CN110205243A - The manufacturing method of cell attachment device and the cell attachment device - Google Patents

Abstract

The present invention provides a kind of cell attachment device, lies in a surface of a substrate, forms a cell attaching layer, and being formed then at the surface of the cell attaching layer has the magnetic warming layer of a plurality of warmings high molecular one, and each warming macromolecule has a plurality of magnetic particles.When variation effect of the magnetism warming layer in a physical features, so that a plurality of cells are able to be attached on the cell attaching layer or go to attach by the cell attaching layer.Cell attachment device through the invention can formerly control temperature to one first temperature and lure that the warming macromolecule on substrate generates structure change into, make cell attachment on the cell attaching layer.The separately magnetic particle temperature rise with alternating magnetic field (HAC) to making in warming macromolecule enables cell to maintain the effect of cell attachment in the second temperature lower than first temperature.

Description

Cell-attached device and method for manufacturing the cell-attached device

technical field

The present invention relates to a cell attachment technology, in particular to a cell attachment device and a cell attachment device which utilizes physical characteristic changes in an alternating magnetic field to change the temperature-sensitive polymer with magnetic particles to produce structural changes so that cells can be attached to a substrate method and its method of manufacture.

Background technique

In normal tissues, homotypic cells and heterotypic cells must grow in corresponding positions in order to have the functionality of normal tissues. Therefore, cell patterning technology is an important research topic in the field of biomedical engineering. The concept of patterned cell arrangement was proposed in the 1960s; basically, patterned cell arrangement is divided into two types: passive and active.

The passive method mainly relies on the combination of cell-adhesion ligands arranged on the substrate and receptors on the cell surface to achieve cell arrangement. This method takes a long time to respond and cannot be reused. The active method mainly relies on applying an external force such as sound waves, optical clamps, and dielectrophoresis to guide cells to defined positions for patterned arrangement. Although these active cell arrangement methods are reversible, have good reaction efficiency and short operation time, they still have some limitations in application. For example, the microporous structure is easy to cause changes in the intracellular skeleton of cells during the attachment process. , and cannot screen the target cells for graphical arrangement. Although dielectrophoresis technology can be used to manipulate population cells, it must be operated in a low-conductivity culture environment, and high-frequency currents are required to manipulate cells to avoid damage to cell membranes. In addition, the preparation of a large number of microelectrodes has also increased. Although the optical clamp can be used to manipulate a single cell, it is limited by the relatively expensive instrument and the limitation of operating power to prevent thermal damage to the cell.

In the prior art, for example, Tang et al. published "Temperature-Responsive Polymer Modified Surface for Cell Sheet Engineering" in 2012, Polymer, 15Aug, 2012, 4, 1478-1498, which proposed a temperature-responsive polymer Techniques for cell attachment. On the surface of the cell attachment layer (tissue culture polystyrene, TCPS), a temperature-sensitive polymer layer that can change the hydrophilicity and hydrophobicity by changing the temperature is formed, which is composed of poly N-isopropylacrylamide (poly(N-isopropylacrylamide), poly (NIPAAm)). However, although the temperature-sensitive polymer layer can form a hydrophobic state when it is higher than the lower critical solution temperature (lower critical solution temperature, LCST), so that cells can be attached to TCPS, on the contrary, when the temperature is lower than LCST, The temperature-sensing polymer layer turns into a hydrophilic state, which makes the cells on the TCPS deattach.

Although in the prior art, the cell attachment device that can produce the effect of attaching cells, the phenomenon of cell attachment must be maintained at a temperature higher than that of the LCST, and the cell attachment effect cannot be maintained at normal temperature. It is difficult to arrange these cells into specific patterns. Although there are cell arrangement technologies in the aforementioned prior art, they all have their deficiencies. Based on the foregoing, a cell attachment device and method are needed to solve the problems of the prior art.

Contents of the invention

The object of the present invention is to provide a cell-attached device and a method for manufacturing the cell-attached device, so as to solve the above-mentioned technical problems existing in the prior art.

In order to achieve the above object, the technical scheme adopted in the present invention is:

A cell attachment device is characterized in that it includes:

a substrate;

a cell attachment layer formed on the surface of the substrate; and,

A magnetic temperature-sensing layer formed on the surface of the cell attachment layer, the magnetic temperature-sensing layer has a plurality of magnetic temperature-sensing polymers, each magnetic temperature-sensing polymer has a temperature-sensing polymer and a plurality of magnetic particles, when When a physical characteristic of the magnetic temperature-sensing layer changes, a plurality of cells can be attached to the cell-attached layer or deattached from the cell-attached layer.

The cell attachment device, wherein: the cell attachment layer is polyethyleneimine, and the temperature-sensitive polymer is poly(N-Isopropylacrylamide).

Said cell attachment device, wherein: the object is characterized by temperature, magnetic field or a combination of both.

The cell-attached device further has a magnetic structure, the magnetic structure has a pattern structure, and the magnetic structure is arranged between the cell-attached layer and the substrate or on a side outside the substrate.

Said cell attachment device, wherein: the pattern structure is a geometric pattern structure with a plurality of concentric arrangements.

A method for manufacturing a cell attachment device, characterized by comprising the following steps:

Manufacturing a plurality of magnetic temperature-sensitive polymers, each magnetic temperature-sensitive polymer has a first temperature-sensitive polymer, and each first temperature-sensitive polymer has a plurality of magnetic particles;

providing a substrate, and forming a cell adsorption layer on the substrate; and,

Using the plurality of magnetic temperature-sensitive polymers to form a magnetic temperature-sensitive layer on the cell adsorption layer, when a physical characteristic applied to the magnetic temperature-sensitive layer changes, a plurality of cells can be attached to the cell-attached layer Or deattach by the cell adsorption layer.

The method for manufacturing the cell attachment device, wherein: before forming the cell adsorption layer, it also includes forming a magnetic thin film layer on the substrate, which has a pattern structure, and when forming the magnetic temperature-sensing layer, the A plurality of magnetic thermosensitive polymers are arranged corresponding to specific positions on the pattern structure.

The method for manufacturing the cell-attached device, wherein: it also includes a permanent magnet with a specific pattern on one side of the substrate, which has a pattern structure, and when forming the magnetic temperature-sensing layer, the plurality of permanent magnets with The temperature-sensitive polymers of the magnetic particles are arranged along the outline of the pattern structure.

The method for manufacturing the cell attachment device, wherein: the object is characterized by a combination of temperature and magnetic field, and the plurality of magnetic temperature-sensitive polymers are in a contracted state when the cell attachment device is at a first temperature, so that the cells are adsorbed on On the cell adsorption layer, at a second temperature, by virtue of the change of the magnetic field, only the temperature-sensitive polymer corresponding to the pattern structure maintains a contracted state, so that cells can be placed on a specific area corresponding to the pattern structure Adsorbed by the cell adsorption layer.

The manufacturing method of the cell attachment device, which further includes forming a plurality of second temperature-sensitive polymers, and when forming the magnetic temperature-sensitive layer, will contain the magnetic temperature-sensitive polymers and the second temperature-sensitive The polymer mixed solution is coated on the cell adsorption layer, so that on the area corresponding to the pattern structure, the magnetic temperature-sensitive polymer is formed on the cell adsorption layer, and on other areas, it has the second high temperature sensitivity. Molecules are formed on the cell-adsorbed layer.

Compared with prior art, the beneficial effect that the present invention has is:

The present invention provides a cell attachment device and method, which utilizes the temperature-sensing polymer to have magnetic particles to form a magnetic temperature-sensing polymer. The magnetic temperature-sensing polymer can attach cells through temperature changes, and can further The control of the variable magnetic field enables the cells to maintain the attached state under the condition of lower than the LCST temperature, so as to avoid the problem of damage to the cells caused by the attachment of the cells by external force in the prior art, as well as the occurrence of problems that are not used at low temperatures. The effect of allowing cells to attach or deattach under LCST conditions.

The present invention provides a cell attachment device and method. In addition to allowing cells to attach or deattach under the condition of controlling the temperature change of LCST, it can also use an external magnetic field to control the arrangement pattern of cells, thereby simulating various Different tissue cells to increase the field of its application. In particular, through the structure of the magnetic film layer with a specific pattern, the temperature-sensitive polymer with magnetic particles can be attached to the specific position of the pattern structure, and then the adhesion of the attached cells can be controlled by the alternating magnetic field. location and area, forming a specific cell pattern.

The invention provides a cell attachment device and method, through the structure of a magnetic temperature-sensitive layer or a magnetic temperature-sensitive layer and a magnetic film layer, the attached cells can be attached without injecting magnetic particles. The effect of de-attachment can maintain the integrity of cells and expand the application range of research and development.

Description of drawings

Fig. 1 is a schematic cross-sectional view of the first embodiment of the cell attachment device of the present invention.

2A and 2B are schematic diagrams showing changes in hydrophilicity and hydrophobicity of temperature-sensitive polymers at temperatures above and below the LCST.

FIG. 2C and FIG. 2D are schematic diagrams showing changes in the hydrophilicity and hydrophobicity of the temperature-sensitive polymer with magnetic particles under the action of an alternating magnetic field when the temperature is lower than the LCST temperature.

3A to 3D are schematic flow charts of manufacturing the cell attachment device of the present invention.

Fig. 4A is a schematic cross-sectional view of the second embodiment of the cell attachment device of the present invention.

FIG. 4B is a schematic diagram of the pattern structure of the magnetic thin film.

5A and 5B are schematic diagrams of the cell attachment device of the present invention for attaching cells and forming specific cell patterns.

Fig. 6 is a schematic diagram of another embodiment of the magnetic attachment device of the present invention.

As shown in FIG. 7A to FIG. 7F , the figure is a schematic flowchart of the second embodiment of the cell attachment device in FIG. 4A and FIG. 4B .

Description of reference signs: 2, 2a-cell attachment device; 20-substrate; 21-cell attachment layer; 22-magnetic temperature-sensing layer; 23-magnetic temperature-sensing polymer; 230-temperature-sensing polymer; 231-magnetic Particles; 25-magnetic film layer; 250-pattern structure; 26-temperature-sensitive layer; 27-permanent magnet; 3, 3a-cell; 90-alternating magnetic field.

Detailed ways

Various exemplary embodiments will be described more fully hereinafter with reference to the accompanying drawings, in which some exemplary embodiments are shown. However, inventive concepts may be embodied in many different forms and should not be construed as limited to the illustrative embodiments set forth herein. Rather, these exemplary embodiments are provided so that this disclosure will be thorough and complete, and will fully convey the scope of the inventive concept to those skilled in the art. Like numbers indicate like elements throughout. The cell-attaching device and method and the manufacturing method of the cell-attaching device will be described below with various embodiments and drawings, however, the following embodiments are not intended to limit the present invention.

Please refer to FIG. 1 , which is a schematic cross-sectional view of the first embodiment of the cell attachment device of the present invention. In this embodiment, the cell attachment device 2 includes a substrate 20 , a cell attachment layer 21 and a magnetic temperature sensing layer 22 . The material of the substrate 20 is not limited. A glass substrate, a silicon substrate or a flexible substrate such as a plastic substrate can be used, but it is not limited thereto. A cell attachment layer 21 is formed on the upper surface of the substrate 20 for providing cells to attach thereto. In one embodiment, the cell attachment layer 21 may be polyethyleneimine (polyethyleneimine, PEI), but not limited thereto, and those skilled in the art can select an appropriate cell attachment layer material according to requirements.

The upper surface of the cell attachment layer 21 is formed with the magnetic temperature sensing layer 22, which can act on the change of the characteristics of the external object of the cell attachment device 2, such as: changing the ambient temperature or providing an alternating magnetic field ( HAC), changing its hydrophilicity and hydrophobicity. The magnetic temperature-sensing layer 22 has a plurality of magnetic temperature-sensing polymers 23 , each magnetic temperature-sensing polymer 23 has a magnetic temperature-sensing polymer 230 , and a plurality of magnetic particles 231 are attached on its surface or inside it. In one embodiment, the magnetic thermosensitive polymer 23 is poly N-isopropylacrylamide (poly(N-Isopropylacrylamide), poly(NIPAAm)), but it is not limited thereto. The magnetic particles 231 are nano magnetic particles in one embodiment. 2A and 2B, in this embodiment, the magnetic temperature-sensing layer 22 has a low critical solution temperature (lower critical solution temperature, LCST), in one embodiment, it is between 32-34.4 ° C , the present embodiment is 32° C. It should be noted that the LCST temperature is determined according to the properties of the material, and is not limited by the present embodiment.

In FIG. 2A, when the ambient temperature is higher than the LCST, for example: 37°C, each magnetic temperature-sensitive polymer 23 in the magnetic temperature-sensitive layer 22 will become hydrophobic and shrink, so that the originally expanded magnetic temperature-sensitive The cell attachment layer 21 covered by the sensitive polymer 23 will be exposed, as shown in area A in FIG. 2A . At this time, if there are cells 3 in the environment, in one embodiment, the environment is liquid, and the cells 3 will be attached to the cell attachment layer 21 . Conversely, as shown in Figure 2B, when the ambient temperature is lower than the LCST, for example: 28°C, the magnetic temperature-sensing polymer 23 of the magnetic temperature-sensing layer 22 will change from hydrophobicity to hydrophilicity, and its structure will expand to cover the original In area A, the cells 3 are deattached and detached from the cell attachment layer 21 .

As shown in FIGS. 2C and 2D , they are schematic diagrams of changing the hydrophilicity and hydrophobicity of the magnetic temperature-sensing layer through the alternating magnetic field 90 . In FIG. 2C , when the ambient temperature is lower than the LCST temperature, for example, at room temperature of 28°C, although it is logical that the magnetic temperature-sensitive polymer 23 will become hydrophilic and expand, thereby covering the cell attachment layer 21, but The alternating magnetic field 90 acts on the magnetic particles 231 of the magnetic temperature-sensitive polymer 23, as shown in FIG. The temperature of the magnetic temperature-sensing polymer 23 is heated up, so that the magnetic temperature-sensing polymer 23 with the magnetic particles 231 produces a hydrophobic state, that is, the structure of the temperature-sensing polymer shrinks, and then the cells 3 are attached to the cell attachment layer 21 or Keep the cells 3 attached to the cell attachment layer 21 continuously. By virtue of the control of the alternating magnetic field 90, it is possible to maintain or change the state of the cells 3 attached to the cell attachment layer 21 from the situation where the cells 3 should be deattached at room temperature, greatly increasing the number of cell attachment devices. 2 application environment. For example, in a use situation, the user does not need to deliberately maintain the temperature of the environment to maintain the cells attached to the cell attachment layer 21. Even at normal temperature, the effect of the alternating magnetic field can maintain the temperature. The effect of the cells being attached can reduce the cost of maintaining the ambient temperature, including equipment and energy, and can also increase the convenience of transportation.

Next, the method of fabricating the structure shown in FIG. 1 will be described. Please refer to FIG. 3A and FIG. 3D , which are schematic flow charts for fabricating the cell attachment device of the present invention. First, perform step 30 to convert the temperature-sensitive polymer material into an aminated temperature-sensitive polymer material. In this step, dissolve the temperature-sensitive polymer material, such as poly(NIPAAm), and hydrazine N2H2 in methanol, and heat to Reflux at 90°C, cool down to room temperature after the reaction, freeze-dry after dialysis to obtain aminated poly(NIPAAm). Next, go to step 31 to generate the magnetic temperature-sensitive water-gel material. In this step, carbodiimide (1-Ethyl-3-(3-dimethylaminopropyl)-carbodiimide, EDC) and N-hydroxysuccinimide (N-hydroxysuccinimide, NHS) were mixed with 5, 10, 20 Dissolve 40wt% magnetic nanoparticles CM-dextran/Fe3O4 in deionized water, apply EDC/NHS to activate the COOH functional group of CM-dextran, and then add the aminated poly(NIPAAm) obtained in step 31 to make the two or grafted to obtain a magnetic temperature-sensitive water-gel material, a schematic diagram of the structure of a magnetic temperature-sensitive polymer 23 with magnetic particles formed by it, as shown in Figure 3A, the structure of the polymer 23 is nearly spherical, and the magnetic nanoparticles 231 Grafted on the surface of magnetic thermosensitive polymer 23.

As shown in FIG. 3B , proceed to step 32 to provide a substrate 20 whose features are as described above and will not be repeated here. Next, proceed to step 33, the step of forming a cell attachment layer. As shown in FIG. 3C , in step 33 , the substrate is soaked in 1.0 wt % PEI (polyethyleneimine) solution, and dried, for example, at room temperature, to form a cell attachment layer with a film thickness of about 1 μm. Next, step 34 is performed to form the magnetic temperature-sensing polymer 23 on the cell attachment layer to form the magnetic temperature-sensing layer 22 . In this step, the deionized water containing the structure in Figure 3A is dropped on the substrate in Figure 3B, and the negatively charged poly (NIPAAm) and the positively charged PEI are closely attracted by the positive and negative forces, and the spin coating is continued. The poly(NIPAAm) is evenly coated on the substrate 20 to complete the poly(NIPAAm) magnetic temperature-sensing layer 22 , as shown in FIG. 3D .

Please refer to FIG. 4A , which is a schematic cross-sectional view of the second embodiment of the cell attachment device of the present invention. In this embodiment, the cell-attached device 2a is basically similar to that of FIG. 1, the difference is that the cell-attached device 2a of this embodiment has a magnetic The film layer 25 has a pattern structure 250 so that the plurality of magnetic thermosensitive polymers 23 with magnetic particles 231 are arranged along the outline of the pattern structure 250 . In one embodiment, as shown in FIG. 4B , the pattern structure 250 is composed of a plurality of concentric geometric structures, which are rectangular structures in one embodiment. It should be noted that the structure of the geometric pattern is determined according to the needs of users, and is not limited to the concentric geometric pattern of this embodiment. In addition, in one embodiment, on the cell attachment layer 21 in the non-corresponding geometric pattern area 251 , a temperature-sensing layer 26 composed of a magnetic temperature-sensing polymer 23 without magnetic particles 231 is formed.

Through the structure shown in FIG. 4A, the pattern formed after the cell attachment device 2a is attached can be controlled, so that the cell attachment device 2a can simulate cells with specific functions, such as liver, heart, etc., and the application field can be increased. Contribute to the development of drugs or treatments. Taking the structure shown in FIG. 4A and FIG. 4B as an example, under conditions higher than the LCST temperature, for example: 37°C, as in the state shown in FIG. 2A above, since the magnetic temperature-sensitive polymer 23 is hydrophobic, the substrate at this time The structure of the magnetic temperature-sensitive polymer 23 shrinks, so that the cells 3 can be attached to the cell attachment layer 21, forming a state as shown in FIG. 5A . When the ambient temperature is lower than the LCST of the substrate, for example: 27° C., as shown in FIG. 2B , the structure of the magnetic thermosensitive polymer 23 expands to induce the cells 3 to detach from the cell attachment layer 21 . At this time, by applying an alternating magnetic field (HAC), the temperature of the magnetic temperature-sensitive polymer 23 with the magnetic particles 231 increases because the magnetic particles 231 are subjected to the action of the alternating magnetic field (HAC), and then the temperature of the magnetic temperature-sensitive polymer 23 with the magnetic particles 231 can be increased. The magnetic temperature-sensitive polymer 23 maintains its hydrophobic state, so that the cells 3 attached to it continue to maintain the attached state. As for other magnetic temperature-sensitive polymers 23 that do not correspond to the pattern structure of the magnetic thin film layer 25, since the temperature has been lower than the LCST, the structure of the magnetic temperature-sensitive polymer 23 has changed and become hydrophilic, so that it was originally attached to The cells 3 a of the cell-attached layer 21 are detached from the cell-attached layer 21 . With the above-mentioned environmental temperature conditions higher than the LCST temperature, the cells are all attached to the cell attachment layer 21, and then by controlling the environmental temperature conditions to be lower than the LCST, and then applying an alternating magnetic field, the cells in a specific area 3 maintained on the cell-attached layer 21, thereby producing a cell-attached device 2a with a specific pattern of attached cells.

Next, the principle of patterned cell arrangement will be described. Since in this embodiment, the pattern structure is a plurality of concentric rectangular structures, therefore, a magnetic temperature-sensitive polymer 23 with a plurality of magnetic particles 231 is formed along the outline of the rectangular structure. . When the temperature is lower than the LCST and the alternating magnetic field HAC is applied, the cells are attracted by the dissipation field generated at the turning point of the concentric rectangular structure, so that the cells 3 will be concentrated on the corresponding temperature sensor with magnetic particles at the turning point of the pattern structure 250 On the polymer, a patterned cell culture plate is formed as shown in Figure 5B. In addition, when the cell attachment device is placed at room temperature without HAC exposure to the alternating magnetic field, all cells can be deattached to achieve another application.

Please refer to FIG. 6 , which is a schematic diagram of another embodiment of the magnetic attachment device of the present invention. In this embodiment, it is basically similar to FIG. 4A , the difference is that this embodiment uses a permanent magnet 27 with a specific shape or pattern to replace the magnetic film layer 25 in FIG. 4A . In the same way, the magnetic temperature-sensitive polymer 23 with magnetic particles 231 is formed at the position corresponding to the permanent magnet 27 , and the magnetic temperature-sensitive polymer 23 without magnetic particles is formed in the rest of the area. As for the way of using the alternating magnetic field to control the cell attachment, it is as mentioned above, so it will not be repeated here.

Next, the method for attaching cells of the present invention is described. Firstly, the cell attachment device 2a or 2b as shown in FIG. 4A or FIG. 6 is provided in step 40 . Then proceed to step 41, at a first working temperature, a plurality of cells are attached to the cell attachment device 2a. In one embodiment, the working temperature is higher than the LCST temperature of the temperature-sensitive polymer, which is 37° C. in this embodiment, but it is not limited thereto. Under the first working temperature, the cells attached to the cell attachment device 2a result in the state shown in FIG. 5A , that is, the cells 3 are covered on the surface of the cell attachment device with the temperature-sensitive polymer. Then proceed to step 42, providing an alternating magnetic field HAC to act on the cell attachment device 2a. Next, in step 43, the temperature is lowered from the first working temperature to a second working temperature, and through the action of the alternating magnetic field, at least a part of the plurality of cells is attached to the cell attachment device 2a, To form a patterned cell attachment structure as shown in Figure 5B. In one embodiment, the second working temperature is lower than the LCST temperature of the temperature-sensitive polymer, 28° C. in this embodiment.

Please refer to FIGS. 7A-7F , which are schematic flow charts showing the structure of the second embodiment of the cell attachment device in FIGS. 4A and 4B . In this embodiment, step 50 is first performed to fabricate the temperature-sensitive material poly(NIPAAm). In one embodiment, NIPAAm and chain transfer agent (methyl3-mercaptopropionate) are added into a reaction flask containing 40 mL of deionized water, nitrogen gas is blown in, and heating is performed. The initiator potassium persulfate (potassium peroxydisulfate, KPS) was added to react and polymerize to obtain the temperature-sensitive material poly(NIPAAm). Then proceed to step 51 , making a magnetic temperature-sensitive water-gel material composed of temperature-sensitive polymers containing magnetic particles. In one embodiment, the temperature-sensitive polymer material and hydrazine N2H2 in step 50 are dissolved in methanol, heated to 90°C to generate reflux, cooled to room temperature after the reaction, and freeze-dried after dialysis to obtain the aminated poly( NIPAAm). Then, carbodiimide (1-Ethyl-3-(3-dimethylaminopropyl)-carbodiimide, EDC) and N-hydroxysuccinimide (N-hydroxysuccinimide, NHS) were mixed with 5, 10, 20 and 40wt % magnetic nanoparticles CM-dextran/Fe3O4 are dissolved in deionized water, and the COOH functional group of CM-dextran is activated by EDC/NHS, and then aminated poly(NIPAAm) is added to graft the two to obtain a magnetic temperature sensor Type water gel material, the polymer 23 has a nearly spherical structure, and magnetic nanoparticles 231 are grafted on the surface of the magnetic temperature-sensitive polymer 23 .

Then proceed to step 52, coating the AZ photoresist 200 on the substrate 20 to form the structure shown in FIG. 7A, and then cover the photomask 201 for UV photoetching as shown in FIG. , forming the structure shown in Figure 7C. As shown in Figure 7D, the photoresist 200 is removed, and the patterned magnetic thin film is fixed on the glass substrate. The pattern is a concentric square structure, and the line width and spacing are both 12 μm. The substrate can be magnetized with an external magnetic field to generate a magnetic dissipation field. . Continue to step 53, soak the glass substrate on which the magnetic thin film is fixed in 1.0 wt% PEI (polyethyleneimine) solution, and dry naturally at room temperature to form a cell attachment layer 21 with a thickness of about 1 μm, as shown in FIG. 7E Show. Continue to step 54, as shown in FIG. 7F , and then drop the mixed solution of the temperature-sensitive water-colloid solution and the magnetic temperature-sensitive water-colloid solution onto the cell attachment layer 21 of the substrate 20 for spin coating. During the drying process of the solution, the magnetic temperature-sensitive polymer 23 containing magnetic particles 231 will be attracted to the patterned structure of the magnetic film layer 21, while the magnetic temperature-sensitive polymer 23 without magnetic particles 231 will be attached to cells without a patterned structure After waiting for drying on the attachment layer 21, the preparation of the cell attachment device as shown in FIG. 4A is completed.

The above description is only illustrative of the present invention, rather than restrictive. Those of ordinary skill in the art understand that many modifications, changes or equivalents can be made without departing from the spirit and scope defined in the claims, but All will fall within the protection scope of the present invention.

Claims (10)

1. a kind of cell attachment device, it is characterized in that including:

One substrate；

One cell attaching layer is formed in the surface of the substrate；And

One magnetic warming layer, is formed in the surface of the cell attaching layer, which has a plurality of magnetic warming high scores Son, each magnetism warming macromolecule has a warming macromolecule and a plurality of magnetic particles, when acting on the magnetism warming layer A physical features variation when, a plurality of cells are able to be attached on the cell attaching layer or go to attach by the cell adsorption layer.

2. cell attachment device according to claim 1, it is characterised in that: the cell attaching layer is polyethyleneimine (polyethylenimine), which is poly-N-isopropyl acrylamide (poly (N- Isopropylacrylamide))。

3. cell attachment device according to claim 1, it is characterised in that: the object features are temperature, magnetic field or both Combination.

4. cell attachment device according to claim 1, it is characterised in that: also have a magnetic texure, the magnetic texure With a patterning, which is set between the cell attaching layer and the substrate or in being set to outside the substrate Side.

5. cell attachment device according to claim 4, it is characterised in that: the patterning is with a plurality of concentric rows The geometrical pattern structure of column.

6. a kind of manufacturing method of cell attachment device, it is characterized in that including the following steps:

A plurality of magnetic warming macromolecules are manufactured, each magnetic warming macromolecule has an one first warming macromolecule, and every 1 the One warming macromolecule has a plurality of magnetic particles；

One substrate is provided, and forms a cell adsorption layer on the substrate；And

Using a plurality of magnetic warming macromolecules in forming a magnetic warming layer on the cell adsorption layer, when acting on the magnetism When the physical features variation of warming layer, a plurality of cells are able to be attached on the cell attaching layer or are gone by the cell adsorption layer It attaches.

7. the manufacturing method of cell attachment device according to claim 6, it is characterised in that: formed the cell adsorption layer it Before, further include having to form a laminated magnetic film on the substrate, there is a patterning, and forming the magnetism warming layer When, a plurality of specific position arrangements that there is magnetic warming macromolecule to correspond in the patterning.

8. the manufacturing method of cell attachment device according to claim 6, it is characterised in that: further include having in the substrate Side has the permanent magnet of a specific pattern, has a patterning, and when forming the magnetism warming layer, this is a plurality of Warming macromolecule with magnetic particle is arranged along the profile of the patterning.

9. the manufacturing method of cell attachment device according to claim 7 or 8, it is characterised in that: the object features are temperature The combination of degree and magnetic field, the cell attachment device a plurality of magnetic warming macromolecules in first temperature are contraction state, So that cell is adsorbed on the cell adsorption layer, in a second temperature, by the effect of the changes of magnetic field, so that only right Should the warming macromolecule in patterning maintain contraction state, allow cell mutually should patterning specific region On by the cell adsorption layer adsorb.

10. the manufacturing method of cell attachment device according to claim 7 or 8, it is characterised in that: further include being formed again Several second warming macromolecules, and when forming the magnetism warming layer, the magnetism warming macromolecule and second temperature will be contained Feel macromolecule mixed solution, be coated on the cell adsorption layer, so that there is the magnetism temperature on the region of the corresponding patterning Sense macromolecule is formed on the cell adsorption layer, and on other regions, there is the second warming macromolecule to be formed in the cell On adsorption layer.