KR102514854B1 - 3d printer using bio-ink and microstructure building system including the same - Google Patents

Abstract

The microtissue construction system according to the present invention is a microtissue construction system that simulates solid cancer tissues for cancer progression research. It includes a 3D printer including a culture module for seating and culturing the 3D output formed by the 3D printer, and an injection device for injecting the classified spheroids into the 3D tumor microenvironment output transferred from the 3D printer.

Description

3D printer using bio-ink and microtissue building system including the same {3D PRINTER USING BIO-INK AND MICROSTRUCTURE BUILDING SYSTEM INCLUDING THE SAME}

The present invention relates to a 3D printer using bio-ink and a microtissue construction system including the same, and more particularly, to a rapid and uniform biotissue formation and drug or compound reactivity analysis using multi-layer 3D printing technology. , A 3D printer using bio-ink that can analyze the reactivity of spheroids at various times or in real time by quickly constructing an “environment similar to living tissue” in advance and culturing spheroids in the created environment to construct similar biological tissues And it relates to a microtissue building system including the same.

The conventional microtissue construction system has a disadvantage in that even if an anticancer drug is directly administered to cancer cells or an environment similar to a living tissue is configured to confirm the effect of directly acting on cancer cells, the process required for creating the environment takes a long time.

In addition, conventional spheroid injection devices are limited in drug compound reactivity analysis because they cannot classify and inject variously desired size spheroids when configuring a tumor microenvironment by culturing spheroids in an environment prepared by the above method. There is a problem that there is.

The present invention is to solve the above problems, and the reactivity of spheroids by culturing spheroids in an environment similar to the tissue constructed through a 3D printer and a 3D printer for rapid and uniform formation of similar biological tissues and analysis of drug or compound reactivity It is an object of the present invention to provide a 3D printer using bio-ink including an injection device capable of analyzing in various time periods or in real time, and a microtissue construction system including the same.

The tasks of the present invention are not limited to the tasks mentioned above, and other tasks not mentioned will be clearly understood by those skilled in the art from the description below.

In order to solve the above problems, according to one aspect of the present invention, the extrusion module is an extrusion module of a 3D printer for forming a three-dimensional output using bioink, and a first extrusion module for forming an inner layer by extruding a first material extrusion part; and a second extrusion unit extruding a second material to form an outer layer surrounding the inner layer.

Here, the extrusion module may first extrude the second material to form the lower surface of the outer layer, and simultaneously extrude the first material and the second material to form the side surface of the outer layer and the inner layer.

The first extrusion part. It may include a first hollow tube formed long in a direction in which the first material is extruded.

In addition, the first extrusion unit may include a first supply pipe connected to an upper portion of the first hollow tube so that the first material is supplied in a direction perpendicular to the extrusion direction of the first material.

The first supply pipe may include a plurality of unit supply members disposed spaced apart from each other at a predetermined interval above the first hollow pipe.

Here, in the unit supply member, a plurality of first flow passages arranged in the longitudinal direction of the first hollow tube may be respectively connected to the first hollow tube.

The second extrusion part may include a second hollow tube having a larger diameter than the first hollow tube of the first extrusion part.

The second extrusion unit may include a second supply pipe connected to the second hollow pipe with the first supply pipe therebetween so that the second material is supplied in a direction perpendicular to the extrusion direction of the second material. .

The second supply pipe may include a plurality of unit connection members arranged spaced apart from each other at predetermined intervals along the outer circumferential surface of the second hollow pipe so as to be connected to the upper portion of the second hollow pipe without interfering with the first supply pipe. there is.

At this time, the unit connection members may be spaced apart so that the first supply pipe can be disposed in a space between two adjacent unit connection members.

The inner layer and the outer layer may include different cells.

For example, the first material may be bioink containing fibroblasts.

Also, the second material may be bioink containing vascular endothelial cells.

In order to solve the above problems, according to another aspect of the present invention, a 3D printer using bio-ink is an extrusion module for forming a three-dimensional output product in which different cell layers are in contact with each other using bio-ink; And it may include a culture module for culturing the 3-dimensional output formed by the extrusion module.

The extrusion module may include: a first extrusion unit extruding the first bio-ink containing the first cells to form an inner cell layer; and a second extrusion part extruding the second bio-ink containing the second cells to form an outer cell layer surrounding the inner cell layer.

Here, the extrusion module extrudes the first bioink to form a lower surface of the outer cell layer, and simultaneously extrudes the first bioink and the second bioink to form a side surface of the outer cell layer and the inner cell layer. can do.

The first extrusion part. It may include a first hollow tube formed long in a direction in which the first bio-ink is extruded.

In addition, the first extruding unit may include a first supply pipe connected to an upper portion of the first hollow tube to supply the first bio-ink in a direction perpendicular to an extrusion direction of the first bio-ink.

The second extrusion part may include a second hollow tube having a larger diameter than the first hollow tube of the first extrusion part.

In addition, the second extrusion part has a second supply pipe connected to the upper part of the second hollow pipe with the first supply pipe therebetween so that the second bio-ink is supplied in a direction perpendicular to the extrusion direction of the second bio-ink. can include

The culture module includes a seating portion in which a seating space for seating the three-dimensional output product is formed; and a culture unit supporting the seating unit and supplying a culture medium to the seating space.

Here, the seating part is provided so that the outer circumferential surface of the extrusion module is in contact with the inner circumferential surface, and the end of the retracted extrusion module is provided to be spaced apart at a predetermined height from the seating surface on which the lower surface of the 3D output is seated, thereby forming the seating space. can

In the seating portion, an inlet groove may be formed so that the culture solution is supplied from the culture unit to the seating space.

The inlet groove may be formed toward the seating space on at least one of an outer circumferential surface and a lower surface of the seating portion.

The culture unit is provided so that the culture solution flows into the inner space, and the inflow groove is drawn into the inner space so that the received culture solution can be supplied to the seating space.

In the culture part, an open hole having a larger diameter than the diameter of the inner space is formed in the upper part of the inner space so that one side of the outer circumferential surface of the seating part may be fixed in contact with it.

In the culture module, a plurality of the seating parts and the culture parts corresponding thereto may be arranged in one direction.

The 3D printer using bio-ink as described above may further include a transfer module for transferring the culture module to an external injection space capable of injecting spheroids into the formed three-dimensional output object.

Here, the transport module may be provided so that the culture module can be moved in a direction perpendicular to the transport direction of the culture module.

In order to solve the above problems, according to another aspect of the present invention, the microtissue building system is a microtissue building system that simulates solid cancer tissue analogues for cancer progression research, wherein any one of claims 14 to 29 3D printer; and an injection device for receiving the cultured spheroids from the outside, classifying them according to size, and injecting the classified spheroids into the three-dimensional tumor microenvironment output transferred from the 3D printer.

At this time, the injection device, a supply module for collecting the cultured spheroids and classifying them according to size; and an injection module for injecting the spheroids classified by the supply module into the 3D tumor microenvironment output.

Here, the supply module is provided with an insertion space into which the plate accommodating the cultured spheroids is introduced, and turns the plate inserted into the insertion space from a first position to a second position and the cultured spheroids accommodated in the plate. a collection unit that collects into the collection area; And a classification unit connected to the collection unit to move the cultured spheroids collected in the collection area to a classification area and classifying the cultured spheroids moved to the classification area by size.

At this time, the plate may be provided to discharge the received spheroid through an open hole formed on one side.

The collection unit may be inserted into the insertion space so that the open hole of the plate faces upward in the first position.

In addition, the collection unit is rotated to the second position and connected to the classification unit, so that the cultured spheroids collected in the collection area can be delivered to the classification unit.

The classification unit may classify the spheroids by size by moving the cultured spheroids in one direction along the classification area.

The classification unit may provide a long classification area in a vertical direction from the bottom surface, and classify the cultured spheroids by size using gravity.

The classification unit includes a plurality of filter members having a plurality of holes and arranged in a plurality of directions along the longitudinal direction of the classification area, and the filter members are arranged such that the size of the holes gradually decreases along the moving direction of the cultured spheroids. can

According to the 3D printer using the bio-ink of the present invention and the microtissue construction system including the same, there is an effect that an environment similar to a living tissue can be rapidly constructed in advance using multi-layer 3D printing technology.

In addition, by culturing the spheroids in the above environment, there is an effect that the reactivity of the spheroids can be analyzed in various time periods or in real time by constructing a similar biological tissue.

The effects of the present invention are not limited to the effects mentioned above, and other effects not mentioned will be clearly understood by those skilled in the art from the description of the claims.

The above summary, as well as the detailed description of the preferred embodiments of the present application set forth below, will be better understood when read in conjunction with the accompanying drawings. Preferred embodiments are shown in the drawings for the purpose of illustrating the present invention. However, it should be understood that this application is not limited to the precise arrangements and instrumentalities shown.
1 is a view showing the overall appearance of a microstructure building system according to an embodiment of the present invention;
Figure 2 is a view showing a 3D printer of the microstructure building system according to an embodiment of the present invention;
Figure 3 is a view for explaining the extrusion module of the microstructure building system according to an embodiment of the present invention;
4 shows a first extrusion portion of a microstructure building system according to an embodiment of the present invention;
5 shows a second extrusion portion of a microstructure building system according to an embodiment of the present invention;
6 is a view for explaining a three-dimensional output of the microstructure building system according to an embodiment of the present invention;
Figure 7 is a view for explaining the culture module of the microtissue building system according to an embodiment of the present invention;
8 is a view for explaining a seating part of a microstructure building system according to an embodiment of the present invention;
9 is a view for explaining a state in which a plurality of culture modules of the microtissue building system according to an embodiment of the present invention are provided;
10 is a view for explaining an injection device of a microstructure building system according to an embodiment of the present invention;
11 is a view for explaining a collection unit of a microstructure building system according to an embodiment of the present invention;
12 is a view for explaining the rotation of the collection unit of the microstructure building system according to an embodiment of the present invention;
13 is a view for explaining a classification unit of a microstructure building system according to an embodiment of the present invention
14 is a view for explaining a filter member of a microstructure building system according to an embodiment of the present invention;
15 is a view for explaining a solid cancer tissue analogue for cancer progression research using a microtissue building system according to an embodiment of the present invention.

Hereinafter, a preferred embodiment of the present invention in which the object of the present invention can be realized in detail will be described with reference to the accompanying drawings. In describing the present embodiment, the same name and the same reference numeral are used for the same configuration, and additional description thereof will be omitted.

1 is a view showing the overall appearance of a microstructure building system according to an embodiment of the present invention, Figure 2 is a view showing a 3D printer of the microstructure building system according to an embodiment of the present invention, and FIG. It is a view for explaining the extrusion module of the microstructure building system according to an embodiment of the present invention, Figure 4 is a view showing the first extrusion part of the microstructure building system according to an embodiment of the present invention, Figure 5 is the present invention It is a view showing the second extrusion part of the microstructure building system according to an embodiment of the present invention, Figure 6 is a view for explaining a three-dimensional output of the microstructure building system according to an embodiment of the present invention, Figure 7 is the present invention It is a view for explaining the culture module of the microtissue building system according to an embodiment of the present invention, Figure 8 is a view for explaining the mounting portion of the microtissue building system according to an embodiment of the present invention, Figure 9 is a view of the present invention A view for explaining a state in which a plurality of culture modules of a microtissue building system according to an embodiment are provided, and FIG. 10 is a view for explaining an injection device of a microtissue building system according to an embodiment of the present invention. 11 is a view for explaining the collection unit of the microstructure building system according to an embodiment of the present invention, and FIG. 12 is a view for explaining the rotation of the collection unit of the microstructure building system according to an embodiment of the present invention. 13 is a view for explaining the classification unit of the microstructure building system according to an embodiment of the present invention, FIG. 14 is a view for explaining the filter member of the microstructure building system according to an embodiment of the present invention, and FIG. 15 is a diagram for explaining a solid cancer tissue analogue for cancer progression research of the microtissue building system according to an embodiment of the present invention.

As shown in FIG. 1, the microtissue building system 10 according to an embodiment of the present invention is a microtissue building system that simulates solid cancer tissue analogues for cancer progression research, and is largely a 3D printer (100) using bio-ink. ), the injection device 200 may be included.

In addition, the microtissue construction system 10 according to an embodiment of the present invention circulates a medium or buffer for simulating a solid cancer tissue analogue, and a circulation device for collecting used waste is mounted on the rear side, so that a long-term stable medium, You can supply a supply of drugs or compounds and collect used waste.

Specifically, the circulation device may be provided as a refrigerator composed of several chambers, and each chamber has an individual pump, so that the solution can be moved independently.

In addition, the circulation device may be connected to various equipment other than the above-described 3D printer 100 and injection device 200 by using a connector.

In addition, at least one of the above-described 3D printer 100 and injection device 200 may be further connected with an incubator device capable of sending a desired plate to the connected equipment using a barcode recognition technique.

For example, the incubator device may be provided so that the plate can be placed on or taken out from a designated shelf using a conveyor belt and a stepper motor.

In addition, the side of the incubator device is made of glass, so that the inside of the incubator can be checked, and a connecting door is formed so that a desired plate can be sent to the connected device.

Here, looking in detail at the 3D printer 100 using bio-ink, a three-dimensional output product can be formed using bio-ink.

Specifically, a three-dimensional output product in which different cell layers contact each other is formed using bio-ink, and then the formed three-dimensional output product may be placed and cultured.

The injection device 200 receives the cultured spheroids (S) from the outside and classifies them according to size, and the spheroids classified in the three-dimensional tumor microenvironment output transferred from the 3D printer 100 using the above-described bioink By injecting (S), it is possible to mimic solid cancer tissue analogues for cancer progression research.

A detailed look at the 3D printer 100 through FIG. 2 is as follows.

The 3D printer 100 using bio-ink may specifically include an extrusion module 120 and a culture module 140.

The extrusion module 120 may form a three-dimensional output product in which different cell layers contact each other using bio-ink, and the culture module 140 may place and culture the three-dimensional output product formed by the extrusion module 120. .

Therefore, the 3D printer 100 using bio-ink can form a 3D tumor microenvironment output by placing and culturing the 3D output formed by the extrusion module 120.

First, the extrusion module 120 may include a first extrusion unit 122 and a second extrusion unit, as shown in FIG. 3 .

Here, the first extrusion unit 122 extrudes the first material to form the inner layer (A), and the second extrusion unit 124 extrudes a second material different from the first material to wrap the above-described inner layer (A). An outer layer (B) may be formed.

Specifically, the extrusion module 120 extrudes the second material first to form the lower surface of the outer layer (B), and simultaneously extrudes the first material and the second material to form the side surface of the outer layer (B) and the inner layer (A). can form

In addition, the extrusion module 120 may be operated as described above, and then finally extrude the second material to form the outer layer (B) surrounding the entire inner layer (A).

In this case, the inner layer (A) and the outer layer (B) described in the present invention may include different cells, the first material is bioink containing fibroblasts, and the second material includes vascular endothelial cells. It may be a bioink that does.

Here, cell reactivity to a drug may be confirmed by replacing either one of the inner layer (A) and the outer layer (B) with a drug layer.

In addition, by replacing either the inner layer (A) or the outer layer (B) with the chemotaxis factor layer, it is possible to confirm cell mobility for the chemotaxis factor.

In addition, by forming only the outer layer (B) without forming the inner layer (A), it is also possible to form a cell culture body in the form of a hollow cylinder.

For example, when the second material of the outer layer (B) includes vascular endothelial cells, a structure similar to a blood vessel may be formed.

Next, looking in detail at the structure of the extrusion module 120 through Figures 3 to 5,

As shown in FIG. 3 , the first extrusion part 122 may include a first hollow tube 122a that is elongated in a direction in which the first material is extruded.

In addition, the first extrusion unit 122 includes a first supply pipe 122b connected to an upper portion of the first hollow tube 122a, so that the first material is supplied in a direction perpendicular to the extrusion direction of the first material, It is extruded through the hollow tube (122a) to form the above-described inner layer (A).

For example, the first supply pipe 122b may include a plurality of unit supply members 122ba disposed spaced apart from each other at predetermined intervals above the first hollow pipe 122a.

Here, in the unit supply member 122ba, a plurality of first flow passages 122ba1 having a length in the same direction as the longitudinal direction of the first hollow tube 122a may be arranged, and each of the plurality of first flow passages 122ba1 may have a first flow path 122ba1. Each may be connected to one hollow tube (122a).

Here, the first supply pipe 122b has a plurality of first flow passages 122ba1 arranged so that the amount of the first material supplied can be controlled. The number of unit supply members 122ba may be increased, or the number of first flow paths 122ba1 included in the unit supply member 122ba may be further increased.

Next, the second extrusion part 124 may include a second hollow tube 124a having a larger diameter than the first hollow tube 122a of the first extrusion part 122, as shown in FIG.

Here, if the second extrusion part 124 can form an outer layer (B) surrounding the inner layer (A), it will be said that even if the length, inner diameter and outer diameter are variously changed, they all fall within the scope of the present invention.

For example, the second hollow tube 124a may be provided to adjust the inner diameter in the direction of the central axis.

Furthermore, the second hollow tube 124a is provided to be stretched or contracted in the longitudinal direction, and the overall length of the second hollow tube 124a may be adjusted.

In addition, the second extrusion part 124 includes a second supply pipe 124b connected to the second hollow pipe 124a with the first supply pipe 122b interposed therebetween, in a direction perpendicular to the extrusion direction of the second material. As the second material may be supplied.

In other words, the second extrusion part 124 may be spaced apart from each other and connected so that a space in which the first supply pipe 122b can be disposed is provided between the second supply pipe 124b and the second hollow pipe 124a. If the inner layer (A) and the outer layer (B) can be formed by the first extruding part 122 and the second extruding part 124, the spaced distance and the connected form can be varied, all of which are within the scope of the present invention. will be said to belong to

However, for more detailed description, referring to FIG. 5 described above, the second supply pipe 124b includes a plurality of unit connecting members 124ba arranged spaced apart from each other by a predetermined distance along the outer circumferential surface of the second hollow pipe 124a. By including), the second material can be moved by being connected to the top of the second hollow tube 124a without interfering with the first supply tubes 122b arranged in plurality.

For example, the unit connection members 124ba may be spaced apart from each other so that the above-described first supply pipe 122b may be disposed in a space between two adjacent unit connection members 124ba.

Therefore, in the extrusion module 120, as shown in FIG. 6, the first extrusion unit 122 extrudes the first bio-ink containing the first cells to form an inner cell layer, that is, an inner layer A, By extruding the second bioink containing the second cells, an outer cell layer surrounding the inner cell layer, that is, the outer layer (B) may be formed.

Specifically, the extrusion module 120 may extrude the first bioink to form the lower surface of the outer cell layer, and simultaneously extrude the first bioink and the second bioink to form the side surface of the outer cell layer and the inner cell layer. As described above, only the second bio-ink is finally extruded so that the outer cell layer completely covers the inner cell layer.

Next, as shown in FIG. 7, the culturing module 140 for cultivating the three-dimensional output output from the extrusion module 120 through the seating space may include a seating unit 142 and a cultivating unit 144. can

Specifically, the seating portion 142 may form a seating space for seating a 3D output object.

The culture unit 144 may serve to support the above-described seating unit 142 and supply a culture medium to the seating space.

Referring to the shape of the seating portion 142 in more detail with reference to FIG. 8 , the seating portion 142 may be provided so that the outer circumferential surface of the extrusion module 120 is in contact with the inner circumferential surface.

At this time, the seating portion 142 is provided so that the end of the retracted extrusion module 120 is spaced apart at a predetermined height from the inner seating surface, so that the 3D output is seated at a predetermined distance from the end of the retracted extrusion module 120 towards the ground. A seating space in which the extruded three-dimensional output product is seated by the inner seating surface and the inner circumferential surface of the seating portion 142 may be formed.

At this time, the seating portion 142 may have an inlet groove 142a formed, and through this, the culture solution is moved from the culture unit 144 to the seating space, so that the culture solution can be supplied as a three-dimensional output.

Here, the inlet groove 142a may be formed toward the seating space on at least one of the outer circumferential surface and the lower surface of the seating portion 142 .

For example, as shown in the drawing, a plurality of inlet grooves 142a may be arranged along the outer circumferential surface of the seating portion 142, and a constant interval between adjacent inlet grooves 142a may be formed.

In addition, although not shown in the drawing, the inlet groove 142a is additionally formed from the outer lower surface of the seating portion 142 to the seating surface on which the 3-dimensional output product is seated, so that the amount of culture solution supplied can be adjusted. The formed inlet grooves 142a may be provided to open and close, respectively.

As shown in FIG. 7 described above, the culture unit 144 is provided so that the culture solution flows into the inner space, and the inflow groove 142a is inserted into the inner space so that the received culture solution can be supplied to the seating space.

In addition, in the culture unit 144, an open hole having a larger diameter than the diameter of the inner space is formed at the upper part of the inner space so that one side of the outer circumferential surface of the seating part 142 can be fixed in contact with it.

Specifically, the seating portion 142, based on the outer circumferential side on which the inlet groove 142a is formed, by providing a larger diameter on the outer circumferential side of the upper side, a step may be formed on the outside of the seating portion 142, as described above. As one step comes into contact with the open hole of the culture unit 144, the seating portion 142 can be stably fixed to the culture unit 144.

In addition, a plurality of steps may be provided by providing the seating portion 142 in a form in which the diameter gradually increases from the bottom to the top, and accordingly, it is stably fixed to the opening holes of the various culturing units 144 in accordance with the diameter size. You can also make arrangements for it.

Furthermore, the culturing module 140 as described above can cultivate a plurality of three-dimensional outputs by providing a seating unit 142 and a plurality of cultivating units 144 corresponding thereto in one direction, as shown in FIG. 9 . there is.

The 3D printer 100 using bio-ink having the configuration described above is a culture module 140 as an external injection space capable of injecting spheroids (S) into the 3D tumor microenvironment output formed by culturing the 3D output. ) may further include a transport module 160 for transporting.

Therefore, the 3D printer 100 described above can transfer the culture module 140 to the injection device 200 disposed in the external injection space.

For example, the transport module 160 may be provided in a conveyor belt manner so that the culture module 140 can be moved in a direction perpendicular to the transport direction of the culture module 140 .

Therefore, the transfer module 160 can move the belt in one direction to move the culture module 140 provided on the top to one side of the injection device 200, and move in a direction different from the one direction so that the above-described prepared culture module 140 The 3D printer 100 may be moved to one side of the device.

Meanwhile, the injection device 200 may largely include a supply module 220 and an injection module 240 as shown in FIG. 10 .

The supply module 220 may collect the cultured spheroids (S) and classify them according to size, and specifically receive the plate in which the spheroids (S) are cultured in the above-described incubator device, and spheroids (S) in the plate can be collected and sorted according to size.

At this time, the injection module 240 may inject the spheroids (S) classified by the above-described supply module 220 into the three-dimensional tumor microenvironment output formed in the culture module 140 moved by the above-described movement module. .

As shown in FIG. 11 , the supply module 220 may include a collection unit 222 and a classification unit 224 .

Collecting unit 222 may be provided with an insertion space into which the plate containing the cultured spheroids (S) is drawn.

In addition, the collection unit 222 may collect the cultured spheroids (S) accommodated in the plate into the collection area by inverting the plate inserted into the insertion space from the first position to the second position.

At this time, the classification unit 224 is connected to the collection unit 222 to move the cultured spheroids (S) collected in the collection area to the classification area 224a, and the cultured spheroids moved to the classification area 224a ( S) can be classified by size.

Here, the plate may have an open hole formed on one side so that the collection unit 222 can discharge the accommodated spheroids (S) only when the first position is turned over to the second position.

In other words, the collection unit 222, when the plate is inserted into the insertion space so that the open hole of the plate faces upwards in the first position, and then turned over to the second position, the open hole faces downward, Spheroids (S) can be accommodated.

For example, as shown in FIG. 12, the collection unit 222 is provided to be turned over from the first position to the second position through the rotation member 222b, and the collection area is provided in the form of a hopper 222a. It can be.

In addition, the collection unit 222 is provided to be connected to the classification unit 224 described above at the second position, so that the cultured spheroids (S) collected in the collection area in the form of a hopper 222a are connected to the classification unit 224 ) can be transmitted.

A connector member ( 222c) by vibrating after being mounted, it is also possible to smoothly pass the spheroids (S) collected in the collection area to the classification unit 224.

Next, the classification unit 224 may classify the spheroids (S) by size by moving the cultured spheroids (S) in one direction along the classification area (224a).

Therefore, in the injection module 240, it is possible to inject the spheroid (S) of a desired size into the three-dimensional tumor microenvironment output.

A detailed look at the configuration of the classification unit 224 through FIG. 13 is as follows.

The classification unit 224 may provide a classification area 224a long in a vertical direction from the bottom surface, and classify the cultured spheroids S by size using gravity.

Specifically, the classification unit 224 may include a filter member 224b.

The filter member 224b may have a plurality of holes and may be arranged in plurality along the longitudinal direction of the classification area 224a.

At this time, the filter member 224b is arranged so that the size of the hole gradually decreases along the moving direction of the cultured spheroids (S), so that the cultured spheroids (S) can be classified by size.

Specifically, as shown in FIG. 14, the filter member 224b has a space for accommodating the spheroids S that did not pass through the upper side based on the hole, and the spheroids S accommodated to the outside in this space The same flow path as the longitudinal direction may be formed so that the flow can flow out.

Therefore, by connecting a pump to the flow path, the accommodated spheroids (S) can be transferred to the outside and stored by size of spheroids (S), and spheroids (S) of the desired size among the spheroids (S) stored in the same way as described above can be sent to the injection module 240.

Therefore, as shown in FIG. 15, the injection module 240 injects spheroids (S) of a desired size into the three-dimensional tumor microenvironment output formed in the culture module 140 transferred from the 3D printer 100, so that cancer progresses. It will be possible to simulate solid tumor tissue analogues for research.

Therefore, the 3D printer using the bio-ink of the present invention and the microtissue construction system including the same can quickly configure an environment similar to a living tissue in advance using multi-layer 3D printing technology, and cultivate spheroids in the above environment. It is possible to analyze the reactivity of spheroids at various times or in real time by constructing a similar biological tissue.

As described above, the preferred embodiments according to the present invention have been reviewed, and the fact that the present invention can be embodied in other specific forms in addition to the above-described embodiments without departing from the spirit or scope is a matter of ordinary skill in the art. It is self-evident to them. Therefore, the embodiments described above are to be regarded as illustrative rather than restrictive, and thus the present invention is not limited to the above description, but may vary within the scope of the appended claims and their equivalents.

A: inner layer
B: outer layer
S: spheroid
P: plate
10: microtissue building system
100: 3D printer
120: extrusion module
122: first extrusion unit
122a: first hollow tube
122b: first supply pipe
122ba: unit supply member
122ba1: 1st euro
124: second extrusion part
124a: second hollow tube
124b: second supply pipe
124ba: unit connection member
140: culture module
142: seating part
142a: inlet groove
144: culture unit
160: transfer module
200: injection device
220: supply module
222: collection unit
222a: hopper
222b: rotation member
222c: connector member
224: classification unit
224a: classification area
224b: filter member
240: injection module

Claims (38)

As an extrusion module of a 3D printer to simulate solid cancer tissue analogues for cancer progression research,
a first extrusion unit extruding a first material, which is bioink containing fibroblasts, to form an inner layer; and
A second extrusion unit extruding a second material, which is bioink containing vascular endothelial cells, to form an outer layer surrounding the inner layer;
Characterized in that the second material is first extruded to form the lower surface of the outer layer, and the first material and the second material are simultaneously extruded to form the side surface of the outer layer and the inner layer,
The first extrusion part and the second extrusion part,
Characterized in that the total length is variable, and the diameter of the second extruded part is provided to be variable in correspondence with the diameter of the first extruded part, so that a solid cancer tissue-like body having at least one shape can be simulated,
extrusion module. delete According to claim 1,
The first extrusion part,
Characterized in that it comprises a first hollow tube formed elongated in the direction in which the first material is extruded,
extrusion module.
According to claim 3,
The first extrusion part,
Characterized in that it comprises a first supply pipe connected to the top of the first hollow tube so that the first material is supplied in a direction perpendicular to the extrusion direction of the first material,
extrusion module.
According to claim 4,
The first supply pipe,
Characterized in that it comprises a plurality of unit supply members arranged spaced apart from each other at a predetermined interval at the top of the first hollow tube,
extrusion module.
According to claim 5,
The unit supply member,
Characterized in that a plurality of first flow passages arranged in the longitudinal direction of the first hollow tube are respectively connected to the first hollow tube,
extrusion module.
According to claim 4,
The second extrusion part,
Characterized in that it comprises a second hollow tube of a larger diameter than the first hollow tube of the first extrusion part,
extrusion module.
According to claim 7,
The second extrusion part,
Characterized in that it comprises a second supply pipe connected to the second hollow pipe with the first supply pipe therebetween so that the second material is supplied in a direction perpendicular to the extrusion direction of the second material,
extrusion module.
According to claim 8,
The second supply pipe,
Characterized in that it comprises a plurality of unit connecting members arranged spaced apart from each other at a predetermined interval along the outer circumferential surface of the second hollow pipe so as to be connected to the upper portion of the second hollow pipe without interfering with the first supply pipe.
extrusion module.
According to claim 9,
The unit connecting member,
Characterized in that it is spaced apart so that the first supply pipe can be disposed in the space between two adjacent unit connection members,
extrusion module.
delete delete delete As a 3D printer to simulate solid cancer tissue analogues for cancer progression research,
An extrusion module for forming a three-dimensional output product in which different cell layers are in contact with each other using bio-ink; and
Including a culture module for seating and culturing the three-dimensional output formed by the extrusion module,
The extrusion module,
a first extrusion unit extruding a first material, which is bioink containing fibroblasts, to form an inner layer; and
A second extrusion unit extruding a second material, which is bioink containing vascular endothelial cells, to form an outer layer surrounding the inner layer;
Characterized in that the second material is first extruded to form the lower surface of the outer layer, and the first material and the second material are simultaneously extruded to form the side surface of the outer layer and the inner layer,
The first extrusion part and the second extrusion part,
Characterized in that the total length is variable, and the diameter of the second extruded part is provided to be variable in correspondence with the diameter of the first extruded part, so that a solid cancer tissue-like body having at least one shape can be simulated,
3D printer using bio-ink.
delete delete According to claim 14,
The first extrusion part,
Characterized in that it comprises a first hollow tube formed long in the direction in which the first bioink is extruded,
3D printer using bio-ink.
According to claim 17,
The first extrusion part,
Characterized in that it comprises a first supply pipe connected to the upper part of the first hollow pipe so that the first bio-ink is supplied in a direction perpendicular to the extrusion direction of the first bio-ink.
3D printer using bio-ink.
According to claim 18,
The second extrusion part,
Characterized in that it comprises a second hollow tube of a larger diameter than the first hollow tube of the first extrusion part,
3D printer using bio-ink.
According to claim 19,
The second extrusion part,
And a second supply pipe connected to the upper part of the second hollow pipe with the first supply pipe therebetween so that the second bio-ink is supplied in a direction perpendicular to the extrusion direction of the second bio-ink. ,
3D printer using bio-ink.
According to claim 14,
The culture module,
a seating portion in which a seating space for seating the three-dimensional output product is formed; and
Characterized in that it comprises a culture unit for supporting the seating unit and supplying a culture solution to the seating space,
3D printer using bio-ink.
According to claim 21,
the attachment part,
It is provided so that the outer circumferential surface of the extrusion module is in contact with the inner circumferential surface, and the end of the retracted extrusion module is provided to be spaced apart at a predetermined height from the seating surface on which the lower surface of the three-dimensional output is seated, thereby forming the seating space. Characterized in that,
3D printer using bio-ink.
The method of claim 22,
the attachment part,
Characterized in that an inlet groove is formed so that the culture solution is supplied from the culture unit to the seating space,
3D printer using bio-ink.
According to claim 23,
The inlet groove,
Characterized in that at least one of the outer circumferential surface and the lower surface of the seating portion is formed toward the seating space,
3D printer using bio-ink.
According to claim 23,
The culture department,
It is characterized in that the culture solution is provided to flow into the inner space, and the inlet groove is drawn into the inner space to supply the received culture solution to the seating space.
3D printer using bio-ink.
According to claim 25,
The culture department,
Characterized in that an open hole having a larger diameter than the diameter of the inner space is formed in the upper part of the inner space so that one side of the outer circumferential surface of the seating part is fixed in contact.
3D printer using bio-ink.
According to claim 21,
The culture module,
Characterized in that the seating portion and the corresponding culturing portion are arranged in a plurality in one direction,
3D printer using bio-ink.
According to claim 14,
Characterized in that it further comprises a transfer module for transferring the culture module to an external injection space capable of injecting spheroids into the formed three-dimensional output.
3D printer using bio-ink.
According to claim 28,
The transfer module,
Characterized in that the culture module is provided to be moved in a direction perpendicular to the transport direction of the culture module,
3D printer using bio-ink.
As a microtissue construction system that mimics solid cancer tissue analogues for cancer progression research,
The 3D printer according to any one of claims 14 and 17 to 29; and
Including an injection device for receiving the cultured spheroids from the outside, classifying them according to size, and injecting the classified spheroids into the three-dimensional output transferred from the 3D printer,
Microtissue Building System.
31. The method of claim 30,
The injection device,
A supply module for collecting the cultured spheroids and classifying them according to size; and
Including an injection module for injecting the spheroids classified by the supply module into the three-dimensional output,
Microtissue Building System.
According to claim 31,
The supply module,
An insertion space into which the plate accommodating the cultured spheroids is inserted, and the plate inserted into the insertion space is turned over from a first position to a second position to collect the cultured spheroids accommodated in the plate into a collection area collection department; and
Characterized in that it comprises a classification unit connected to the collection unit to move the cultured spheroids collected in the collection area to a classification area and classify the cultured spheroids moved to the classification area by size,
Microtissue Building System.
33. The method of claim 32,
The plate is
Characterized in that it is provided to discharge the spheroid accommodated through an open hole formed on one side,
Microtissue Building System.
34. The method of claim 33,
The collection unit,
In the first position, characterized in that the plate is inserted into the insertion space so that the open hole of the plate faces upward.
Microtissue Building System.
35. The method of claim 34,
The collection unit,
Characterized in that the cultured spheroids collected in the collection area are transferred to the classification unit by being rotated to the second position and connected to the classification unit,
Microtissue Building System.
33. The method of claim 32,
The classification unit,
Characterized in that by moving the cultured spheroids in one direction along the classification area to classify the spheroids by size,
Microtissue Building System.
37. The method of claim 36,
The classification unit,
Characterized in that the classification area is provided long in the vertical direction from the bottom surface, and the cultured spheroids are classified by size using gravity.
Microtissue Building System.
38. The method of claim 37,
The classification unit,
It has a plurality of holes and includes a plurality of filter members arranged along the longitudinal direction of the classification area, characterized in that the filter member is arranged so that the size of the holes gradually decreases along the moving direction of the cultured spheroid ,
Microtissue Building System.

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