PILLAR ASSEMBLY AND PREPARING APPARATUS FOR SAMPLE BLOCK COMPRISING THE SAME

Abstract

The present invention relates to a pillar assembly and an apparatus for producing a sample block comprising the same, and according to one aspect of the present invention, it is possible to change arrangement of pillars, and sample slices can be easily obtained from the sample block.

Claims

1. A pillar assembly for use in an apparatus for producing a sample block, comprising a first base member having a plurality of through holes; a second base member mounted to said first base member; a plurality of pillars supported between said first and second base members and arranged to be exposed to the outside through each through hole; and a fixing member provided so as to detachably fix said first and second base members.

2. The pillar assembly according to claim 1, wherein the fixing member comprises at least one magnet for securing the first and second base members by magnetic force.

3. The pillar assembly according to claim 1, wherein a plurality of through holes comprise a plurality of first through holes which is located along the edge of the first base member and a plurality of second through holes which is located in the central area of the first base member, and the interval between two adjacent first through holes and the interval between two adjacent second through holes are different from each other.

4. The pillar assembly according to claim 1, wherein said pillar has smaller diameter as it is more distant from the through hole.

5. An apparatus for producing a sample block comprising a disk for supporting the bottom portion of the solidified material; a jig, which is seated on the disk, for supporting the side portions of the solidified material; and a pillar assembly that at least part of an area having the bio-matrix attached toward the top of the solidified material is inserted inside the jig, wherein said pillar assembly comprises a first base member having a plurality of through holes, which is placed so as to face said jig; a second base member mounted to said first base member; a plurality of pillars, for attaching the bio-matrix, which is supported between said first and second base members and arranged to be exposed to the outside through each through hole; and a fixing member provided so as to detachably fix said first and second base members.

6. The apparatus for producing a sample block according to claim 5, wherein a plurality of through holes comprise a plurality of first through holes which is located along the edge of the first base member and a plurality of second through holes which is located in the central area of the first base member, and the interval between two adjacent first through holes and the interval between two adjacent second through holes are different from each other.

7. The apparatus for producing a sample block according to claim 6, wherein the pillars are arranged in the first through holes in a process of immersing the pillars having the bio-matrix attached in the culture solution, and the pillars are arranged in the second through holes in a process of being inserted into the jig.

8. The apparatus for producing a sample block according to claim 5, wherein the jig comprises the main body to provide a space portion for holding the solidified material and at least one alignment rod which is extended from said main body and inserted into the pillar assembly in order to align the pillar assembly in a predetermined position on the jig.

9. The apparatus for producing a sample block according to claim 5, wherein the fixing member comprises at least one magnet for securing the first and second base members by magnetic force.

10. The apparatus for producing a sample block according to claim 5, wherein said pillar has smaller diameter as it is more distant from the through hole.

11. The apparatus for producing a sample block according to claim 5, wherein said solidified material is a frozen tissue embedding agent (OCT compound).

12. The apparatus for producing a sample block according to claim 5, wherein said pillar assembly and jig each are detachably provided from the solidified material being solidified.

13. An apparatus for producing a sample block comprising a thermally conductive pad for supporting the bottom portion of the solidified material; a disc on which said thermally conductive pad is seated; a jig, which is seated on the disk, for supporting the side portions of the solidified material; and a pillar assembly that at least part of an area having the bio-matrix attached toward the top of the solidified material is inserted inside the jig, wherein said pillar assembly comprises a first base member having a plurality of through holes, which is placed so as to face said jig; a second base member mounted to said first base member; a plurality of pillars, for attaching the bio-matrix, which is supported between said first and second base members and arranged to be exposed to the outside through each through hole; and a fixing member provided so as to detachably fix said first and second base members.

14. The apparatus for producing a sample block according to claim 13, wherein a plurality of through holes comprise a plurality of first through holes which is located along the edge of the first base member and a plurality of second through holes which is located in the central area of the first base member, and the interval between two adjacent first through holes and the interval between two adjacent second through holes are different from each other.

15. The apparatus for producing a sample block according to claim 14, wherein the pillars are arranged in the first through holes in a process of immersing the pillars having the bio-matrix attached in the culture solution, and the pillars are arranged in the second through holes in a process of being inserted into the jig.

16. The apparatus for producing a sample block according to claim 13, wherein the jig comprises the main body to provide a space portion for holding the solidified material and at least one alignment rod which is extended from said main body and inserted into the pillar assembly in order to align the pillar assembly in a predetermined position on the jig.

17. The apparatus for producing a sample block according to claim 13, wherein the fixing member comprises at least one magnet for securing the first and second base members by magnetic force.

18. The apparatus for producing a sample block according to claim 13, wherein said pillar has smaller diameter as it is more distant from the through hole.

19. The apparatus for producing a sample block according to claim 13, wherein said solidified material comprises paraffin.

20. The apparatus for producing a sample block according to claim 13, the disk is formed of a PDMS material, and the thermally conductive pad is formed of a metallic material.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0017] FIG. 1 is an exploded perspective view showing the pillar assembly related to one example of the present invention.

[0018] FIG. 2 is a plan view in the assembled state of the pillar assembly illustrated in FIG. 1.

[0019] FIG. 3 is a cross-sectional view in the assembled state of the pillar assembly illustrated in FIG. 1.

[0020] FIG. 4 is a perspective view for explaining one usage state of the pillar assembly illustrated in FIG. 1.

[0021] FIG. 5 is a cross-sectional view in the combined state of each component of FIG. 4.

[0022] FIG. 6 is a side view showing a pillar.

[0023] FIG. 7 is an exploded perspective view showing an apparatus for producing a sample block related to one example of the present invention.

[0024] FIG. 8 is a cross-sectional view in the combined state of each component of FIG. 7.

[0025] FIGS. 9 and 10 are cross-sectional views for explaining one usage state of the apparatus for producing a sample block illustrated in FIG. 7.

[0026] FIG. 11 is a plan view of an apparatus for producing a sample block related to another example of the present invention.

[0027] FIG. 12 is a photograph showing the apparatus for producing a sample block illustrated in FIG. 11.

DETAILED DESCRIPTION

[0028] Hereinafter, the pillar assembly and the apparatus for producing a sample block comprising the same according to one example of the present invention are explained in detail with reference to the attached drawings.

[0029] In addition, regardless of the reference numerals, the same or similar reference numerals are given to the same or corresponding components, for which redundant description will be omitted, and the size and shape of each illustrated structural member can be exaggerated or reduced for convenience of description.

[0030] FIG. 1 is an exploded perspective view showing the pillar assembly (100) related to one example of the present invention, FIG. 2 is a plan view in the assembled state of the pillar assembly (100) illustrated in FIG. 1, and FIG. 3 is a cross-sectional view in the assembled state of the pillar assembly (100) illustrated in FIG. 1.

[0031] The pillar assembly (100) related to one example of the present invention may be a pillar assembly (100) used in an apparatus for producing a sample block. In addition, the pillar assembly (100) includes a first base member (110) and a second base member (120) and a plurality of pillars (130) and a fixing member (140). Specifically, the pillar assembly (100) includes a first base member (110) having a plurality of through holes (111, 112), and a second base member (120) mounted to the first base member (110), a plurality of pillars (130) supported between the first and second base members (110, 120) and arranged to be exposed to the outside through each through hole (111, 112) and a fixing member (140) provided so as to detachably fix the first and second base members (110, 120).

[0032] Also, the first base member (110) and the second base member (120) may be a plate having a predetermined thickness. Furthermore, the first base member (110) and the second base member (120) may be formed of a metallic material. When the first base member (110) and the second base member (120) are combined, a space portion may be provided between them for part of the pillar area (e.g., head portion) to be placed and supported. The space portion may be also provided to have the same thickness as one of the head portion (131) of the pillar (130).

[0033] Besides, the pillar (130) may include a head portion (131) and a rod portion (132) extending from the head portion (131). The head portion (131) may be provided to have a diameter larger than one of the rod portion (132). The head portion (131) of the pillar (130) is placed in a space between the first base member (110) and the second base member (120) in a state that the first base member (110) and the second base member (120) combine. Furthermore, the rod portion (132) of the pillar (130) passes through the through holes (111, 112) of the first base member (110) to be exposed to the outside. Moreover, the bio-matrix, which will be described later, and cells are attached to the surface of the rod portion (132) of the pillar (130).

[0034] In addition, the length and diameter, and the like of the pillar (130) may be determined in various ways depending on the size and type of the sample block. In one example, a full length of the rod portion (132) of the pillar (130) may be about 7.1 mm, and the length of the rod portion (132) exposed to the outside of the first base member (110) may be about 4.1 mm. Furthermore, the diameter of the rod portion (132) may be about 2 mm, and the total thickness of the pillar assembly (100) in the state having the placed pillar may be about 12.1 mm.

[0035] The fixing member (140) may include one or more magnets (140) for securing the first and second base members (110, 120) by magnetic force. The magnet (140) may be provided on the first or second base members (110, 120). In addition, a plurality of magnets (140) may be arranged apart at a certain interval along a circumferential direction of the first or second base member (110, 120). For example, referring to FIG. 2, a plurality of magnets (140) may be provided on only the second base member (120), and each magnet (140) attached to the first base member (110) from a metal. Through such a structure, the first and second base members (110, 120) are removable. Furthermore, when the first and second base members (110, 120) are separated, the head portion (131) of the pillar (130) disposed between them is exposed to the outside. Moreover, each base member (120) may be also formed of a polymer material, and in this case, the magnets (140) may be provided on each base member (120) to be a pair with a different polarity.

[0036] A plurality of through holes (111, 112) may comprise a plurality of first through holes (111) which is located along the edge of the first base member (110), and a plurality of second through holes (112) which is located in the central region of the first base member (110). For example, the interval between two adjacent first through holes (111) and the interval between two adjacent second through holes (112) may be differently determined from each other. In addition, a plurality of first through holes (111) may be arranged in turn along a predetermined axial direction, and a plurality of second through holes (112) may be arranged in the central area to achieve a circle. Furthermore, the first through holes (111) may be formed the same as the well arrangement of a well plate that the pillar assembly (100) is immersed. For example, referring to FIG. 1, a plurality of first through holes (111) may be provided in a 5×5 lattice arrangement over the entire area from the edge side of the first base member (110) to the central area side. Moreover, a plurality of third through holes (121) may be provided on the second base member (120). At this time, each third through hole (121) is provided to have the same size as the first through hole (111). In addition, a plurality of third through holes (121) may have the same arrangement as one of a plurality of first through holes (111). For example, a plurality of third through holes (121) may be provided in a 5×5 lattice arrangement over the entire area from the edge side of the second base member (120) to the central area side. Furthermore, in a state that the first base member (110) and the second base member (120) combine, it is preferred that each first through hole (111) and each third through hole (121) corresponding thereto are coaxially arranged.

[0037] Meanwhile, in the cases of exposing the pillars (130) through a plurality of first through holes (111) to the outside and exposing the pillars (130) through a plurality of second through holes (112) to the outside, the arrangement of pillars may be different from each other. Accordingly, the arrangement of pillars may be easily changed depending on each process of producing the sample block.

[0038] FIG. 4 is a perspective view for explaining one usage state of the pillar assembly (100) illustrated in FIG. 1, FIG. 5 is a cross-sectional view in the combined state of each component of FIG. 4, and FIG. 6 is a side view showing a pillar (130).

[0039] First, in order to produce the sample block (20, see FIG. 8), the bio-matrix is attached to the surfaces of the pillars (130) and the pillars (130) of the pillar assembly (100) are immersed in the well plate (10) that a plurality of wells (well) containing a culture solution is arranged. For example, the well plate (10) may be a 96-well plate. At this time, cells may be 3-dimensionally cultured on the surfaces of the pillars (130) by immersing each pillar (130) in the wells (11). Then, in order to analyze the reactivity of drug in the cells, after reacting with a drug the cells 3-dimensionally cultured in the pillars (130), it can be transferred to the solidified material to form the sample block (20).

[0040] Meanwhile, the bio-matrix may be hereby collagen or alginate, and the cells may be cancer cells or stem cells. In addition, a candidate drug may be added to the culture solution in the well plate (10). Specifically, a “biological sample” may hereby include blood and other liquid samples of biological origin, biopsy specimens, solid tissue samples such as tissue cultures or cells derived therefrom, and more particularly, be tissues, extracts, cell lysates, whole blood, plasma, serum, saliva, ocular fluid, cerebrospinal fluid, sweat, urine, milk, ascitic fluid, synovial fluid, peritoneal fluid, and the like, and preferably cell samples, most preferably cancer cells or stem cells, but not limited thereto. The sample can be subjected to pretreatment prior to use, which may include, for example, filtration, distillation, extraction, concentration, inactivation of interfering components, addition of reagents, and the like. In addition, the “bio-matrix” is a substance that provides a space in which the cells are three-dimensionally cultured, and preferably may be bFGF (fibroblast growth factor)-alginate, heparin-alginate, polyglycolic acid, polyglycolic acid-collagen, laminin, matrigel, fibrin, collagen or alginate hydrogel, but is not limited thereto, as long as it has biocompatibility. Furthermore, the adhesive force between the surfaces of the pillars (130) and the bio-matrix can be further enhanced by treating the surfaces of the pillars (130) with any one compound selected from the group consisting of poly-L-lysine, poly-D-lysine, and dopamine and a compound containing a divalent metal ion such as Mg2+, Ca2+, Ba2+, and Cu2+.

[0041] In addition, the pillar (130) may have smaller diameter as it is more distant from the through hole (111). As described above, the pillar (130) includes a head portion (131) and a rod portion (132), wherein the rod portion (132) may be provided to have smaller diameter as at least part of an area is more distant from the through hole (111). That is, the diameter of the head portion (131) and the adjacent region (132a) (d1, for example, 2.22 mm) is formed higher than the diameter of the head portion (131) and the distant region (132b) (d2, for example, 1.98 mm). For example, the rod portion (132) may have a conical shape.

[0042] FIG. 7 is an exploded perspective view showing an apparatus for producing a sample block (200) related to one example of the present invention, FIG. 8 is a cross-sectional view in the combined state of each component of FIG. 7, and FIGS. 9 and 10 are cross-sectional views for explaining one usage state of the apparatus for producing a sample block illustrated in FIG. 7.

[0043] Referring to FIGS. 7 and 8, the apparatus for producing a sample block (200) comprises a disk (220) for supporting the bottom portion (B) of the solidified material (O), and a jig (210), which is seated on the disk (220), for supporting the side portions (S) of the solidified material (O) and a pillar assembly (100) that at least part of an area having the bio-matrix attached toward the top of the solidified material (O) is inserted inside the jig (210).

[0044] The pillar assembly (100) is the same as the pillar assembly (100) described through FIG. 1, and specifically, the pillar assembly (100) comprises a first base member (110) having a plurality of through holes (111, 112), which is placed so as to face the jig (210), and a second base member (120) mounted to the first base member (110), and a plurality of pillars (130), for attaching the bio-matrix, which is supported between the first and second base members (110, 120) and arranged to be exposed to the outside through each through hole (111, 112) and a fixing member (140) provided so as to detachably fix the first and second base members (110, 120).

[0045] In the apparatus for producing a sample block (200), the pillar assembly (100) is disposed for the rod portion (132) of the pillar (130) exposed to the outside of the first base member (110) to be inserted insider the jig (210). As described above, cells treated with a drug and 3-dimensionally cultured are attached via the bio-matrix to the rod portion (132).

[0046] In the sample block (20) for making the section block, cells (22) treated with a drug and 3-dimensionally cultured are inserted into the solidified material (cell fixing substance) (O) hold inside the jig (210) via the pillars (130). Then, if they are subjected to freezing in a state that the pillars (130) are inserted to complete the solidification, the sample block (20) is formed. Here, the solidified material (O) may be paraffin or a frozen tissue embedding agent (OCT compound).

[0047] As described above, a plurality of through holes (111, 112) may comprise a plurality of first through holes (111) which is located along the edge of the first base member (110), and a plurality of second through holes (112) which is located in the central area of the first base member (110). For example, the interval between two adjacent first through holes (111) and the interval between two adjacent second through holes (112) can be determined differently from each other. In addition, a plurality of first through holes (111) may be arranged in turn along a predetermined axial direction, and a plurality of second through holes (112) may be arranged in the central area to achieve a circle.

[0048] In the process (well plate immersion process) of attaching the bio-matrix to the pillars (130) and culturing the cells in three dimensions, that is, in the process of immersing the pillars having the attached bio-matrix in the culture solution, the pillars may be arranged in the first through holes (111). Unlike this, in the process of inserting them in the jig (210), the pillars (130) may be arranged in the second through holes (112). This is because the well arrangement in the well plate is different from the shape of the jig (210). Specifically, the shape of the jig (210) is subject to correspond to the shape of the disk (220). Therefore, it is also necessary to change the arrangement of the pillars (130), when the shape of the disc (220) is different from the shape that the wells are arranged in the well plate (10).

[0049] Meanwhile, the jig (210) may comprise the main body (212) to provide a space portion (211) for holding the solidified material (O) and at least one alignment rod (213) which is extended from the main body (212) and inserted into the pillar assembly (100) in order to align the pillar assembly (100) in a predetermined position on the jig (210). The main body (212) may have a cylindrical shape of the cylinder. Furthermore, the main body (212) may have a structure that the space portion (211) is opened to each of both sides. For example, referring to FIG. 8, when the space portion (211) is opened to each of vertical directions, the bottom portion of the space portion (211) may be closed by the disk (220), and the top portion of the space portion (211) may be closed by the pillar assembly (100). In addition, the alignment rod 213 may be numerously provided apart at a certain interval along a circumferential direction of the main body (212), and a plurality of alignment rods (213) each may be provided in a radial fashion along a circumferential direction of the main body (212). For example, the alignment rod (213) may be adapted to be inserted into the first through hole (111) of the first base member (110).

[0050] Meanwhile, referring to FIGS. 8 to 10, the pillar assembly (100) and the jig (210) may be detachably provided from the solidified material (20) to be solidified. The sample block (20) in a state that the pillar assembly (100) and the jig (210) each are separated comprises a solid layer (21) and a three-dimensional cell array (22) located within the solid layer (21). The 3-dimensional cell array (22) is formed by fixing the cells on the surfaces of the pillars (130) in the solid layer (21). Then, the sample slices (40) may be obtained by cutting the sample block (20) in a state that the jig (210) and the pillar assembly (100) are separated through a cutter (30). At this time, the pillar assembly (100) is separated to form holes (23) having a predetermined depth and the cutting of the sample block (20) is carried out from the floor surfaces of the holes (23), so that the sample slices (40) can be easily and accurately obtained.

[0051] FIG. 11 is a plan view showing an apparatus for producing a sample block related to another example of the present invention, and FIG. 12 is a photograph showing the apparatus for producing a sample block illustrated in FIG. 11.

[0052] Referring to FIGS. 11 and 12, the apparatus for producing a sample block can be used when the solidified material is in particular paraffin. Specifically, the apparatus for producing a sample block related to the present example comprises a thermally conductive pad (230) for supporting the bottom portion of the solidified material (paraffin), and a disk (240) on which the thermally conductive pad (230) is seated, and a jig, which is seated on the disk (240), for supporting the side portions of the solidified material and a pillar assembly that at least part of an area having the bio-matrix attached toward the top of the solidified material is inserted inside the jig.

[0053] At this time, the disk (240) may be formed of a PDMS (polydimethylsiloxane) material, and the thermally conductive pad (230) may be formed of a metallic material. That is, the shape and the material of the disk can vary depending on the kinds of the solidified material, and the thermally conductive pad may also be used additionally.

[0054] The preferred examples of the present invention as described above have been disclosed for illustrative purposes, those in the art having ordinary knowledge about the present invention will be possible to make various modifications, alterations, and additions within the spirit and scope of the present invention, and such modifications, alterations, and additions will be regarded as belonging to the claims below.

DESCRIPTION OF REFERENCES

[0055] 1. 100: Pillar assembly [0056] 2. 200: Apparatus for producing a sample block [0057] 3. 210: Jig [0058] 4. 220, 240: disc [0059] 5. 230: Thermally conductive pad