A slide and tissue culture dish

Abstract

This invention relates to a tissue carrier, comprising: a carrier having at least one notch extending from the upper surface of the carrier to the lower surface of the carrier; and multiple support pillars, each having one end connected to the lower surface of the carrier; the multiple support pillars being evenly arranged around the axis of the carrier. Unlike existing technologies, both 0.5 cubic millimeter and 3 cubic millimeter tissue blocks can be clamped and fixed by the carrier fitting against the bottom of the culture chamber, thereby reducing the risk of displacement or detachment of the tissue blocks during medium changes or operations. The notch on the carrier facilitates cell migration. When replacing the complete culture medium subsequently, the medium in the culture chamber can be directly replaced through the notch on the carrier without removing the carrier, reducing the risk of tissue block contamination and enhancing the continuity of subsequent tissue block culture.

Description

Technical Field

[0001] This invention relates to the field of cell and tissue culture equipment technology, and particularly to a culture dish for a tissue block method. Background Technology

[0002] Tissue block culture, a classic and widely used technique, is based on the principle of allowing cells to naturally migrate and proliferate from the cut edge of a tissue block under in vitro culture conditions, thereby obtaining and expanding target cells, especially mesenchymal stem cells. This method is particularly suitable for culturing various mesenchymal stem cells, such as dental pulp stem cells, periodontal ligament stem cells, and adipose-derived stem cells, because it can maximally preserve the tissue microenvironment and intercellular interactions, which is crucial for maintaining the biological characteristics of stem cells.

[0003] Stem cells typically reside in specific tissue microenvironments, the so-called "stem cell niches," which are composed of the extracellular matrix, signaling molecules, and neighboring cells. These microenvironments subtly regulate stem cell self-renewal, multi-lineage differentiation potential, and quiescent state. The tissue block method, by directly attaching small pieces of tissue to a culture surface, effectively mimics the in vivo environment and avoids the cell membrane protein damage, stress responses, and phenotypic changes that can result from harsh treatments such as traditional enzymatic digestion (e.g., collagenase, trypsin), thus helping to maintain the pristine state of stem cells. During culture, cells migrate from the edges of the tissue block, a process involving complex cell movement and adhesion mechanisms. By optimizing culture conditions, selective stem cell proliferation can be promoted, and key characteristics can be maintained, such as the expression of specific mesenchymal stem cell surface markers and the preservation of multi-lineage differentiation capacity.

[0004] This method, due to its simplicity, low cost, and ability to obtain stem cells with high viability and stable genetic background, shows great potential in regenerative medicine and disease modeling. However, in practice, especially for certain tissues such as dental pulp and periodontal ligament tissues, which are relatively small in size, ensuring stable adhesion of tissue blocks during culture and preventing displacement or even detachment during medium changes or operations, thus affecting effective cell migration and the continuity of subsequent culture, remains a problem that needs attention. Summary of the Invention

[0005] To solve, or at least partially solve, the above-mentioned technical problems, the present invention provides a culture dish for a slide and tissue block method.

[0006] The present invention provides a carrier sheet, which includes a carrier sheet and a plurality of support columns. The carrier sheet has at least one notch that extends from the upper surface of the carrier sheet to the lower surface of the carrier sheet. One end of each of the plurality of support columns is connected to the lower surface of the carrier sheet. The plurality of support columns are evenly distributed around the axis of the carrier sheet.

[0007] Optionally, the slide has a circular structure; there are multiple notches, which are evenly distributed around the axis of the slide; the notches are arc-shaped.

[0008] Optionally, the lower surface of the slide is provided with a gelatin coating; and / or, the upper surface of the slide is provided with a gelatin coating.

[0009] Optionally, the carrier has multiple through holes, each of which extends from the upper surface of the carrier to the lower surface of the carrier; the multiple through holes are evenly distributed around the axis of the carrier.

[0010] Optionally, the height of the support column is 0.5mm to 3mm; the diameter of the end of the support column away from the carrier is greater than the diameter of the end of the support column close to the carrier.

[0011] The present invention provides a tissue block culture dish, which includes a base, a lid, and a slide as described in any of the preceding claims. The base has a culture chamber; the lid is disposed on the base and is used to cover the culture chamber; the slide is disposed in the culture chamber; and a sandwich layer for accommodating tissue blocks is formed between the slide and the bottom of the culture chamber.

[0012] Optionally, the bottom of the culture chamber is an inclined surface; the angle between the bottom of the culture chamber and the bottom surface of the chassis is 0.5°~2°.

[0013] Optionally, the bottom of the culture chamber has a liquid accumulation groove, which is located at a first position at the bottom of the culture chamber.

[0014] Optionally, the bottom of the culture chamber is provided with at least one set of limiting blocks, which includes two limiting blocks arranged opposite to each other, and a gap between the two limiting blocks is provided for a support column that accommodates the substrate.

[0015] Optionally, the bottom of the culture chamber has multiple sets of fixing protrusions, each set of fixing protrusions including multiple fixing protrusions, and the multiple fixing protrusions are evenly arranged in a circular shape.

[0016] Optionally, the cover has a receiving cavity with a diameter larger than the outer diameter of the chassis; the top of the chassis is disposed within the receiving cavity to cover the culture chamber.

[0017] Unlike existing technologies, this method allows for the clamping and fixation of tissue blocks, whether 0.5 cubic millimeters or 3 cubic millimeters in size, by fitting the carrier plate to the bottom of the culture chamber. This reduces the risk of displacement or detachment during medium changes or other procedures. The notch in the carrier plate facilitates cell migration. When replacing the complete culture medium, it can be directly replaced through the notch in the carrier plate without removing the carrier plate, reducing the risk of tissue block contamination and enhancing the continuity of subsequent tissue block cultures. Attached Figure Description

[0018] To more clearly illustrate the embodiments of the present invention, the relevant accompanying drawings will be briefly described below. It should be understood that the drawings described below are only for illustrating some embodiments of the present invention, and those skilled in the art can obtain many other technical features and connections not mentioned herein based on these drawings.

[0019] Figure 1 This is a schematic diagram of one embodiment of a carrier sheet of the present invention. Figure 1 ; Figure 2 This is a schematic diagram of one embodiment of a carrier sheet of the present invention. Figure 2 ; Figure 3 This is a schematic diagram of one embodiment of a carrier sheet of the present invention. Figure 3 ; Figure 4 This is an exploded view of an embodiment of a tissue block culture dish according to the present invention; Figure 5 This is a cross-sectional schematic diagram of an embodiment of a tissue block culture dish according to the present invention; Figure 6 This is a schematic diagram of an embodiment of the base of a tissue block culture dish according to the present invention. Figure 1 ; Figure 7 This is a schematic diagram of an embodiment of the base of a tissue block culture dish according to the present invention. Figure 2 ; Figure 8 This is a cross-sectional schematic diagram of an embodiment of the base of a tissue block culture dish according to the present invention.

[0020] Explanation of reference numerals in the attached figures: 1. Carrier; 11. Carrier; 12. Support column; 13. Notch; 15. Through hole; 2. Base plate; 21. Culture chamber; 22. Interlayer; 23. Liquid collection tank; 24. Limiting block; 25. Gap; 26. Fixing protrusion; 27. Ventilation groove; 3. Cover; 31. Receiving cavity. Detailed Implementation

[0021] To provide a more detailed understanding of the features and technical content of the embodiments of this disclosure, the implementation of the embodiments of this disclosure will be described in detail below with reference to the accompanying drawings. The accompanying drawings are for illustrative purposes only and are not intended to limit the embodiments of this disclosure. In the following technical description, for ease of explanation, several details are used to provide a full understanding of the disclosed embodiments. However, one or more embodiments may still be implemented without these details. In other cases, well-known structures and devices may be simplified in their depiction to simplify the drawings.

[0022] The terms "first," "second," etc., used in the specification, claims, and accompanying drawings of this disclosure are used to distinguish similar objects and are not necessarily used to describe a specific order or sequence. It should be understood that such data can be interchanged where appropriate for the embodiments of this disclosure described herein. Furthermore, the terms "comprising" and "having," and any variations thereof, are intended to cover non-exclusive inclusion.

[0023] In this disclosure, the terms "upper," "lower," "inner," "middle," "outer," "front," and "rear," etc., indicate the orientation or positional relationship based on the orientation or positional relationship shown in the accompanying drawings. These terms are primarily for better description of the embodiments of this disclosure and their implementations, and are not intended to limit the indicated devices, elements, or components to having a specific orientation, or to require them to be constructed and operated in a specific orientation. Furthermore, some of the aforementioned terms may be used to indicate other meanings besides orientation or positional relationship; for example, the term "upper" may in some cases indicate a dependency or connection relationship. Those skilled in the art can understand the specific meaning of these terms in the embodiments of this disclosure according to the specific circumstances.

[0024] Furthermore, the terms "set up," "connect," and "fix" should be interpreted broadly. For example, "connection" can be a fixed connection, a detachable connection, or an integral structure; it can be a mechanical connection or an electrical connection; it can be a direct connection or an indirect connection through an intermediate medium, or it can be an internal connection between two devices, components, or parts. Those skilled in the art can understand the specific meaning of the above terms in the embodiments of this disclosure according to the specific circumstances.

[0025] Unless otherwise stated, the term "multiple" means two or more, and "multiple groups" means two or more groups.

[0026] It should be noted that, unless otherwise specified, the embodiments and features described in the present disclosure can be combined with each other.

[0027] The inventors discovered that relatively small tissue blocks such as dental pulp and periodontal ligament may shift or even detach during culture, affecting cell migration and the continuity of subsequent culture. Furthermore, existing culture dishes may not facilitate medium changes, potentially leading to tissue block contamination.

[0028] In view of this, the inventors of the present invention provide a culture dish for a tissue block method to solve the above problems. Several specific embodiments of the present invention will be described in detail below with reference to the accompanying drawings.

[0029] First Embodiment The carrier sheet mentioned in this embodiment, such as Figure 1 , Figure 2 As shown, the slide includes a slide 11 and multiple support pillars 12. Specifically, a notch 13 with an arc shape is provided at the edge of the slide 11. The notch 13 extends from the upper surface of the slide 11 to the lower surface of the slide 11, i.e., the notch 13 penetrates the slide 11. Multiple support pillars 12 are all disposed on the lower surface of the slide 11, with one end of each pillar fixedly connected to the lower surface of the slide 11. The notch 13 is located between two adjacent support pillars 12. Providing a notch 13 on the slide 11 not only reduces the weight of the slide but also facilitates cell migration. Unlike existing technologies, the notch 13 on the slide 11 breaks the closed nature of the existing slide edge. When changing the complete culture medium, the presence of the notch 13 can be considered a pressure buffer channel, balancing the fluid pressure of the complete culture medium, thereby reducing the risk of tissue block displacement due to fluid pressure.

[0030] Optional, such as Figure 2 As shown, the support column 12 and the carrier plate 11 can be manufactured in one piece, for example, by injection molding to make them into a single component. This can avoid stress concentration at the connection interface and improve the mechanical strength of the overall structure.

[0031] Optionally, the support columns 12 can also be bonded to the lower surface of the slide 11 using a biocompatible adhesive. Multiple support columns 12 are evenly distributed around the axis of the slide 11. This even distribution ensures that the slide 11 is subjected to uniform force when placed on the bottom of the culture chamber of the tray, avoiding tilting of the slide due to unilateral settlement, and thus ensuring that the height of the interlayer formed between the slide 11 and the bottom of the culture chamber is consistent.

[0032] Optionally, based on the above scheme, the diameter of the end of the support column 12 closest to the slide 11 is smaller than the diameter of the end of the support column 12 furthest from the slide 11, that is, the support column 12 has a frustum-shaped structure. This structural design has multiple technical advantages. First, the larger diameter end of the support column 12 can increase the contact area between the support column 12 and the bottom of the culture chamber, allowing the slide 1 to be more stably placed in the culture chamber 21. Second, the frustum structure lowers the center of gravity of the support column 12, improving the overall anti-tipping ability of the slide 1. Optionally, the diameter of the end of the support column 12 furthest from the slide 11 is twice the diameter of the end of the support column 12 closest to the slide 11.

[0033] Optional, such as Figure 1 , Figure 2 As shown, the slide 11 has multiple notches 13 at its edge, and these notches 13 are evenly distributed around the axis of the slide 11. A notch 13 is provided between two adjacent support pillars 12. The multiple notches on the slide facilitate cell migration. They also further balance the fluid pressure of the complete culture medium, thereby reducing the risk of tissue displacement due to fluid pressure. By providing multiple notches on the slide, multiple channels for exchanging the complete culture medium are formed, allowing for faster replacement of the complete culture medium and preventing the accumulation of local metabolic waste.

[0034] In this embodiment, the arrangement of the carrier sheet 1 is as follows: Optionally, the slide 11 can be made of a transparent, biocompatible material. For example, slide 11 can be made of PC material or glass material. In use, researchers can directly observe the growth of the tissue block without moving slide 1.

[0035] Optionally, in this technical solution, the shape of the slide 11 is not limited, as long as the slide 11 can be placed in the culture chamber 21 of the tissue block culture dish.

[0036] Optional, such as Figure 1 As shown, the difference from the above technical solution lies in that the slide 11 has a circular shape. The diameter of the slide 11 is smaller than the diameter of the culture chamber of the tissue block culture dish, which facilitates the placement of the slide into the culture chamber. Multiple support columns 12 are evenly arranged around the axis of the slide 11, which facilitates the support columns to support the slide.

[0037] You can choose any one of the three technical solutions mentioned above. Of course, you can also combine several of them.

[0038] Second Embodiment In the first embodiment, the inventors discovered that there was a risk of tissue block displacement during fluid changes. Simultaneously, the inventors also found that the effective cell migration efficiency was relatively slow.

[0039] In view of this, this embodiment also proposes a carrier sheet. The second embodiment is a further improvement based on the first embodiment, the main improvement being: Optionally, gelatin is a biomaterial derived from collagen, possessing excellent cell adhesion. Applying gelatin to the lower surface of the slide 11, forming a gelatin coating, enhances the adhesion of the slide and the tissue block. Preferably, the thickness of the gelatin coating is between 10 and 50 micrometers to further enhance the adhesion of the tissue block and cells. If the thickness of the gelatin coating is less than 10 micrometers, there is a risk of tissue block and cell detachment. If the thickness of the gelatin coating is greater than 50 micrometers, it may affect the light transmittance of the slide.

[0040] Optionally, gelatin is applied to the upper surface of slide 11, and a gelatin coating is formed on the upper surface of slide 11 to facilitate cells to crawl from the lower surface of slide to the upper surface of slide.

[0041] Optionally, the two technical solutions mentioned above can be combined by applying a gelatin coating to the upper and lower surfaces of the slide 11. This arrangement can enhance the adhesion of the slide and facilitate the attachment of tissue blocks.

[0042] You may choose any one of the three technical solutions mentioned above.

[0043] Optional, such as Figure 3 As shown, the slide 11 has multiple through-holes 15, which extend from the upper surface to the lower surface of the slide 11, meaning all through-holes 15 penetrate the slide 11. When the slide 11 has a circular structure, the multiple through-holes 15 are evenly distributed around the axis of the slide 11. By providing through-holes on the slide, tissue blocks can easily grow from the lower surface of the slide to the upper surface through the through-holes. In this technical solution, through the coordinated work of the through-holes 15 and the notch 13, the notch 13 is mainly responsible for replacing the complete culture medium in the culture chamber, and of course, the notch is also responsible for cell migration. The through-holes 15 on the slide provide a physical channel for cells to cross the slide 11. Cells can start from tissue blocks on the lower surface of the slide 11, pass through the through-holes 15, and grow to the upper surface of the slide 11, thereby achieving true three-dimensional growth and increasing the cell yield per unit area.

[0044] Compared with existing technologies, the culture medium provided in this embodiment constructs a stable culture environment through the synergistic cooperation of notches, support pillars, gelatin coating, and through-holes. Support pillar 12 establishes physical space, preventing tissue blocks from being compressed. Notches 13 and through-holes 15 ensure unimpeded material exchange and balance fluid pressure. The gelatin coating provides adhesion. This combination of physical fixation and chemical adhesion, along with optimized fluid dynamics, effectively solves the technical problems of easy displacement and detachment of tissue blocks during culture.

[0045] Third Embodiment The tissue block culture dish mentioned in this embodiment, such as Figure 4 , Figure 5 As shown, the tissue block culture dish includes a base 2, a lid 3, and a substrate slide 1 as disclosed in the first or second embodiment. The base 2 has a culture chamber 21, which can be considered as the culture chamber 21 of the tissue block culture dish. The opening of the culture chamber 21 is located at the top of the base 2. The top of the base 2 has a venting groove 27, through which the culture chamber 21 communicates with the external environment. The lid 3 has a receiving cavity 31, the opening of which is located at the bottom of the lid 3. The lid 3 is detachably connected to the base 2. Specifically, the diameter of the receiving cavity 31 is larger than the outer diameter of the base 2. By placing the top of the base 2 inside the receiving cavity 31, the top of the base 2 abuts against the bottom of the receiving cavity 31, thus achieving a detachable connection between the lid 3 and the base 2. This also allows the lid 3 to cover the culture chamber 21 of the base 2, effectively preventing the risk of external dust and microbial aerosols from entering the culture chamber 21 from the side. When the lid 3 covers the base 2, the venting groove 27 is not sealed, but forms a gas channel between it and the lid. This gas channel allows air exchange and maintains a stable pH value in the culture environment. Preferably, there can be multiple venting grooves 27, which are evenly distributed around the axis of the base 2 to ensure the uniformity of gas concentration within the culture chamber 21. In use, the slide 1 is first placed in the culture chamber 21 of the base 2, with the support column 12 of the slide 1 abutting against the bottom of the culture chamber 21. At this time, a sandwich 22 for accommodating the tissue block is formed between the slide 11 of the slide 1 and the bottom of the culture chamber 21.

[0046] Optionally, not shown in the figure, the venting groove is S-shaped. Specifically, when the cover 3 covers the chassis 2, the venting groove 27 and the cover form a bent gas channel. This bent gas channel causes large dust particles and microbial aerosols in the external airflow to be deposited at the bend of the side wall of the venting groove due to inertial collision when passing through, and cannot enter the interior of the culture chamber 21.

[0047] Optionally, the height of the interlayer 22 is determined by the height of the support column 12. The structure of the support column 12 is improved, and its height ranges from 0.5mm to 3mm. Preferably, the height of the support column 12 is 0.5mm; or the height of the support column 12 is 1mm; or the height of the support column 12 is 1.5mm. The distance between the lower surface of the slide and the bottom of the culture chamber is the height of the interlayer 22. When the height of the support column 12 ranges from 0.5mm to 3mm, the height of the interlayer 22 also ranges from 0.5mm to 3mm. This arrangement facilitates the interlayer accommodating tissue blocks of different sizes. This interlayer space not only provides a physical containment area for the tissue blocks, preventing them from being flattened, but also forms a relatively static liquid layer, reducing the direct impact of the complete culture medium on the tissue blocks during medium changes.

[0048] Compared to existing technologies, this embodiment achieves not only physical sealing and protection through the systematic integration of a chassis with venting grooves, a substrate, and a lid, but also deeply optimizes gas exchange. The venting grooves balance the needs of sealing and ventilation, while the controlled design of the internal microenvironment further enhances the stability and success rate of cell culture. This synergy between the overall architecture and local details constitutes a highly integrated and fully functional tissue block culture system.

[0049] Fourth embodiment In the third embodiment, the inventors discovered that there is a risk of tissue block displacement during fluid exchange.

[0050] In view of this, this embodiment also proposes a tissue block culture dish. The fourth embodiment is a further improvement based on the third embodiment, mainly in the chassis, and the specific design is as follows: Optional, such as Figure 6 As shown, a set of limiting blocks 24 is provided on the chassis 2. This set of limiting blocks 24 includes two limiting blocks 24, which are arranged opposite each other and form a gap 25 between them. This gap 25 is used to accommodate the support column 12 of the slide 1. This fit strictly restricts the degree of freedom of the slide 1 in the horizontal direction. Even if it is subjected to impact or slight vibration when changing the complete culture medium, the slide 1 cannot move, thereby ensuring the spatial stability of the interlayer 22 and the fixation of the tissue block position.

[0051] Optional, such as Figure 6As shown, the chassis 2 is provided with multiple sets of limiting blocks 24, which are evenly distributed around the axis of the chassis 2 to provide all-round locking protection for the support column. Since the multiple sets of limiting blocks 24 have the same structure and arrangement, one set of limiting blocks 24 is used as an example for introduction. This set of limiting blocks 24 includes two limiting blocks 24, which are arranged opposite each other and form a gap 25 between them. This gap 25 is used to accommodate the support column 12 of the carrying plate 1.

[0052] Either of the two technical solutions mentioned above can be chosen.

[0053] Optional, such as Figure 6 As shown, the bottom of the culture chamber 21 is provided with multiple sets of fixing protrusions 26, which are spaced apart. Since the structure and arrangement of each set of fixing protrusions 26 are the same, one set is used as an example. This set of fixing protrusions 26 includes multiple fixing protrusions 26 arranged in a circle, and the multiple fixing protrusions 26 are evenly distributed around the center of this circle. That is, the multiple fixing protrusions are evenly distributed in a circular shape. In use, the tissue block can be placed between the multiple fixing protrusions so that the multiple fixing protrusions can limit and fix the tissue block. The height of the fixing protrusions 26 is less than the height of the support column 12, ensuring that they do not push up the carrier sheet 1, but are located within the space of the interlayer 22. In use, the tissue block can be precisely placed within the area surrounded by the multiple fixing protrusions 26. The fixing protrusions 26 constitute a physical fence at the microscopic level, preventing the tissue block from rolling or drifting before attachment.

[0054] Optional, such as Figure 3 , Figure 4 , Figure 6As shown, this technical solution is a further improvement on the technical solution with through holes 15 in the second embodiment. The bottom of the culture chamber 21 is provided with multiple sets of fixing protrusions 26, each set corresponding to a through hole 15. Each set of fixing protrusions 26 includes multiple fixing protrusions 26, which are evenly distributed around the axis of their corresponding through hole 15. In use, the tissue block can be placed between the multiple fixing protrusions to limit and fix the tissue block. The central axis around which each set of fixing protrusions 26 is located coincides with the axis of its corresponding through hole 15. When the slide 1 is placed into the culture chamber 21 and locked by the limiting block 24, a through hole 15 corresponds directly above the tissue block. This design confines the tissue block within a vertical columnar space formed by the bottom fixing protrusions 26 and the top through hole 15. After cells crawl out from the edge of the tissue block, they can either crawl out along the bottom of the culture chamber or grow upwards through the upper through hole 15. This structure not only enables the arraying and standardized arrangement of tissue blocks, facilitating subsequent automated microscopic scanning and image analysis, but also forces the construction of uniform three-dimensional growth channels, greatly improving the reproducibility of culture and the consistency of data. Each set of fixed protrusions can hold one tissue block, or multiple tissue blocks can be fixed. For example, multiple tissue blocks can be evenly distributed around the axis of the through-hole, with each tissue block located between two adjacent fixed protrusions.

[0055] Optionally, based on one of the two optional technical solutions mentioned above, the surface of the fixing protrusion 26 can also be coated with gelatin. This allows the tissue block to quickly acquire initial adhesion when it comes into contact with the fixing protrusion, further shortening the adhesion time.

[0056] You may choose any one of the three technical solutions mentioned above.

[0057] Of course, one of the two technical solutions with limit block 24 can be combined with one of the three technical solutions with fixed protrusion 26.

[0058] Fifth embodiment The inventors discovered that during culture medium replacement, there may be residual culture medium, resulting in incomplete replacement.

[0059] In view of this, this embodiment also proposes a tissue block culture dish. The fifth embodiment is a further improvement based on the third or fourth embodiment, with the main improvement being in the chassis, as detailed below: Optional, such as Figure 7 , Figure 8As shown, the bottom of the culture chamber 21 has a sloping structure, meaning that the bottom of the culture chamber 21 is inclined to the bottom of the base plate 2. Preferably, the angle α between the bottom of the culture chamber 21 and the bottom surface of the base plate 2 is 0.5°~2°. When changing the culture medium, the inclined bottom of the culture chamber facilitates the suction of the old culture medium by pipette. If the angle α is less than 0.5°, the guiding effect of gravity on the liquid is not significant, and some residual liquid of the complete culture medium may still remain at a higher position due to surface tension. If the angle α is greater than 2°, although the drainage effect is better, it will lead to uneven distribution of the complete culture medium depth in the culture chamber. This may cause tissue blocks at higher positions to be exposed above the complete culture medium surface and dry out, or cause the liquid layer at lower positions to be too deep, affecting the gas exchange efficiency.

[0060] Optional, such as Figure 7 , Figure 8 As shown, based on the above technical solution, the structure of the chassis 2 is further improved. A liquid accumulation groove 23 is provided at the bottom of the culture chamber 21, and the depth of the liquid accumulation groove 23 can be 0.5mm~1.5mm. Figure 8 In this design, the left end of culture chamber 21 is higher than the right end, and the sump 23 is located at the right end of culture chamber 21. Therefore, the sump 23 can be considered to be positioned at the lowest point of culture chamber 21, which is the first position of the sump 23 at the bottom of culture chamber 21. By placing the sump 23 at the bottom of the culture chamber, the old complete culture medium flows naturally to the lowest point along the slope under gravity during culture medium replacement and collects in the sump 23. With the pipette, the old culture medium flows into the sump along the inclined bottom of the culture chamber. This eliminates the need for multiple points of culture medium aspiration, making culture medium replacement easier. At this point, the operator only needs to insert the pipette tip into the sump 23 to aspirate all the old complete culture medium at once. This synergistic combination of sloping flow and low-point convergence significantly reduces the difficulty of changing the culture medium, reduces mechanical disturbance to the tissue block caused by repeated multi-angle aspiration, and also reduces the risk of fluctuations in the culture environment caused by the dilution of the new complete culture medium by residual old complete culture medium.

[0061] Either of the two optional technical solutions mentioned above can be chosen.

[0062] In this embodiment, the combination of the bottom of the culture chamber with the inclined structure and the liquid accumulation tank achieves high efficiency in liquid management, significantly improving the success rate, standardization and experimental efficiency of tissue block culture.

[0063] The exemplary usage process of the tissue block method culture dish disclosed in this embodiment is as follows: In this embodiment, the method of culturing target tissue (which is one of dental pulp stem cells, periodontal ligament stem cells, or adipose stem cells) using tissue block culture is used as an example.

[0064] like Figures 1 to 8 As shown, after obtaining the tissue blocks, they are evenly placed at the bottom of the culture chamber 21 of the base plate 2, with multiple tissue blocks arranged in an array and adjacent tissue blocks spaced 0.5cm to 1cm apart. The culture dish is slightly tilted to allow the tissue blocks to air dry for 1-2 minutes to enhance adhesion. During the drying process, the tissue blocks should always be kept moist. Then, the slide 1 is placed in the culture chamber 21, and the tissue blocks are clamped and fixed by the cooperation between the slide 1 and the bottom of the culture chamber 21. The complete culture medium (such as DMEM / F12 supplemented with 10% fetal bovine serum, 1% penicillin antibiotics and 10ng / ml bFGF) is preheated, and the preheated complete culture medium is injected into the culture chamber 21 through the notch 13 of the slide 1 and along the wall of the culture chamber 21 of the base plate 2 until the complete culture medium covers the tissue blocks. The base plate 2 is gently shaken to distribute the complete culture medium evenly, but care should be taken to avoid washing away the tissue blocks. Cover the culture chamber 21 with lid 3, thus covering it. The culture chamber 21 is ventilated to the outside environment through the venting groove 27. When changing the complete culture medium, simply use a pipette through the notch 13 of the slide 1 to remove the complete culture medium from the collection tank 23. This allows for the replacement of the complete culture medium without moving the tissue block.

[0065] Unlike existing technologies, this method allows for the clamping and fixation of tissue blocks, whether 0.5 cubic millimeters or 3 cubic millimeters in size, by fitting the carrier plate to the bottom of the culture chamber. This reduces the risk of displacement or detachment during medium changes or other procedures. The notch in the carrier plate facilitates cell migration. When replacing the complete culture medium, it can be directly replaced through the notch in the carrier plate without removing the carrier plate, reducing the risk of tissue block contamination and enhancing the continuity of subsequent tissue block cultures.

[0066] Finally, it should be noted that those skilled in the art will understand that many technical details have been presented in the embodiments of the present invention to facilitate a better understanding of the invention. However, even without these technical details and various variations and modifications based on the above embodiments, the technical solutions claimed in the claims of the present invention can be substantially achieved. Therefore, in practical applications, various changes in form and detail can be made to the above embodiments without departing from the spirit and scope of the present invention.

Claims

1. A carrier sheet, characterized in that, include: A slide having at least one notch extending from the upper surface of the slide to the lower surface of the slide; Multiple support columns, one end of which is connected to the lower surface of the substrate; the multiple support columns are evenly distributed around the axis of the substrate.

2. The carrier sheet according to claim 1, characterized in that, The slide has a circular structure; there are multiple notches, which are evenly distributed around the axis of the slide; the notches are arc-shaped.

3. The carrier sheet according to claim 1, characterized in that, The lower surface of the slide is provided with a gelatin coating; and / or, the upper surface of the slide is provided with a gelatin coating.

4. The carrier sheet according to claim 1, characterized in that, The carrier has multiple through holes, all of which extend from the upper surface of the carrier to the lower surface of the carrier. The multiple through holes are evenly distributed around the axis of the carrier.

5. The carrier sheet according to claim 1, characterized in that, The height of the support column is 0.5mm~3mm; The diameter of the end of the support column furthest from the substrate is greater than the diameter of the end of the support column closest to the substrate.

6. A tissue block culture dish, characterized in that, include: The chassis has a culture chamber; A lid is provided on the chassis and is used to cover the culture chamber; The slide as described in any one of claims 1 to 5 is disposed within the culture chamber; a space for accommodating tissue blocks is formed between the slide and the bottom of the culture chamber.

7. The tissue block culture dish according to claim 6, characterized in that, The bottom of the culture chamber is an inclined plane; the angle between the bottom of the culture chamber and the bottom surface of the base plate is 0.5°~2°.

8. The tissue block culture dish according to claim 7, characterized in that, The bottom of the culture chamber has a liquid accumulation groove, which is located at a first position at the bottom of the culture chamber.

9. The tissue block culture dish according to claim 6, characterized in that, The bottom of the culture chamber is provided with at least one set of limiting blocks, which includes two limiting blocks arranged opposite to each other, and there is a gap between the two limiting blocks for accommodating a support column for the substrate.

10. The tissue block culture dish according to claim 6, characterized in that, The bottom of the culture chamber has multiple sets of fixed protrusions, each set of fixed protrusions including multiple fixed protrusions, which are evenly arranged in a circular shape.

11. The tissue block culture dish according to claim 6, characterized in that, The lid has a receiving cavity. The diameter of the receiving cavity is larger than the outer diameter of the chassis; The top of the chassis is positioned inside the receiving cavity to cover the culture cavity.

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