specific another molecule binds, such. Biotin, avidin, streptavidin or phalloidin. To enable a particularly sensitive detection, may also be antibodies or antibody fragments be used, which is a radioactive isotope as a marker exhibit.

Prefers the cells are at least during steps a), b) and / or d), at least one added to the cell culture medium Cell migration potentially affecting additive or a physical influence, in particular a radiation or a Temperature change, exposed. The cells can also, in particular before step d), preferably before the step c), in particular before step b), particularly preferably before the step a), be subjected to a modification. In the modification can it is a genetic, pharmacological, physical, biochemical or chemical modification. As cells become preferred in the method according to the invention Used fibroblasts, endothelial cells or epithelial cells. at These and other cells can be stable cells act transfected cell line.

following The invention is based on an embodiment and the figures explained in more detail. Show it:

1 a photographic representation of a device according to the invention,

2a C is a side view of a barrier, a view of the underside of the barrier and a view of the top of the device according to the invention,

3 microscopic images of a study of the migration of human tubular epithelial cells,

4 a quantitative representation of the migration of human tubulus ephithelium cells after 24 hours,

5 a schematic representation for explaining the method according to the invention for investigating a migration of adherent cells,

6a -C a comparison of the coating with matrix proteins in the scratch assay and the inventive method and

7 Schematic drawing of a fibronectin-coated support and the position of a cell culture insert from ibidi GmbH on this support and microscopic images of the fibronectin coating after removal of the cell culture insert at various positions marked in the schematic drawing.

1 is a photographic representation of the device according to the invention 10 , The bottom of a plastic Petri dish serves 11 as a carrier 25 which is coated with a protein. Furthermore, photography shows the barrier 12 . those with the bottom of the petri dish 11 in contact. The barrier 12 has two oppositely disposed from bent wire springs 14 on. The feathers 14 allow a clamping of the barrier 12 at the edge of the Petri dish 11 so that the barrier 12 to the bottom of the Petri dish 11 is pressed.

2a shows the barrier 12 with springs made of plastic or metal 14 in a side view. 2 B shows the barrier 12 in a supervision on the for the contact with the carrier 25 provided contact surface 16 , 2c shows the device according to the invention 10 in top view. The bottom of a Petri dish forms 11 the carrier 25 , The barrier 12 is by means of the springs 14 on the surrounding wall 15 the Petri dish 11 so clamped that the barrier 12 to the bottom of the Petri dish 11 is pressed.

The process according to the invention is carried out as follows:
The barrier 12 is first sterilized in 70% ethanol and then completely dried. It then becomes by means of the springs 14 in the Petri dish 11 , which has a diameter of 35 mm and whose bottom is coated with fibronectin, fixed so that it is pressed onto this soil. Subsequently, 600,000 cells of the human renal tubular epithelial cell line HKC-8 in 2 ml cell culture medium in the Petri dish 11 seeded and cultured in an incubator for 14 hours. After this time, a confluent monolayered cell lawn has developed 22 around the barrier 12 formed around.

The experiment can also be performed with any other adherently growing cell line. The number of cells to be sown and the time required to create a confluent cell lawn 22 are to be determined empirically for each cell type.

In the experiment carried out here, the barrier is at the end of the mentioned 14 hours 12 from the Petri dish 11 away. The geometry of the contact surface 16 sets a defined cell-free area 18 with a cell-free strip 24 firmly. To determine the migration velocity v, different regions are formed over the entire length of the cell-free strip 24 used. On the back of the carrier 25 , here the Petri dish 11 , can, for. B. by scoring with a razor blade, marks 20 be attached. These marks 20 allow orientation to the same areas of the strip at different times 24 capture and analyze. For this purpose, by means of a photomicroscope at different times immediately above and below the marked locations two shots of the cell-free strip 24 made.

3 shows recordings of said human tubulus epithelial cells at time 0 (left) and 24 hours later (right). The upper two images show a control experiment in which the cell culture medium contained no additive. In the two lower illustrations, the cell culture medium contained in each case the Rho-kinase inhibitor Y-27632 in a concentration of 10 μM. The comparison of the images shows that Y-27632 causes a significantly increased migration speed. The results of this experiment are in 4 shown graphically. The height of the bars indicates the average migration speed v in microns per hour.

5 Figure 12 shows a schematic representation of the analysis of cell migration in the cell-free strips 24 into it. In the Petri dish on the top left 11 is one in the confluent cell lawn 22 through the barrier 12 kept cell-free area 18 with the cell-free strip 24 shown. On the back of the Petri dish 11 are two markings 20 appropriate. You can also attach more than two markers. The markings are used for orientation when selecting the image sections 21 which are taken photographically at the beginning and at one or more later times. At the beginning and at a later date, here 24 hours after removing the barrier 12 , is the average width b of the cell-free strip 24 calculated by using a computer program, the cell-free area A and the height h is determined. The average width b of the area A results as a quotient of the cell-free area A and the height h. The difference Δb between the average width b at the start of migration, ie immediately after removal of the barrier 12 , and the average width b after the time t, here 24 hours, is used to calculate the migration velocity v. For this purpose, the difference Δb is divided by twice the time t. The factor 2 takes into account the fact that cells from two sides in the cell-free strip 24 immigrate and thereby reduce the cell-free area A. The formula for the migration speed v is as follows: v = Δb / 2t

The Analysis of the migration can be done after given periods of time t or done by real-time video microscopy. Real-time video microscopy allowed also the analysis of the migration of single cells starting from one Cell cluster.

6a to 6c show immunofluorescence microscopy images of each one with the extracellular matrix protein fibronectin coated carrier 25 after immunostaining of fibronectin. At the in 6a presented trial HKC-8 cells were on the coated support 25 seeded. After reaching confluency, a strip was scratched out. In the in the 6b and 6c HKC-8 cells were placed on the coated support 25 , the component of a device according to the invention 10 with a barrier 12 was seeded. Upon reaching confluence, the barrier became 12 away. With the carriers 25 from both experiments, an immunostaining specific for fibronectin was performed. 6b shows the cell-free strip 24 ("Gap") under the contact surface 16 the barrier 12 , 6c shows the cell-free strip 24 with adjacent cells. A comparison of 6a with the 6b and 6c shows that the fibronectin is scraped off in the scratch assay. The cells thereby migrate during the scratch assay into a cell-free region which has no coating, while in the process according to the invention they migrate into the cell-free region on the envisaged coating, here fibronectin.

7 shows immunofluorescence micrographs of a fibronectin-coated carrier 25 after an immunostaining specific for fibronectin. On the carrier 25 became the cell culture use 26 set up by ibidi GmbH. The cell culture use 26 on the carrier 25 is shown schematically in the drawing shown below left. The rectangles marked with 1 to 5 show areas in which the immunofluorescence microscopy images labeled with the same numeral after removal of the cell culture insert 26 have been made. It shows clearly that the removal of the cell culture use 26 The company ibidi GmbH leads to a replacement of the fibronectin coating and thus in the study of a migration of adherent cells to an artificial result.

10

contraption

11

Petri dish

12

barrier

14

feather

15

wall the Petri dish

16

contact area

18

cell-free area

20

mark

21

image section

22

cell lawn

24

cell-free strip

25

carrier

26

Cell culture insert

A

area

b

average width

H

height

t

Period of time

v

migration rate

QUOTES INCLUDE IN THE DESCRIPTION

This list The documents listed by the applicant have been automated generated and is solely for better information recorded by the reader. The list is not part of the German Patent or utility model application. The DPMA takes over no liability for any errors or omissions.

Cited non-patent literature

• Todaro GJ et al., J. Cell. Physiol. 1965, 66 (3): 325-33 [0002]

Claims (25)

Contraption ( 10 for investigating a migration of adherent cells on a coating of a carrier consisting of at least one protein ( 25 ), at least one with the carrier ( 25 ) and by the wearer ( 25 ) removable barrier ( 12 ) provided on the support ( 25 ) prevents growth of the cells in the region of the contact, so that when the cells are cultivated on the support ( 25 ) remains an area kept clear of the cells, whereby removal of the barrier ( 12 ) allows the cells to migrate into the reserved area, the barrier ( 12 ) on the one with the carrier ( 25 ) is made of a material which does not adhere to the coated support ( 25 ) adheres. Contraption ( 10 ) according to claim 1, wherein the carrier ( 25 ) is the bottom of a cell culture vessel or placed on the bottom of a cell culture vessel. Contraption ( 10 ) according to any one of the preceding claims, wherein the carrier ( 25 ) is a plate, in particular of glass or a first plastic. Contraption ( 10 ) according to claim 2 or 3, wherein the cell culture vessel comprises a, in particular from a second plastic, Petri dish ( 11 ) or a well of a microtiter plate. Contraption ( 10 ) according to one of the preceding claims, wherein the barrier ( 12 ) consists of a, in particular solid, biocompatible material, preferably a third plastic and / or a, preferably inert, metal, in particular stainless steel. Contraption ( 10 ) according to one of the preceding claims, wherein the third plastic, in particular ultra-high molecular weight, polyethylene (PE), polystyrene (PS), polypropylene (PP), Teflon, polymethyl methacrylate (PMMA), acrylonitrile-butadiene-styrene copolymer (ABS), polyethylene terephthalate ( PET), polyphenylene ether (PPE), styrene / acrylonitrile (SAN), styrene / butadiene (SB) or another plastic used in medicine or biomedicine. Contraption ( 10 ) according to one of the preceding claims, wherein, in particular at the barrier ( 12 ), the carrier ( 25 ) or the cell culture vessel, at least one means is provided, by means of which the barrier ( 12 ) in the area of contact with the wearer ( 25 ) is pressed. Contraption ( 10 ) according to any one of the preceding claims, wherein the means comprises one at the barrier ( 12 ), the carrier ( 25 ) or the cell culture vessel, in particular formed from the or a further metal or the third or a fourth plastic, spring ( 14 ) or clip for clamping the barrier ( 12 ) on the carrier ( 25 ) or the cell culture vessel, or is a magnet. Contraption ( 10 ) according to claim 8, wherein the spring ( 14 ) and the barrier ( 12 ) or the clasp and the barrier ( 12 ) are integrally formed. Contraption ( 10 ) according to one of the preceding claims, wherein the barrier ( 12 ) is produced in an injection molding process. Contraption ( 10 ) according to one of the preceding claims, wherein the barrier ( 12 ), in particular on the one with the carrier ( 25 ), or has no coating of the or another protein or is such that it does not assume such a coating. Device according to one of the preceding claims, wherein the surface of the barrier ( 12 ), in particular the surface on which with the carrier ( 25 ), preferably by physical or chemical modification of the surface or by application of a preferably inert coating. Contraption ( 10 ) according to any one of the preceding claims, wherein the carrier ( 25 ) and the barrier ( 12 ) or the cell culture vessel and the barrier ( 12 ) in the region of the contact have in their form corresponding, in particular planar, surfaces. Contraption ( 10 ) according to one of the preceding claims, wherein the barrier ( 12 ) is shaped so that the area kept clear of the cells, in particular 0.1 to 3 mm, preferably 0.5 to 2 mm wide strip or one, in particular 0.1 to 3 mm, preferably 0.5 to 2 mm diameter, circle is. Contraption ( 10 ) according to claim 14, wherein the barrier ( 12 ) is shaped so that the area kept clear of the cells is widened at both ends of the strip. Contraption ( 10 ) according to any one of the preceding claims, wherein the protein is an extracellular matrix protein, in particular a fibronectin or a collagen. Method for investigating migration of adherent cells by means of a device ( 10 ) according to one of the preceding claims, comprising the following steps: a) sowing the cells on the support ( 25 ) in one Cell culture medium, b) culturing the cells until they are in a barrier ( 12 ) are confluent, c) removing the barrier ( 12 and detecting the extent of the area vacated by the cells, and d) further culturing the cells and detecting a change in the extent of the area kept clear of the cells. The method of claim 17, wherein detecting the Extent of the area kept free of the cells and of its change by means of, in particular, regular, time intervals performed photographic Recordings or by means of, in particular in real time, video microscopy he follows. The method of claim 17 or 18, wherein an analysis the migration of the cells takes place. The method of claim 19, wherein the analysis is a Analysis of a migration speed v, a migration direction, a protein expression, a release of a messenger substance and / or protein localization. The method of claim 20, wherein the analysis of Protein expression and / or protein localization immunocytochemical he follows. Method according to one of claims 17 to 21, wherein the cells, at least during steps a), b) and / or d), at least one added to the cell culture medium Cell migration potentially affecting additive or a physical influence, in particular a radiation or a temperature change, get abandoned. Method according to one of claims 17 to 22, wherein the cells, in particular before step d), preferably before the step c), in particular before the step b), particularly preferred before step a), be subjected to a modification. The method of claim 23, wherein the modification a genetic, pharmacological, physical, biochemical or chemical modification. Method according to one of claims 17 to 24, wherein the cells are fibroblasts, endothelial cells or epithelial cells are.