A well plate for 3D cell culture

Abstract

The utility model is suitable for cell culture technical field provides a kind of well plate for 3D cell culture, including substrate, cell hole and culture medium hole are opened in the substrate, communication groove is opened between the cell hole and culture medium hole. In the utility model, only need to operate culture medium hole, cell inside cell and fixing glue keep stable, to significantly reduce cell loss risk, improve the repeatability and reliability of experiment, the flow channel is formed between the outer periphery of substrate and middle gap, and PBS buffer is filled therein, so that the outer periphery hole and the central hole are in the same culture environment, effectively alleviate the edge effect caused by temperature difference and evaporation, ensure the consistency of each hole culture condition, to improve the accuracy of overall experimental data, adopt " two hole punch through " structure, improve traditional 96 well plate to 192 hole design, while maintaining the convenience of operation greatly improve flux, suitable for high batch cell culture and detection, improve experimental efficiency.

Description

Technical Field

[0001] This invention belongs to the field of cell culture technology, and particularly relates to a well plate for 3D cell culture. Background Technology

[0002] Organoids are tissue analogs with a specific spatial structure, formed through three-dimensional (3D) in vitro culture of adult stem cells or pluripotent stem cells. They can structurally and functionally mimic real organs, maximally replicating in vivo tissue structure and function, and can be stably passaged long-term. Compared to traditional two-dimensional culture models, organoids represent an innovative technology that can encompass the entire physiological process of an organism, possessing advantages such as a closer resemblance to the composition and behavior of physiological cells, a more stable genome, and greater suitability for biological transfection and high-throughput screening. Compared to animal models, organoid models are simpler to operate and can be used to study the mechanisms of disease occurrence and development. Therefore, they have broad application prospects in organ development, precision medicine, regenerative medicine, drug screening, gene editing, and disease modeling. In 2013, organoids were named one of the top ten technologies of the year by *Science* magazine. In early 2018, organoids were named *Nature Methods*' Method of 2017.

[0003] Existing technologies include several utility model patents in the field of cell culture technology. Among them, utility model patent application number CN218521267U discloses a well plate for 3D cell culture. Its basic description is as follows: This utility model belongs to the field of 3D cell culture technology and discloses a well plate for 3D cell culture, including a culture medium plate. The culture medium plate has multiple conventional wells, and each of the conventional wells has an auxiliary hole on one side. The auxiliary hole is located on the culture medium plate and communicates with the conventional wells. Through this design, this utility model reduces the mechanical disturbance to the culture droplets during operations such as changing the medium and adding treatment reagents. This minimizes the differences in organoids cultured in each conventional well of the culture medium plate caused by interference during operation, facilitating subsequent standardized batch testing of organoids in the conventional wells and minimizing distortion of results due to operational differences.

[0004] Existing 3D cell culture plates are typically single-well structures, with cells and culture medium added to the same well simultaneously. When changing the culture medium, the operation must be performed directly in the well, which makes it easy to accidentally aspirate cells or cell fixation gel during the medium change process, thus affecting the accuracy and reproducibility of the experiment. In addition, the wells of conventional plates are isolated from each other, and the peripheral wells are more affected by environmental temperature differences and evaporation, which easily produces the "edge effect," resulting in differences in cell growth status between the peripheral wells and the central wells, affecting the uniformity of experimental data. On the other hand, traditional well plates mostly use the standard 96-well specification, which has limited throughput, and when used with automated equipment, they lack targeted limiting or fixing structures, making it difficult to achieve rapid and stable installation, thus limiting their application in high-throughput automated experiments. Utility Model Content

[0005] This invention provides a well plate for 3D cell culture, aiming to solve the problems of existing 3D cell culture well plates, which are usually single-well structures. Cells and culture medium are added to the same well at the same time. When changing the culture medium, the operation must be carried out directly in the well. It is very easy to accidentally aspirate cells or cell fixation gel during the medium change, which affects the accuracy and reproducibility of the experiment. In addition, conventional well plates are isolated from each other, and the peripheral wells are greatly affected by environmental temperature difference and evaporation, which easily produces the problem of "edge effect".

[0006] This invention is implemented as follows: a well plate for 3D cell culture includes: a substrate, in which cell wells and culture medium wells are formed, and a connecting groove is formed between the cell wells and the culture medium wells. The connecting groove is used to introduce buffer solution between the cell wells and the culture medium wells to reduce edge effects. A limiting part for cooperating with automated equipment is provided on the outer periphery of the substrate, and a fixing device is installed on both sides of the substrate.

[0007] Preferably, the volume of the culture medium wells is larger than that of the cell wells to provide more space for culture medium storage.

[0008] Preferably, the fixing device includes a connecting plate, and mounting blocks are symmetrically mounted on the outside of the connecting plate. The mounting blocks are fixedly connected to the base plate by bolts. The connecting plate has a protrusion, and a movable spring is mounted on the outer sleeve of the protrusion.

[0009] Preferably, the movable spring is externally fixedly connected to two connecting rods, one end of each connecting rod is fixedly connected to a telescopic rod, the telescopic ends of the two telescopic rods are fixedly connected to the same hanging rod, and a pull-back spring is provided on the outer sleeve of each telescopic rod.

[0010] Preferably, one end of the pull-back spring is fixedly connected to the outside of the connecting rod, and the other end of the pull-back spring is fixedly connected to the outside of the hanging rod.

[0011] Preferably, the hanging rod is provided with a hanging plate for fixing the hanging rod.

[0012] Compared with related technologies, the well plate for 3D cell culture provided by this utility model has the following beneficial effects:

[0013] 1. In this invention, cell wells and culture medium wells are arranged in pairs, with a connecting groove between them. This connecting groove allows the introduction of buffer solution between the cell wells and the culture medium wells, enabling nutrient transfer while avoiding direct disturbance to the cells. When changing the culture medium, only the culture medium wells need to be operated on, while the cells and fixative inside the cell wells remain stable, significantly reducing the risk of cell loss and improving the reproducibility and reliability of the experiment. This invention forms a flow channel between the outer periphery and the central gap of the substrate, which is filled with PBS buffer, ensuring that the peripheral wells and the central wells are in the same culture environment. This effectively alleviates the edge effect caused by temperature difference and evaporation, ensuring the consistency of culture conditions in each well, thereby improving the accuracy of the overall experimental data. The "two-well interconnection" structure improves the traditional 96-well plate to a 192-well design, significantly increasing throughput while maintaining operational convenience. It is suitable for high-volume cell culture and detection, improving experimental efficiency.

[0014] 2. In this utility model, a limiting part matching the automated equipment is set on the outer periphery of the substrate, and a fixing device including a hanging plate, a hanging rod, a telescopic rod and a return spring is installed on both sides. When in use, the hanging plate can be fixed in a suitable position, and the telescopic rod can be extended by pulling the hanging rod so that the hanging rod is inserted into the hanging plate. Then, the hanging rod is stably locked under the action of the return spring, realizing the rapid adaptation and stable installation of the substrate with different models of automated equipment, improving equipment compatibility and ease of operation. Attached Figure Description

[0015] Figure 1 This is a three-dimensional structural diagram of the present invention;

[0016] Figure 2 This utility model Figure 1 Enlarged structural diagram of part A in the middle;

[0017] Figure 3 This is a three-dimensional structural diagram of the fixing device of this utility model;

[0018] Figure 4 This utility model Figure 3 Enlarged structural diagram of part B.

[0019] Reference numerals: 1. Base plate; 2. Cell well; 3. Culture medium well; 4. Communicating groove; 5. Fixing device; 501. Connecting plate; 502. Mounting block; 503. Protrusion; 504. Movable spring; 505. Connecting rod; 506. Telescopic rod; 507. Hanging rod; 508. Pull-back spring; 509. Hanging plate. Detailed Implementation

[0020] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs; the terminology used herein in the specification of the application is for the purpose of describing particular embodiments only and is not intended to be limiting of the application; the terms "comprising" and "having," and any variations thereof, in the specification, claims, and foregoing drawings of this application are intended to cover non-exclusive inclusion. The terms "first," "second," etc., in the specification, claims, or foregoing drawings of this application are used to distinguish different objects, not to describe a particular order.

[0021] In this document, the term "embodiment" means that a particular feature, structure, or characteristic described in connection with an embodiment may be included in at least one embodiment of this application. The appearance of this phrase in various places throughout the specification does not necessarily refer to the same embodiment, nor is it a separate or alternative embodiment mutually exclusive with other embodiments. It will be explicitly and implicitly understood by those skilled in the art that the embodiments described herein can be combined with other embodiments.

[0022] This utility model provides a well plate for 3D cell culture, such as... Figures 1-4 As shown, it includes: a substrate 1, in which cell wells 2 and culture medium wells 3 are formed, and a connecting groove 4 is formed between the cell wells 2 and the culture medium wells 3. The connecting groove 4 is used to introduce buffer solution between the cell wells 2 and the culture medium wells 3 to reduce edge effect. A limiting part for cooperating with automated equipment is provided on the outer periphery of the substrate 1, and a fixing device 5 is installed on both sides of the substrate 1.

[0023] In this embodiment, the connecting groove 4 connects the cell well 2 and the culture medium well 3. By introducing buffer solution, a stable liquid bridge is formed, which effectively alleviates the mechanical disturbance during liquid exchange between the two wells, ensures a stable environment for cells and culture medium in the cell well 2, reduces shear force during operation, and thus significantly improves the survival rate and uniformity of cell culture.

[0024] In a further preferred embodiment of this invention, the volume of the culture medium well 3 is larger than the volume of the cell well 2, so as to provide more sufficient storage space for the culture medium.

[0025] In a further preferred embodiment of this utility model, the fixing device 5 includes a connecting plate 501, and mounting blocks 502 are symmetrically installed on the outside of the connecting plate 501. The mounting blocks 502 are fixedly connected to the base plate 1 by bolts. The connecting plate 501 has a protrusion 503 inside, and a movable spring 504 is provided on the outside of the protrusion 503. Two connecting rods 505 are fixedly connected to the outside of the movable spring 504. One end of the connecting rod 505 is fixedly connected to a telescopic rod 506. The telescopic ends of the two telescopic rods 506 are fixedly connected to the same hanging rod 507. A pull-back spring 508 is provided on the outside of the telescopic rod 506. One end of the pull-back spring 508 is fixedly connected to the outside of the connecting rod 505, and the other end of the pull-back spring 508 is fixedly connected to the outside of the hanging rod 507. A hanging plate 509 for fixing the hanging rod 507 is clamped on the outside of the hanging rod 507.

[0026] In this embodiment, the protrusion 503 serves as the load-bearing structure for the movable spring 504, effectively guiding the compression and rebound of the spring. This ensures that the connecting rod 505 maintains stable elastic force during extension and retraction, improving the response speed and durability of the telescopic mechanism and preventing loosening or jamming caused by structural loosening. The pull-back spring 508 covers the outside of the telescopic rod 506, and with the help of the elastic pull-back force of the spring, the telescopic rod 506 quickly resets after release, ensuring that the hanging rod 507 can automatically return to the preset position. This improves the reliability and ease of operation of the fixing process and prevents the telescopic rod 506 from remaining in a non-working position due to external interference.

[0027] In summary, by setting cell wells 2 and culture medium wells 3 as a paired structure and opening a connecting groove 4 between the two wells, buffer solution can be introduced between cell wells 2 and culture medium wells 3, so as to achieve nutrient transfer while avoiding direct disturbance of cells. When changing the culture medium, only the culture medium wells 3 need to be operated. The cells and fixative gel inside cell wells 2 remain stable, thereby significantly reducing the risk of cell loss and improving the reproducibility and reliability of the experiment. In this invention, a flow channel is formed between the outer periphery and the middle gap of the substrate 1 and filled with PBS buffer solution, so that the peripheral wells and the central wells are in the same culture environment. A limiting part matching the automated equipment is set on the outer periphery of the substrate 1, and a fixing device 5 including a hanging plate 509, a hanging rod 507, a telescopic rod 506 and a pull spring 508 is installed on both sides. In use, the hanging plate 509 can be fixed in a suitable position, and the telescopic rod 506 can be extended by pulling the hanging rod 507, so that the hanging rod 507 can be inserted into the hanging plate 509.

[0028] It is worth noting that the circuits, electronic components, and modules involved in this utility model are all existing technologies, which can be fully implemented by those skilled in the art, and need not be elaborated upon. The content protected by this utility model does not involve any improvement to the software and methods.

[0029] It should be understood that the disclosed apparatus can be implemented in other ways, given the several embodiments provided in this application. For example, the apparatus embodiments described above are merely illustrative; the division of units described above is only a logical functional division, and in actual implementation, there may be other division methods. For example, multiple units or components may be combined or integrated into another system, or some features may be ignored or not executed. Furthermore, the coupling or communication connections shown or discussed may be through some interfaces; the indirect coupling or communication connections between devices or units may be telecommunications or other forms.

[0030] The above embodiments are only used to illustrate the technical solutions of this utility model, and are not intended to limit the scope of protection of this utility model. Obviously, the described embodiments are only some embodiments of this utility model, not all embodiments. Based on these embodiments, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of this utility model. Although this utility model has been described in detail with reference to the above embodiments, those skilled in the art can still combine, add, delete, or otherwise adjust the features of the various embodiments of this utility model according to the circumstances without conflict or creative effort, thereby obtaining different technical solutions that do not fundamentally depart from the concept of this utility model. These technical solutions are also within the scope of protection of this utility model.

Claims

1. A well plate for 3D cell culture, characterized in that, include: The substrate (1) has cell pores (2) and culture medium pores (3) in it. A connecting groove (4) is provided between the cell pores (2) and the culture medium pores (3). The connecting groove (4) is used to introduce buffer solution between the cell pores (2) and the culture medium pores (3) to reduce edge effect. A limiting part for cooperating with automated equipment is provided on the outer periphery of the substrate (1). Fixing devices (5) are installed on both sides of the substrate (1).

2. A well plate for 3D cell culture according to claim 1, wherein, The volume of the culture medium well (3) is larger than that of the cell well (2) to provide more space for culture medium storage.

3. A well plate for 3D cell culture according to claim 1, wherein, The fixing device (5) includes a connecting plate (501), and mounting blocks (502) are symmetrically installed on the outside of the connecting plate (501). The mounting blocks (502) are fixedly connected to the base plate (1) by bolts. The connecting plate (501) has a protrusion (503) inside, and a movable spring (504) is provided on the outside of the protrusion (503).

4. A well plate for 3D cell culture according to claim 3, wherein, The movable spring (504) is externally fixedly connected to two connecting rods (505). One end of the connecting rod (505) is fixedly connected to a telescopic rod (506). The telescopic ends of the two telescopic rods (506) are fixedly connected to the same hanging rod (507). The telescopic rod (506) is covered with a pull-back spring (508).

5. A well plate for 3D cell culture according to claim 4, wherein, One end of the pull-back spring (508) is fixedly connected to the outside of the connecting rod (505), and the other end of the pull-back spring (508) is fixedly connected to the outside of the hanging rod (507).

6. A well plate for 3D cell culture as described in claim 4, characterized in that, The hanging rod (507) is fitted with a hanging plate (509) for fixing the hanging rod (507).

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