KR101870948B1 - Method of fabricating 3D biomimetic tissue scaffold - Google Patents
Method of fabricating 3D biomimetic tissue scaffold Download PDFInfo
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- KR101870948B1 KR101870948B1 KR1020160019373A KR20160019373A KR101870948B1 KR 101870948 B1 KR101870948 B1 KR 101870948B1 KR 1020160019373 A KR1020160019373 A KR 1020160019373A KR 20160019373 A KR20160019373 A KR 20160019373A KR 101870948 B1 KR101870948 B1 KR 101870948B1
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Abstract
A three-dimensional organ model structure manufacturing method and a three-dimensional organ model structure are disclosed, wherein the method of manufacturing a three-dimensional organ model structure comprises the steps of: fabricating a first unremarkable mold having a first indented shape; Forming a first relief mold having a first embossed shape corresponding to the first relief shape, preparing a second relief mold having a second relief shape corresponding to the first relief shape using the relief mold, And forming an organ model structure having a second relief shape corresponding to the second relief pattern using the second relief mold.
Description
The present invention relates to a method for producing a structure in which living organs are three-dimensionally simulated, and a three-dimensional organs simulated structure produced thereby.
For food or drug development, previous clinical trials and clinical trials are the steps to predict and evaluate the hazard and efficacy of the ingredients. Although animal or human cells can be used to predict adverse effects and efficacy prior to animal testing and clinical testing, cell culture models are not predictive enough to eventually require animal testing and clinical trials, It takes a lot of development time.
Regarding the improvement of the cell culture model, it is reported that when cells are cultured in a cell culture scaffold similar to the tissue structure of a human body without culturing the cells in a general culture dish, cell activity and functionality are improved . Accordingly, there is an increasing need to develop a three-dimensional organs-like structure capable of culturing cells corresponding to a three-dimensional tissue structure.
The background technology of the present application is disclosed in Korean Patent Laid-Open Publication No. 2015-0126520 (Application No. 2014-0053585).
It is an object of the present invention to solve the problems of the prior art described above, and it is an object of the present invention to provide a method and apparatus for reproducing a tissue structure of a complex organs three-dimensionally, Dimensional long-term simulation structure and a three-dimensional long-term simulation structure that can accurately reproduce the same.
It should be understood, however, that the technical scope of the embodiments of the present invention is not limited to the above-described technical problems, and other technical problems may exist.
According to an aspect of the present invention, there is provided a method of manufacturing a three-dimensional organ model structure, including: (a) fabricating a first negative-type mold having a first indentation; (b) fabricating a positive embossed mold having a first relief shape corresponding to the first relief pattern using the first relief mold; (c) fabricating a second negative-tone mold having a second engraved shape corresponding to the first embossed shape using the positive mold; And (d) fabricating an organ model structure having a second relief shape corresponding to the second relief pattern using the second relief mold.
As a technical means for accomplishing the above technical object, the three-dimensional organ model structure according to one embodiment of the present invention is a three-dimensional organ model structure structure manufactured by the method of manufacturing a three-dimensional organ model structure according to one embodiment of the present invention .
The above-described task solution is merely exemplary and should not be construed as limiting the present disclosure. In addition to the exemplary embodiments described above, there may be additional embodiments in the drawings and the detailed description of the invention.
According to the above-mentioned task solution of the present invention, it is possible to reproduce the tissue structure of a complex organs three-dimensionally more accurately than in the prior art, and also to reproduce the above-mentioned accurate reproducibility through a cell-friendly material such as a hydrogel having soft properties Simulation) can be made possible. In this regard, it has been reported that the function of cells is improved by simply culturing the cells in a simple three-dimensional form. According to the task solution of the present invention, three-dimensional A long-term simulated structure can be provided, the physiological similarity can be further increased, and the function of the actual tissue can be reproduced with higher reproducibility.
In addition, according to the above-described task solution of the present invention, a three-dimensional organs-like structure capable of maintaining high compatibility with existing experimental techniques and experimental instruments can be manufactured. For example, an observable level of a three-dimensional organs mimetic structure can be produced by a general microscope used for cell observation.
In addition, according to the present invention, it is possible to use the screening platform for testing the efficacy and side effects of drugs, cosmetics, and food ingredients. For example, the three-dimensional organ model structure manufactured by the task solution of the present application can be utilized as an artificial field model for evaluating the water absorption rate of food ingredients. In addition, the three-dimensional organs simulated by the method of the present invention can be developed into an experimental model of a specific disease and utilized in medical research. For example, a three-dimensional organs-like structure produced by the task solution of the present invention can be manufactured into a model of intestinal inflammation disease and can be utilized in the development of therapeutic agents and the study of disease mechanisms. The three-dimensional organs simulated by the method of the present invention can be utilized for the development of customized therapeutic agents. For example, a disease model reflecting the genetic characteristics of an individual can be developed using a three-dimensional organ model structure manufactured by the task solution of the present invention, and thus an individual-optimized therapeutic agent and treatment method can be tested.
BRIEF DESCRIPTION OF THE DRAWINGS Figure 1 is an overall flow diagram of a method for fabricating a three dimensional organ model structure according to one embodiment of the present application.
2 is a detailed flowchart of one implementation of step S110 of the method for fabricating a three-dimensional organ model structure according to one embodiment of the present application.
3 is a conceptual diagram for explaining an embodiment of step S110 of the method of manufacturing a three-dimensional organ model structure according to an embodiment of the present invention.
4 is a conceptual diagram for explaining an embodiment of step S120 of the method for fabricating a 3D long-term simulation structure according to an embodiment of the present invention.
5 is a detailed flowchart of a first embodiment of step S130 of the method for manufacturing a three-dimensional organ model structure according to an embodiment of the present invention.
FIGS. 6 and 7 are conceptual diagrams for explaining a first embodiment of step S130 of the method for manufacturing a three-dimensional organ model structure according to an embodiment of the present invention.
FIG. 8 is a detailed flowchart of a first embodiment of step S140 of the method for manufacturing a three-dimensional organ model structure according to one embodiment of the present application.
9 is a conceptual diagram for explaining a first embodiment of step S140 of the method for manufacturing a three-dimensional organ model structure according to an embodiment of the present invention.
10 is a detailed flowchart of a second embodiment of the step S130 of the method for manufacturing a three-dimensional organ model structure according to the embodiment of the present application.
11 is a conceptual diagram for explaining a second embodiment of the step S130 of the method of manufacturing a three-dimensional organ model structure according to an embodiment of the present invention.
FIG. 12 is a schematic perspective view for explaining an example of a stamp unit used in a step S130 and a step S140 of the method of manufacturing a three-dimensional organ model structure according to an embodiment of the present invention.
13 is a detailed flowchart of a second embodiment of the step S140 of the method for manufacturing a three-dimensional organ model structure according to an embodiment of the present invention.
FIGS. 14 and 15 are conceptual diagrams for explaining a second embodiment of step S140 of the method for manufacturing a three-dimensional organ model structure according to an embodiment of the present invention.
FIG. 16 is a conceptual diagram for explaining one function of the master unit of the stamp unit used in the steps S130 and S140 of the method of manufacturing a three-dimensional organ model structure according to an embodiment of the present invention.
Hereinafter, embodiments of the present invention will be described in detail with reference to the accompanying drawings so that those skilled in the art can easily carry out the present invention. It should be understood, however, that the present invention may be embodied in many different forms and should not be construed as limited to the embodiments set forth herein. In the drawings, the same reference numbers are used throughout the specification to refer to the same or like parts.
Throughout this specification, when a part is referred to as being "connected" to another part, it is not limited to a case where it is "directly connected" but also includes the case where it is "electrically connected" do.
It will be appreciated that throughout the specification it will be understood that when a member is located on another member "top", "top", "under", "bottom" But also the case where there is another member between the two members as well as the case where they are in contact with each other.
Throughout this specification, when an element is referred to as "including " an element, it is understood that the element may include other elements as well, without departing from the other elements unless specifically stated otherwise.
The present invention relates to a method for producing a three-dimensional synovial structure and a three-dimensional synovial structure produced by the method. More specifically, the present application can be used to produce cell culture supports (three-dimensional organs) that are similar to those of human organs using cell-friendly materials (such as hydrogels such as gelatin hydrogel, collagen, etc.) .
The present invention can be applied to a method of constructing a structure in which villus existing on the intestinal wall is three-dimensionally simulated, but the present invention is not limited thereto. For example, according to the present application, other organ structures other than the intestinal lining structure can be produced. In addition to the above-mentioned hydrogel or collagen, a method of manufacturing a three-dimensional organ model structure using another material (another kind of hydrogel) may also be included in the present invention.
In addition, in the present invention, in order to fabricate a hydrogel for cell culture with a desired three-dimensional microstructure (for example, a size of several tens of micrometers), a semiconductor fine processing, a replica molding, a stamping method, And the like can be used in combination. In other words, the above processes can be combined in various ways in consideration of the shape and precision of the tissue to be manufactured by the present invention, material properties, cost, time, and the like.
According to the present invention, unlike the conventional method in which only a simple microstructure such as a right-angled shape can be realized, a three-dimensional long-term structure having various curves and aspect ratios can be manufactured and provided in a micrometer- have.
Hereinafter, a method for manufacturing a three-dimensional organ model structure according to one embodiment of the present invention (hereinafter referred to as a " method for manufacturing a three-dimensional organ model structure ") will be described.
BRIEF DESCRIPTION OF THE DRAWINGS Figure 1 is an overall flow diagram of a method for fabricating a three dimensional organ model structure according to one embodiment of the present application.
Referring to FIG. 1, the method of manufacturing a three-dimensional organ model structure (S100) includes steps S110 to S140.
Step S110 is a step of manufacturing the first negative-
FIG. 2 is a detailed flow chart of one embodiment of the step S110 of the method for manufacturing a three-dimensional organ model structure according to an embodiment of the present invention, FIG. 3 is a flowchart Fig. 8 is a conceptual diagram for explaining an embodiment. Fig.
Referring to FIG. 3 (c), the
Here, the three-dimensional solid shape is preferably understood as a concept opposite to a cylindrical shape having a constant width (diameter) along the depth direction. That is, the
Referring to FIGS. 2 and 3, step S110 of manufacturing the first negative-
Step S111 is a step of forming a
In step S112, a
In this case, step S112 is a first embodiment (and an exposure method) in which the three-dimensional solid shape is formed, wherein a groove or a hole-
Specifically, the first embodiment (and the exposure method) of step S112 is a method of making concave structures (see FIGS. 3B and 3C) by overexposing UV. For example, SU-8 2150 is applied to a
On the other hand, the step S112 is a second embodiment (exposure method using light refraction) in which the three-dimensional solid shape is formed. By arranging and exposing a diffuser for guiding light refraction on the
Specifically, the second embodiment of the step S112 (the exposure method using light refraction) is a method using an optical diffuser. For example, SU-850 is applied to a
The distance between the
Subsequently, SU-8 may be laminated one or more times on the crosslinked SU-8, if necessary, by repeating the above-described method. For example, when the UV is irradiated for 20 seconds after removing a diffuser (for example, an Opal glass diffuser), the three-dimensional solid shape may be formed into a three-dimensional solid shape having a constant width depending on the depth . At this time, it is preferable to draw a reference line in the
As described above, in steps S111 and S112, a groove or a hole-like three-dimensional solid shape varying in width according to the depth and a three-dimensional shape having no width variation according to the depth are connected to form a first indentation shape . ≪ / RTI >
Referring to FIGS. 2 and 3, after the photolithography process is performed according to the first embodiment or the second embodiment of the step S112, the
Illustratively, in step S113, PEB (Post Exposure Bake) was performed at 65 ° C for 5 minutes and at 95 ° C for 10 minutes, and developed using SU-8 Developer to remove unbridged SU-8 (Negative first angle mold) of the concave structure (a structure corresponding to a groove or a hole-like three-dimensional solid shape whose width changes according to the depth) can be manufactured.
Through the steps S111 to S113, the first negative-
On the other hand, the step S110 performed by the photolithography process described above can be replaced as necessary with the step of forming the first negative-
4 is a conceptual diagram for explaining an embodiment of step S120 of the method for fabricating a 3D long-term simulation structure according to an embodiment of the present invention.
4, in step S120, the
4, the Sylgard 184 A and B solutions are mixed at a ratio of 10: 1, and the mixed PDMS (Sylgard solution) is applied to the first negative-tone mold 1 (for example, an engraved SU- 0.0 > PMMA < / RTI > mold). Next, the pressure in the vacuum chamber is controlled to allow the PDMS to penetrate into the first negative-
Hereinafter, the first embodiment of steps S130 and S140, and the second embodiment of steps S130 and S140 will be described. The first embodiment may be a method of dissolving and separating a second negative mold (intermediate mold) from the manufactured long term simulation structure, and the second embodiment may be a method of manufacturing the long term simulation structure through stamping Method).
FIG. 5 is a detailed flow chart of a first embodiment of the step S 130 of the method for manufacturing a three-dimensional organ model structure according to an embodiment of the present invention, and FIGS. 6 and 7 are cross- And is a conceptual diagram for explaining the first embodiment of step S130.
5 to 7, step S130 of manufacturing the second negative-
6A, the frame including the
Next, as shown in Fig. 6 (b), the
Referring to FIG. 7A, in step S131, the
7 (a), the embossed
7B, in step S132, the
More specifically, for example, a polycarbonate membrane (8 μm / 25 mm) is covered on the second negative-
For reference, alginate has been mainly described as a material corresponding to the second negative-type mold (which may be referred to as an intermediate mold), but the present invention is not limited thereto. That is, it is possible to use a variety of gels if the gel can be melted under a physiological condition that is not harmful to the cells as the second negative mold (intermediate mold). For example, it is also possible to cross-link with agarose instead of alginate, then melt the cross-linked agarose gel by raising the temperature. Alternatively, it is also possible to cross-link a hydrogel such as collagen or gelatin, and then selectively melt through an enzyme to remove it. As a material corresponding to the second negative angle mold, a material such as collagen, gelatin, and agarose may be used. In the case of using a material other than alginate, Depending on the material, the method of crosslinking and melting afterwards may be different.
As understood from the material description of the second negative-angle mold (middle mold), the main purpose of the second negative-angle mold is to have a soft material such as hydrogel when separating the object (long-term simulation structure) from the second negative- So that the object can be separated in a direction to maintain the shape of the designed tissue structure perfectly without being deformed or broken. That is, as described above, the second negative-angle mold may be provided with a material (for example, a hydrogel such as alginate) that can be melted and separated to maintain the structure of the object as it is.
Next, referring to FIG. 7C, step S133 is a step S133 of separating the
FIG. 8 is a detailed flowchart of a first embodiment of step S140 of the method for manufacturing a three-dimensional organ model structure according to an embodiment of the present invention. FIG. 1 is a conceptual diagram for explaining a first embodiment.
8 and 9, step S140 of fabricating the
Step S141 is a step of injecting and crosslinking the material corresponding to the long
Specifically, for example, prepare DMEM, 1N NaOH and 8.23% collagen solution on ice, add 500 μl of 8.23% collagen, 50 μl of DMEM and 20 μl of 1N NaOH to the E-tube, give. The mixed collagen is injected flatly into a second negative mold (alginate mold) using a pipette. The mold injected with collagen solution is crosslinked in an incubator at 37 ° C for 20 to 30 minutes. 0.1% glutaraldehyde (in PBS, v / v) was sprayed onto the crosslinked collagen structure, followed by crosslinking for 3 hours at 37 ° C in an incubator.
Step S142 is a step of dissolving and removing the second negative-tone mold 3 (see Fig. 9 (b)).
Specifically, for example, when a crosslinked collagen structure is treated in a solution of 50 mM EDTA (in DI water, w / v) for 10 hours or more, the second negative mold (intermediate mold) Only the collagen structure remains. This collagen structure is treated in 7% L-glutamic acid (in 1N HCl, w / v) for 6 hours or more and washed three times a day with PBS. The collagen structure remaining after dissolving and removing the second negative-
FIG. 10 is a detailed flowchart of a second embodiment of the step S130 of the method for manufacturing a three-dimensional organ model structure according to an embodiment of the present invention. FIG. 12 is a conceptual diagram for explaining a second embodiment of the present invention. FIG. 12 shows an example of the stamp unit used in the steps S130 and S140 of the method for fabricating a three-dimensional organ model structure according to an embodiment of the present invention. And is a schematic three-dimensional diagram for explaining.
Referring to FIGS. 10 and 11, step S130 may include steps S131a and S132a as a second embodiment. For reference, the first embodiment of the above-described step S130 is a method using the second negative mold (intermediate mold) combined with the cover gasket, while the second embodiment of the step S130 uses the second negative mold Intermediate mold) can be used.
Step S131a is a step of pouring and hardening the silicone solution on the
Step S131a may be a step of manufacturing an engraved PDMS mold using an embossed PDMS mold. Illustratively, referring to FIG. 11 (a), Sylgard 184 A and B solution are mixed at a ratio of 10: 1, and then the mixed PDMS (Sylgard solution) is poured into the
More specifically, a relief PDMS is placed in a vacuum chamber, a predetermined amount of trichlorosilane is dropped on the slide glass, and the glass is put into a vacuum chamber. And a predetermined time is waited while a predetermined vacuum or higher is maintained by the pressure pump. When trichlorosilane coating is applied to the embossed PDMS mold, the intaglio PDMS can be well separated even after the PDMS is poured and hardened on the embossed PDMS mold
Step S132a is a step S132a of attaching the second negative-
Referring to FIG. 12, the
The height and detailed configuration of the
FIG. 13 is a detailed flowchart of a second embodiment of step S140 of the method for manufacturing a three-dimensional organ model structure according to an embodiment of the present invention. FIGS. 14 and 15 are views showing a method of manufacturing a three-dimensional organ model structure according to one embodiment of the present invention And is a conceptual diagram for explaining a second embodiment of the step S140. FIG. 16 is a conceptual diagram for explaining one function of the master unit of the stamp unit used in the steps S130 and S140 of the method for manufacturing a three-dimensional organ model structure according to an embodiment of the present invention.
Referring to FIGS. 13 to 15, step S140 may include steps S141a to S143a as a second embodiment.
The step S141a is a step of filling the hollow portion of the well-shaped unit 37 (insert well) having the upper opening therein with a material corresponding to the organ model structure 4 (see Fig. 14 (a)).
Illustratively, a
14, a
In addition, since the three-dimensional organ structure provided by the present invention can be said to be the main purpose of the cell culture, it is preferable that the present method for producing a three-dimensional organ structure (S100) is provided as a cell-friendly process in the first half thereof. For example, it is preferred that no organic solvents or high concentration of base or estimation conditions are applied to the cells. Accordingly, as a material corresponding to the
In step S142a, a
Step S142a may be performed in such a manner that the
Referring to FIGS. 12 and 14 (b), a protruding
The thickness of the
A technically important factor in the production of the
In step S142a, the projecting
Another important factor in mimicking the thickness of an organ, such as the small intestine in a human body, is the amount of material (such as hydrogel) corresponding to the
Step S143a is a step of incubating the material corresponding to the
Illustratively, a well unit 37 (Transwell 3470) with a
Next, the
FIG. 16 is a conceptual diagram for explaining one function of the master unit of the stamp unit used in the steps S130 and S140 of the method of manufacturing a three-dimensional organ model structure according to an embodiment of the present invention.
11 and 16, the
15 (a), in step S143a, when the material corresponding to the
16, when separating the
The pressure generating reduction action of the through-
As described above, according to the second embodiment (stamping method) of steps S130 and S140, the stamp unit stably fixes the microstructure (second relief shape) of the long-term simulation structure (hydrogel support) Can be faithfully reproduced.
In addition, the method for manufacturing a three-dimensional organ structure (S100) may include a step of culturing a cell on the organ model structure (4). Such a cell culturing step may be appropriately applied or applied to a known cell culturing method known to those of ordinary skill in the art to be simulated, so that a detailed description thereof will be omitted.
According to the present invention, it is possible to reproduce the tissue structure of a complex organs three-dimensionally more accurately than in the prior art, and it is also possible to reproduce the above-described precise cells through a cell-friendly substance such as a hydrogel having soft properties have. Also, according to the present application, a three-dimensional organs-like structure capable of maintaining high compatibility with existing experimental techniques and experimental instruments can be manufactured. For example, an observable level of a three-dimensional organs mimetic structure can be produced by a general microscope used for cell observation.
In addition, the present invention can provide a three-dimensional organ model structure manufactured by any one of various embodiments of the above-described three-dimensional organ model structure manufacturing method (S100).
Illustratively, the three-dimensional organ model structure may be a structure that simulates a field of the human body. More specifically, in the case of a human field, the height of the villi may be 0.5 mm to 1.6 mm, the average thickness of the duodenum and the plant may be about 1.5 mm, and the 3-dimensional organ structure of the present invention may be manufactured .
Further, as described above, the three-dimensional organs-like structure can be made of a cell-friendly material. Illustratively, the 3D long-term structure may be provided by preparing a desired shape of the hydrogel using a mold of various materials, but the material of the 3D long-term structure is not limited to a hydrogel, May be a material.
It will be understood by those of ordinary skill in the art that the foregoing description of the embodiments is for illustrative purposes and that those skilled in the art can easily modify the invention without departing from the spirit or essential characteristics thereof. It is therefore to be understood that the above-described embodiments are illustrative in all aspects and not restrictive. For example, each component described as a single entity may be distributed and implemented, and components described as being distributed may also be implemented in a combined form.
The scope of the present invention is defined by the appended claims rather than the detailed description, and all changes or modifications derived from the meaning and scope of the claims and their equivalents should be interpreted as being included in the scope of the present invention.
1: 1st negative angle mold
11: 1st indentation shape
12: Base member
13: Photoresist layer
14: Photomask
14a: pattern
2: Embossed mold
21: First embossed shape
22: bottom frame
23: middle frame
24: Top frame
25: Positive mold seating part
3: 2nd negative mold
31: 2nd engraved shape
32: Cover gasket
32a: hole of cover gasket
33: Membrane
34: Perforated plate
35: upper gasket
36: Stamp unit
36a:
However,
36c: Adhesive material
37: Well type unit
37a: upper circumference of the hollow portion
37b: permeable member
37c: Chemicals for crosslinking
4: long term simulated structure
41: second embossed shape
42: Base portion
Claims (18)
(a) fabricating a first negative-tone mold having a first engraved shape;
(b) fabricating a positive embossed mold having a first relief shape corresponding to the first relief pattern using the first relief mold;
(c) fabricating a second negative-tone mold having a second engraved shape corresponding to the first embossed shape using the positive mold; And
(d) fabricating an organ model structure having a second relief shape corresponding to the second relief shape using the second relief mold,
, ≪ / RTI &
The step (c)
(c1) pouring and hardening the silicone solution on the positive mold and separating the mold from the positive mold to produce the second negative mold; And
(c2) attaching the second negative-angle mold to the lower end of the stamp unit,
The step (d)
(d1) filling a hollow portion of a well-shaped unit having an upper portion opened with a material corresponding to the organ model structure;
(d2) stamping the material corresponding to the organ model structure with a second negative-shape mold by inserting a stamp unit with the second negative-angle mold on the hollow portion; And
(d3) incubating the organism corresponding to the organ model structure to produce the organ model structure, and then separating the stamp unit from the organ model structure.
Wherein the first indentation shape includes a three-dimensional solid shape of a groove or a hole shape whose width changes according to the depth,
Wherein in the step (a), the three-dimensional solid shape is formed by a photolithography process.
Wherein in the step (a), the three-dimensional solid shape is formed by using at least one of overexposure and exposure by light refraction.
The step (a)
(a1) forming a photoresist layer on the base member;
(a2) disposing and exposing a photomask having a pattern corresponding to the first intaglio shape on the photoresist layer to selectively crosslink the photoresist layer according to the pattern; And
(a3) developing the photoresist layer to remove the non-crosslinked portion to form the first negative-type mold having the first intaglio shape,
Wherein the method comprises the steps of:
Wherein the step (a2) is a step of exposing the photoresist layer to a crosslinking treatment corresponding to a three-dimensional solid shape of a groove or a hole shape varying in width depending on the depth, Gt;
In the step (a2), a diffuser for guiding light refraction is disposed on the photomask and exposed to light so that a bridge corresponding to a groove or a hole-like three-dimensional solid shape having a width varying in depth is formed Wherein the method comprises the steps of:
In the step (a1) and the step (a2), at least two of a three-dimensional solid shape of a groove or a hole shape varying in width depending on the depth and a three- And wherein the step of forming the three-dimensional organs is repeated.
In the step (b), the positive mold is manufactured by pouring and hardening a silicon solution on the first negative-angle mold, and then separating the first negative-angle mold from the first negative-angle mold.
A protrusion protruding in an outer radial direction is formed on an upper end of the stamp unit,
In the step (d2)
Wherein the projection is seated on an upper periphery of the hollow portion,
Wherein the base unit thickness of the organ model structure is determined by the height of the stamp unit and the thickness of the second negative-angle mold.
The second indentation shape of the second negative-tone mold is vertically penetrated,
Wherein the stamp unit is vertically formed to communicate with the second engraved shape.
In the step (d3)
Wherein when the stamp unit is detached from the organ model structure, gas is supplied into the second intaglio shape through the through-hole.
In the step (d3)
Wherein when the material corresponding to the organ model structure is incubated, chemicals for crosslinking the material corresponding to the organ model structure in the second intaglio shape are injected through the through-hole, .
(e) culturing the cells on the organ model construct.
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Food Science and Industry, (2012), Vol.45, No.3, pp 15-22. |
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