Embryonic microwell culture dish

Abstract

The utility model discloses an embryo micropore culture dish which can replace liquid to adjust mechanical force by rolling embryo, the embryo micropore culture dish includes: culture dish body and culture medium import and culture medium export which are arranged on the wall outside of culture dish body, at least one culture hole is arranged on the inner surface of the bottom surface of culture dish body, the culture hole is non-through hole, valve body is arranged at the connection of culture medium import and culture medium export and culture dish body, and the valve body controls the communication of culture medium import and culture medium export and culture dish body respectively. The structure of the embryo micropore culture dish can realize the replacement of culture medium, uses hemispherical recess culture, simulates balanced mechanical force, forms vortex to push embryo to roll, and simulates the rolling scene in vivo.

Description

Technical Field

[0001] This utility model relates to the technical field of preimplantation embryo culture devices, and more specifically, to an embryo microporous culture dish, particularly to an embryo microporous culture dish with a liquid replacement mechanism to adjust the mechanical force for embryo rolling. Background Technology

[0002] Embryo culture dishes refer to the culture of early embryos obtained in an in vitro artificial environment to monitor their development or to allow for artificial intervention in a developmental process. They can be used to prepare, store, manipulate, or transfer human gametes or embryos for in vitro fertilization or other assisted reproductive procedures.

[0003] Clinically, the currently used embryo culture dish is a 3.5cm diameter flat-bottomed dish. 20-30 μL / well of culture medium (G1 or G2) is added to the top using a glass tube, and the culture drop is covered with culture medium to reduce liquid evaporation and prevent mixing between different culture drops. The obtained fertilized eggs are then placed in a G1 culture drop (1 early embryo / 1 culture drop / 1 culture well). When the embryo reaches the appropriate stage, the early embryo is transferred from the G1 culture drop to a G2 culture drop in another dish using an embryo transfer tube. Except during the transfer stage, the embryo remains quiescent in the culture dish.

[0004] In the embryo culture dishes currently in use, the contact between the early embryo and the bottom of the culture dish is only a small point. However, in the body, the early embryo gradually moves from the fimbriae to the uterus in the fallopian tube, and is in a state of motion. Moreover, all sides of the entire embryo have the opportunity to come into contact with the inner layer of the fallopian tube.

[0005] Numerous studies have shown that changes in mechanical force have a significant impact on the establishment of cell polarity in early embryonic development, thereby affecting important physiological processes such as cell proliferation and lineage differentiation during early embryonic development. Mechanical force is considered a key driver of morphogenesis, and the physical properties between fluid pressure and cell density in mammalian embryos are crucial factors. Mechanical force plays an important role in embryonic differentiation; undifferentiated cells can sense the mechanical forces of their surrounding environment and differentiate in different directions according to the different stimuli.

[0006] During the development of the blastocyst from a fertilized egg, the fluid in the blastocoel originates from the fluid surrounding the embryo. This pressurized fluid shapes the embryo by disrupting the connections between cells: observations of intercellular proteins demonstrate that these proteins contact each other to hold cells tightly together. With the presence of fluid, these adhesion proteins rupture as the cells are pushed apart, and cells with fewer adhesion proteins are more easily separated. Utility Model Content

[0007] To address the aforementioned problems, this invention provides an embryo microporous culture dish. This culture dish features liquid replacement and allows the embryo to roll and adjust mechanical force, enabling real-time replenishment of the culture medium, allowing more surfaces of the early embryo to contact the culture dish, and enabling the embryo to roll within the culture dish.

[0008] To achieve the above objectives, this utility model provides the following technical solution: An embryo microporous culture dish with liquid replacement to regulate the mechanical force of embryo rolling, the embryo microporous culture dish comprising: a culture dish body, and a culture medium inlet and a culture medium outlet disposed on the outer side of the wall of the culture dish body; At least one culture hole is provided on the inner bottom surface of the culture dish body, and the culture hole is a non-penetrating hole. A valve is provided at the connection between the culture medium inlet and the culture medium outlet and the culture dish body, and the valve controls the connection between the culture medium inlet and the culture medium outlet and the culture dish body.

[0009] In an alternative embodiment, the culture dish body has 8-14 culture wells.

[0010] In an alternative embodiment, the culture well is a hemispherical depression.

[0011] In an alternative embodiment, the diameter of the culture well is 110-150 μm.

[0012] In an alternative embodiment, the culture medium added to each culture well is 3.49 × 10⁻⁶. -4 - 8.84 × 10 -4 μl.

[0013] In an alternative embodiment, the outer edge of the culture well has a smooth transition zone with the inner surface of the bottom of the culture dish body.

[0014] In an alternative embodiment, both the culture medium inlet and the culture medium outlet are L-shaped tubes, with the opening of the culture medium inlet facing upwards and the opening of the culture medium outlet facing downwards.

[0015] In an alternative embodiment, two through holes are horizontally formed on the wall of the culture dish body, and the culture medium inlet and culture medium outlet are respectively connected to the interior of the culture dish body through the through holes.

[0016] In an alternative embodiment, the valve body is mounted within a through hole.

[0017] In an alternative embodiment, the valve body is a one-way valve.

[0018] The beneficial effects of this utility model are as follows: This invention provides a microporous embryo culture dish with liquid replacement and the ability to regulate mechanical forces by rolling the embryo within the dish. This structure allows for real-time or on-demand replacement of the culture medium without moving the embryo, ensuring the stability of the culture medium's nutrient composition and pH value. It also ensures real-time liquid replenishment and allows more surface area of ​​the early embryo to contact the culture dish. Furthermore, the hemispherical concave culture method simulates balanced mechanical forces; the liquid flow creates eddies that propel the embryo to roll, mimicking the rolling motion within the embryo's body. Attached Figure Description

[0019] Figure 1 This is a schematic diagram of the structure of the embryo microporous culture dish of this utility model; Figure 2 This is a schematic cross-sectional view of the microporous embryo culture dish of this invention. Figure 3 This is a schematic diagram of the liquid culture medium flow in the embryo microporous culture dish of this invention. Figure 4 This is a diagram of the third embodiment of the embryo microporous culture dish of this utility model.

[0020] Figure labels: 1-Cultural dish body; 2-Cultural medium inlet; 21-Jet outlet; 3-Cultural medium outlet; 4-Cultural well; 5-Valve body; 6-Embryo; 7-Cultural medium layer; 8-Oil film layer. Detailed Implementation

[0021] To make the objectives, technical solutions, and advantages of this application clearer, the technical solutions of the present utility model will be clearly and completely described below with reference to the accompanying drawings of the embodiments of the present utility model. Obviously, the described embodiments are only some embodiments of the present utility model, and not all embodiments. Based on the embodiments of the present utility model, all other embodiments obtained by those of ordinary skill in the art without creative effort are within the scope of protection of the present utility model.

[0022] Embodiments of this utility model, examples of which are shown in the accompanying drawings, wherein the same or similar reference numerals denote the same or similar elements or elements having the same or similar functions throughout. The terms "first," "second," "third," etc., in the specification, claims, and drawings of this utility model (if present, are used to distinguish similar objects and are not necessarily used to describe a specific order or sequence. It should be understood that such described objects can be interchanged where appropriate. Furthermore, the terms "comprising" and "having," and any variations thereof, are intended to cover non-exclusive inclusion. Directional terms mentioned in this utility model, such as: up, down, left, right, front, back, inside, outside, side, etc., are only for the purpose of referring to the drawings. The embodiments described below with reference to the accompanying drawings are exemplary and are only used to explain this utility model, and should not be construed as limiting this utility model. Furthermore, the repetition of reference numerals and / or reference letters in different examples of this utility model is for simplification and clarity purposes and does not in itself indicate a relationship between the various embodiments and / or arrangements discussed. In addition, this utility model provides examples of various specific processes and materials, but those skilled in the art will recognize the application of other processes and / or the use of other materials.

[0023] like Figures 1-4 As shown, this utility model discloses an embryo microporous culture dish with liquid replacement to adjust the mechanical force of embryo rolling. The embryo microporous culture dish includes: a culture dish body 1, and a culture medium inlet 2 and a culture medium outlet 3 disposed on the outer side of the wall of the culture dish body 1.

[0024] As the embryo develops in its early stages, the nutrients in the culture medium are gradually consumed. Components such as pH, ion concentration, glucose, and sodium pyruvate are no longer optimal for early embryo development. Therefore, a culture medium inlet 2 and a culture medium outlet 3 are provided on the outer wall of the culture dish body 1 to allow the liquid to be updated in real time.

[0025] Because of the presence of a culture medium inlet 2 and a culture medium outlet 3, the liquid culture medium layer 7 can be replaced in real time or when needed without moving the embryo 6, and the stability of the culture medium's nutrients, pH value, etc. can be guaranteed.

[0026] Meanwhile, if the culture medium needs to be changed in the embryo culture dish, the embryos need to be transferred, which carries the risk of embryo loss or confusion. The culture medium inlet 2 and outlet 3 allow for culture medium changes without moving the embryos 6, thus avoiding this problem.

[0027] The petri dish body 1 is circular with a diameter of 3.5 cm. The petri dish body 1 has a circular bottom surface and walls extending upward from the circular bottom surface.

[0028] At least one culture well 4 is provided on the inner surface of the bottom of the culture dish body 1. The culture well 4 is a non-penetrating well. The embryo 6 is placed in the culture well 4.

[0029] One or more culture wells 4 can be provided on the inner surface of the bottom of the petri dish body 1. Optionally, the number of culture wells 4 is 8-14. Preferably, the petri dish body 1 is provided with 10 culture wells 4. The 10 culture wells 4 are evenly distributed on the bottom surface.

[0030] The projection shape of the culture well 4 onto the inner bottom surface of the culture dish body 1 can be of various shapes, such as a circle, a rectangle, or other shapes. Preferably, the projection shape of the culture well 4 is circular, and the culture well 4 is a hemispherical recess.

[0031] Unlike flat-bottomed embryo culture dishes, where only one point of the early embryo contacts the bottom, uneven mechanical forces can cause abnormalities in cell polarity and lineage differentiation. Using concave hemispherical culture wells 4 allows more of the early embryos 6 to contact the culture dish body 1.

[0032] Therefore, culturing embryo 6 in a hemispherical depression can simulate balanced mechanical forces. Simultaneously, the flow of the culture medium creates eddies that propel embryo 6 to roll, mimicking the rolling motion within the body.

[0033] Furthermore, depending on the size of the embryo 6, the diameter of each culture well 4 on the inner surface of the bottom surface ranges from 110 to 150 μm without affecting the vortex.

[0034] Preferably, the diameter of each culture well 4 on the inner surface of the bottom is 120 μm.

[0035] Furthermore, based on the commonly used clinical culture medium volume and the diameter of each culture well 4, using the hemispherical volume formula V = (2 / 3)πr³, where r is the radius of the culture well 4, the volume of culture medium added to each culture well 4 is found to be in the range of 3.49 × 10⁻⁶. -4 -8.84×10 -4 μl.

[0036] Preferably, the culture medium added to each culture well 4 is 5 × 10⁴ cm⁻¹. -4 μl.

[0037] like Figure 4 As shown, preferably, the outer edge of each culture well 4 has a smooth transition zone with the inner surface of the bottom of the culture dish body 1.

[0038] Early embryos are stationary in existing embryo culture dishes, which differs from their physiological state of gradually moving forward in the fallopian tubes. This stationary state affects the forces acting on the early embryo, thereby influencing cell polarity and lineage differentiation, ultimately impacting the early embryo's developmental potential. Unlike existing embryo culture dish structures, the embryo culture dish provided by this invention utilizes the flow of liquid culture medium to create a vortex at culture well 4, causing the embryo 6 to roll accordingly.

[0039] A valve body 5 is provided at the connection between the culture medium inlet 2 and the culture medium outlet 3 and the culture dish body 1. The valve body 5 controls the connection between the culture medium inlet 2 and the culture medium outlet 3 and the culture dish body 1, respectively.

[0040] Specifically, to prevent backflow of liquid culture medium, valve body 5 uses a one-way valve.

[0041] Preferably, both the culture medium inlet 2 and the culture medium outlet 3 are L-shaped tubes, with the opening of the culture medium inlet 2 facing upwards and the opening of the culture medium outlet 3 facing downwards.

[0042] By setting up a culture medium inlet 2 and a culture medium outlet 3, combined with a hemispherical concave culture well 4, fluid dynamics are used to create vortices at each culture well 4, which propel the early embryos 6 to roll.

[0043] In one specific embodiment, two through holes are horizontally formed on the wall of the culture dish body 1, and the culture medium inlet 2 and the culture medium outlet 3 are respectively connected to the interior of the culture dish body 1 through the through holes. The culture medium inlet 2 extends into the interior of the culture dish body 1 through the wall of the culture dish body 1, forming a jet port 21.

[0044] Furthermore, the inlet end of the culture medium inlet 2 is an injection gun interface, and the culture medium outlet 3 is a negative pressure outlet.

[0045] In the petri dish body 1, the culture medium layer 7 covers the culture well 4, and the upper surface of the culture medium layer 7 is higher than the communication point between the culture medium inlet 2 and the culture medium outlet 3 and the petri dish body 1.

[0046] Specifically, the culture medium layer 7 is also covered with an oil film layer 8.

[0047] This invention relates to a microporous embryo culture dish with liquid replacement that regulates the mechanical force of embryo rolling. The culture medium can be replaced in real time or as needed without moving the embryo, and the stability of the nutrient composition and pH value of the culture medium is ensured. The use of a hemispherical concave shape to culture the embryo can simulate a balanced mechanical force. At the same time, the flow of liquid culture medium forms a vortex that propels the embryo to roll, simulating the rolling scenario in vivo.

[0048] The technical features of the above embodiments can be combined in any way. For the sake of brevity, not all possible combinations of the technical features in the above embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

Claims

1. A microporous embryo culture dish, characterized in that, The embryo microporous culture dish includes: a culture dish body (1), and a culture medium inlet (2) and a culture medium outlet (3) disposed on the outer side of the wall of the culture dish body (1). At least one culture hole (4) is provided on the bottom inner surface of the culture dish body (1). The culture hole (4) is a non-penetrating hole, and each culture hole (4) contains an embryo (6). A valve body (5) is provided at the connection between the culture medium inlet (2) and the culture medium outlet (3) and the culture dish body (1). The valve body (5) controls the connection between the culture medium inlet (2) and the culture medium outlet (3) and the culture dish body (1).

2. The embryo microporous culture dish according to claim 1, characterized in that, The culture dish body (1) has 8-14 culture wells (4).

3. The embryo microporous culture dish according to claim 1, characterized in that, The culture well (4) is a hemispherical depression.

4. The embryo microporous culture dish according to claim 3, characterized in that, The diameter of the culture well (4) is 110-150 μm.

5. The embryo microporous culture dish according to claim 3, characterized in that, The culture medium added to each culture well (4) is 3.49 × 10⁻⁶. -4 -8.84×10 -4 μl.

6. The embryo microporous culture dish according to claim 1, characterized in that, The outer edge of the culture well (4) has a smooth transition zone with the inner surface of the bottom of the culture dish body (1).

7. The embryo microporous culture dish according to claim 1, characterized in that, The culture medium inlet (2) and the culture medium outlet (3) are both L-shaped tubes, with the opening of the culture medium inlet (2) facing upwards and the opening of the culture medium outlet (3) facing downwards.

8. The embryo microporous culture dish according to claim 1, characterized in that, The culture dish body (1) has two horizontal through holes on its wall. The culture medium inlet (2) and culture medium outlet (3) are connected to the interior of the culture dish body (1) through the through holes.

9. The embryo microporous culture dish according to claim 8, characterized in that, The end of the culture medium inlet (2) that extends into the body of the culture dish (1) is a jet port (21).

10. The embryo microporous culture dish according to claim 1, characterized in that, The valve body (5) is a one-way valve.

Images