WO2021248637A1

WO2021248637A1 - Early embryo simulated fallopian tube environment in-vitro culture chip capable of breaking through growth retardation

Info

Publication number: WO2021248637A1
Application number: PCT/CN2020/103146
Authority: WO
Inventors: 黄海波; 顾鸣伟; 戴辞海; 朱易辰; 陈立国; 刘吉柱; 王阳俊
Original assignee: 苏州大学
Priority date: 2020-06-09
Filing date: 2020-07-21
Publication date: 2021-12-16
Also published as: CN111607516A; CN111607516B

Abstract

An early embryo simulated fallopian tube environment in-vitro culture chip capable of breaking through growth retardation, comprising a culture medium driving part and a culture chip body. The culture medium driving part comprises a hose track plate, a peristaltic pump, a hose, and two hose connectors; the peristaltic pump is mounted on the hose track plate; the hose is partially arranged in the hose track plate; two free ends of the hose are respectively connected to the two hose connectors. The culture chip body comprises a substrate, a positioning plate, a pipeline plate, a sealing plate, a cover plate, a fallopian tube epithelial cell culture cylinder, and a cumulus cell culture cylinder; the substrate, the positioning plate, the pipeline plate, and the sealing plate are tightly connected from bottom to top; the cover plate is connected to the sealing plate in an openable manner; the fallopian tube epithelial cell culture cylinder and the cumulus cell culture cylinder are both mounted on the pipeline plate.

Description

Early embryonic oviduct environment in vitro culture chip for breaking through developmental block

Technical field

The invention relates to the technical field of in vitro culture, in particular to an early embryonic oviduct environment in vitro culture chip for breaking through developmental block.

Background technique

Oocytes contain important biological genetic information needed for the reproduction of life, and are the origin of life gestation. The research on oocytes has always occupied a very important position in life science and medical research. With the development of science and technology, scientists have begun to study the in vitro culture of early embryos, and found that the in vitro culture of early embryos is very prone to developmental block and the survival rate has not been high. The so-called developmental retardation means that the reconstructed egg cell embryo starts to divide after being activated and develops from the 4-cell stage to the 8-cell stage. This period happens to be the transitional period of maternal gene-embryonic gene. The maternal mRNA in the embryonic cell is exhausted, and its own genome has not been activated yet. At this stage, the early embryo is extremely fragile, very sensitive to the external environment, and easily stops developing.

The inability of the existing in vitro embryo culture technology to provide a similar environment as that in the fallopian tube is the cause of a large number of developmental blockages in the process of in vitro embryo culture. In order to help the oocytes to pass through the developmental block in the in vitro embryo culture process, it is generally considered to be more effective to improve the composition of the culture medium, the co-cultivation of trophoblast somatic cells, dynamic culture and multi-ovum co-cultivation. Studies on the co-culture of trophoblast somatic cells have shown that the co-culture of oviduct epithelial cells can reduce the endogenous ROS of mouse embryos during the maternal-embryonic gene transition period, promote the transition from maternal to zygotic, and overcome embryo development in vitro. Dynamic culture can simulate the physical stimulation of the oviduct wall and cilia when the embryo is transported in the fallopian tube, such as using the shear force of the fluid to stimulate the embryo, which has a positive effect on the successful development of the embryo. Compared with static culture, the survival rate of the embryo is And the training speed has been improved. The autocrine or paracrine factors produced by the embryo will affect the development of the embryo and the rate of blastocyst formation. The interaction between the fertilized egg and the fertilized egg is very important. The fertilized egg co-cultured with multiple eggs has better performance than the fertilized egg cultured in isolation. Developmental potential.

A lot of research has been carried out on the dynamic culture system and experimental results have been obtained that have obvious advantages over the static culture system. However, in practical applications, the static training system still occupies a dominant position. The complexity of operation, the backwardness of equipment, and the inconvenience of use hinder the application and promotion of related technologies of the dynamic training system. However, with the development trend of egg cell manipulation technology, the success rate of the static culture system has been difficult to meet the demand.

Therefore, in view of the above technical problems, it is necessary to provide a chip for in vitro culture of early embryonic oviduct environment for breaking through developmental block.

Summary of the invention

In view of the shortcomings of the prior art, the purpose of the present invention is to provide a chip for in vitro culture of early embryonic oviduct environment for breaking through developmental block.

In order to achieve the foregoing objective, the technical solution provided by an embodiment of the present invention is as follows:

An early embryonic fallopian tube environment in vitro culture chip for breaking through developmental block, including a culture medium driving part and a culture chip body. The culture driving part includes a hose track plate, a peristaltic pump, a hose and two hose interfaces , The peristaltic pump is installed on the hose track plate, the hose is partially arranged in the hose track plate, and two free ends of the hose are respectively connected with the two hose interfaces, The culture chip body includes a substrate, a positioning plate, a piping plate, a sealing plate, a cover plate, a fallopian tube epithelial cell culture rack, and a cumulus cell culturing rack. The substrate, positioning plate, piping plate, and sealing plate are compact from bottom to top. Connected, the cover plate and the sealing plate can be opened and closed, and the fallopian tube epithelial cell culture rack and the cumulus cell culture rack are both installed on the piping plate.

As a further improvement of the present invention, the sealing plate is provided with a culture medium inlet hole, a culture medium outlet hole, and a sealing hole, and the two hose ports are respectively tightly fitted in the culture medium inlet hole and the culture medium outlet hole. , The cover plate and the sealing hole are hermetically matched.

As a further improvement of the present invention, the piping plate is provided with a culture medium inlet pool, a culture medium outlet pool, an inlet main channel, an outlet main channel, a culture tank, a fallopian tube epithelial cell culture rack placement groove, and a cumulus cell culture rack placement The culture medium inlet pool and the culture medium outlet pool are respectively arranged corresponding to the culture medium inlet hole and the culture medium outlet hole, and the culture medium inlet pool and the culture medium outlet pool are respectively connected to the inlet main channel and the outlet The main channel is connected, the inlet main channel and the outlet main channel are both connected to the culture tank through a shunt channel, the fallopian tube epithelial cell culture rack placement groove is arranged on the inlet main channel, and the cumulus cell The culturing rack placement groove is arranged on the culturing tank.

As a further improvement of the present invention, two supports are provided on the sealing plate, the two supports are connected with the cover plate by a rotating shaft, and the cover plate is provided with a boss, and the boss is connected to the cover plate. The sealing hole is sealed and matched.

As a further improvement of the present invention, a positioning array is provided on the positioning plate, the positioning array includes a plurality of positioning units, the plurality of positioning units are arranged in an array, and the plurality of positioning units extend into the culture chamber In the pool.

As a further improvement of the present invention, each of the positioning units includes a plurality of fan lobes, and the plurality of fan lobes form an open cylindrical structure.

As a further improvement of the present invention, the fallopian tube epithelial cell culture rack includes a hollow first cylinder, at least one first through hole group is arranged on the axial surface of the first cylinder, and the first through hole group includes A plurality of first through holes, the plurality of first through holes are arranged in a circular array, a plurality of second through hole groups are provided on the peripheral surface of the first cylinder, and the second through hole groups include a plurality of The second through holes are arranged in a circular array.

As a further improvement of the present invention, the cumulus cell culture rack includes a cumulus cell culture membrane scaffold and a cumulus cell culture membrane, and the cumulus cell culture membrane scaffold is connected to the surface of the cumulus cell culture membrane.

As a further improvement of the present invention, the cumulus cell culture membrane support includes a second cylinder and two support plates connected to the second cylinder, and the second cylinder closely fits with the culture tank. .

As a further improvement of the present invention, the cover plate, the sealing plate, the pipe plate and the positioning plate are respectively provided with a first threaded hole, a second threaded hole, a third threaded hole, and a fourth threaded hole.

The beneficial effects of the present invention are:

The invention cleverly designs the in vitro culture chip, divides it into a culture medium driving part and a culture chip body, realizes a compact and reasonable layout of each structure, and integrates the three major factors of the construct cytotrophoblast 3D environment, dynamic culture, and multi-egg co-cultivation. On the same chip, there are fallopian tube epithelial cell culture racks and cumulus cell culture racks in the pipeline plate, which are used to construct the 3D environment of the somatic trophoblast. The peristaltic pump is used to drive the flow of the medium to provide dynamic culture of oocytes. Environment, multiple oocytes are placed in a positioning array to achieve the effect of multi-egg co-cultivation. Not only can it be closer to the real natural environment of oocyte development, but also the internal relationship between various factors can be studied to achieve a better Good culture effect, breaks through the early embryonic development block, has many advantages such as beautiful appearance, complete functions, high efficiency, increase the success rate of in vitro culture, and portability.

Description of the drawings

In order to explain the embodiments of the present invention or the technical solutions in the prior art more clearly, the following will briefly introduce the drawings that need to be used in the description of the embodiments or the prior art. Obviously, the drawings in the following description are only These are some embodiments described in the present invention. For those of ordinary skill in the art, other drawings can be obtained based on these drawings without creative work.

Figure 1 is a schematic structural diagram of a preferred embodiment of the present invention;

2 is a schematic diagram of the structure of the culture medium driving part of the preferred embodiment of the present invention;

3 is a schematic diagram of the structure of the culture chip body of the preferred embodiment of the present invention;

4 is a schematic diagram of the structure of the cover plate of the preferred embodiment of the present invention;

Figure 5 is a schematic diagram of the structure of the sealing plate of the preferred embodiment of the present invention;

Figure 6 is a schematic structural diagram of a piping plate according to a preferred embodiment of the present invention;

Fig. 7 is a schematic structural diagram of a positioning plate of a preferred embodiment of the present invention;

Fig. 8 is an enlarged schematic diagram of A in Fig. 7;

Figure 9 is a schematic structural diagram of a fallopian tube epithelial cell culture rack according to a preferred embodiment of the present invention;

Figure 10 is a schematic structural diagram of a cumulus cell culture rack according to a preferred embodiment of the present invention;

Figure 11 is a schematic structural diagram of a cumulus cell culture membrane according to a preferred embodiment of the present invention;

Figure 12 is a partial enlarged schematic diagram of the cumulus cell culture membrane of the preferred embodiment of the present invention;

In the figure: 1. Culture medium drive part, 11, hose track plate, 12, peristaltic pump, 13, hose, 14, hose interface, 2. Culture chip body, 21, substrate, 22, positioning plate, 221, Positioning unit, 222, fourth threaded hole, 2211, fan flap, 23, pipe plate, 231, culture medium inlet tank, 232, culture medium outlet tank, 233, inlet main channel, 234, outlet main channel, 235, culture chamber Pool, 236, Fallopian tube epithelial cell culture rack placement slot, 237, Cumulus cell culture rack placement slot, 238, shunt channel, 239, third threaded hole, 24, sealing plate, 241, medium inlet hole, 242, medium Outlet hole, 243, sealing hole, 244, support, 245, second threaded hole, 25, cover, 251, boss, 252, first threaded hole, 26, fallopian tube epithelial cell culture rack, 261, first cylinder Body, 262, first through hole, 263, second through hole, 27, cumulus cell culture rack, 271, cumulus cell culture membrane support, 272, cumulus cell culture membrane, 2711, second cylinder, 2712 Support plate, 2713, support block, 2721, third cylinder, 2722, first channel, 2723, second channel, 2724, square hole, 28, shaft.

detailed description

In order to enable those skilled in the art to better understand the technical solutions of the present invention, the following will clearly and completely describe the technical solutions in the embodiments of the present invention with reference to the accompanying drawings in the embodiments of the present invention. Obviously, the described The embodiments are only a part of the embodiments of the present invention, rather than all the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative work shall fall within the protection scope of the present invention.

As shown in Figure 1, Figure 2, and Figure 3, an early embryonic oviduct-like environment in vitro culture chip for breaking through developmental block is used for in vitro culture of oocytes, including medium driving part 1 and culture chip body 2. The culture medium driving part 1 includes a hose track plate 11, a peristaltic pump 12, a hose 13, and two hose ports 14. The peristaltic pump 12 is installed on the hose track plate 11, and the hose 13 is partially arranged on the hose track plate 11. Wherein, the two free ends of the hose 13 are respectively connected to the two hose ports 14. The culture chip body 2 includes a base plate 21, a positioning plate 22, a pipe plate 23, a sealing plate 24, a cover plate 25, and a fallopian tube epithelial cell culture rack 26 And the cumulus cell culture rack 27, the base plate 21, the positioning plate 22, the pipe plate 23, and the sealing plate 24 are tightly connected from bottom to top, the cover plate 25 and the sealing plate 24 can be opened and closed, the fallopian tube epithelial cell culture rack 26 and the egg The mound cell culture racks 27 are all installed on the pipe plate 23.

As shown in Figure 5, the sealing plate 24 of the present invention is preferably provided with a culture medium inlet hole 241, a culture medium outlet hole 242, and a sealing hole 243. The cover plate 25 and the sealing hole 243 are tightly matched with each other, and the two hose ports 14 are respectively closely matched. In the medium inlet hole 241 and the medium outlet hole 242. In order to prevent the culture medium from flowing out, the outer surface of the two hose connections 14, the inner surface of the culture medium inlet hole 241 and the inner surface of the culture medium outlet hole 242 can be coated with impermeable paint.

In order to facilitate the opening and closing connection between the cover plate 25 and the sealing plate 24, the present invention preferably provides two supports 244 on the sealing plate 24. The two supports 244 are connected to the cover plate 25 by a rotating shaft 28, and the cover plate 25 is provided with The boss 251, as shown in FIG. 4, the boss 251 and the sealing hole 243 are hermetically fitted.

It is further preferred that the sealing hole 243 is provided at the center of the sealing plate 24, the medium inlet hole 241 and the medium outlet hole 242 are respectively located on the left and right sides of the sealing hole 243, and the two supports 244 are both located on the rear side of the sealing hole 243.

As shown in FIG. 6, the piping plate 23 of the present invention is preferably provided with a culture medium inlet tank 231, a culture medium outlet tank 232, an inlet main channel 233, an outlet main channel 234, a culture tank 235, and a fallopian tube epithelial cell culture rack placement groove 236. And the cumulus cell culture rack placement slot 237, the medium inlet pool 231 and the medium outlet pool 232 are respectively arranged corresponding to the medium inlet hole 241 and the medium outlet hole 242, the medium inlet pool 231 and the medium outlet pool 232 respectively It is connected to the inlet main channel 233 and the outlet main channel 234. The inlet main channel 233 and the outlet main channel 234 are both connected to the culture tank 235 through the shunt channel 238. The fallopian tube epithelial cell culture rack placement groove 237 is provided on the inlet main channel 234 , The cumulus cell culture rack placement slot 237 is set on the culture tank 235.

It is further preferred that the culture tank 235 is located at the center of the duct plate 23.

It is further preferred that the medium inlet pool 231 is coaxial with the medium inlet hole 241, and the medium outlet pool 232 is coaxial with the medium outlet hole 242, which facilitates the flow of the medium.

In the present invention, it is preferable that the diameter of the culture tank 235 is 4 to 5 mm, the diameter of the medium inlet tank 231 is 2 to 3 mm, the diameter of the medium outlet tank 232 is 2 to 3 mm, and the width of the main inlet channel 233 is 1.0 to 1.2 mm. The length of the main inlet channel 233 is 9-10mm, the width of a single channel in the shunt channel 238 is 0.3-0.4mm, the connection between the main inlet channel 233 and one of the shunt channels 238, and the connection point between the main outlet channel 234 and the other shunt channel 238 The length of the room is 15mm.

As shown in FIG. 9, the fallopian tube epithelial cell culture rack 26 includes a hollow first cylinder 261. At least one first through hole group is provided on the axial surface of the first cylinder 261, and the first through hole group includes a plurality of first through holes. Through holes 262, each first through hole 262 extends through the two axial surfaces of the first cylinder 261 in the axial direction, a plurality of first through holes 262 are arranged in a circular array, and the peripheral surface of the first cylinder 261 is provided There are a plurality of second through hole groups, the plurality of second through hole groups are arranged at intervals along the axial direction, the second through hole group includes a plurality of second through holes 263, and the plurality of second through holes 263 are arranged in a circular array, The second through hole 263 communicates with the first through hole 262. In this embodiment, two first through hole groups are provided on the axial surface of the first cylinder 261, and the two first through hole groups extend in the radial direction.

As shown in FIG. 10, the cumulus cell culture frame 27 includes a cumulus cell culture membrane support 271 and a cumulus cell culture membrane 272, and the cumulus cell culture membrane support 271 is connected to the surface of the cumulus cell culture membrane 272.

As shown in Figures 11 and 12, it is further preferred that the cumulus cell culture membrane support 271 includes a second cylinder 2711, two support plates 2712 connected to the upper part of the second cylinder 2711, and a bottom part of the second cylinder 2711. The two connected support blocks 2713 and the second cylinder 2711 are closely matched with the culture tank 235 to compress the volume of the culture tank 235, save the cost of the culture medium, and prevent the outflow of the culture medium.

More preferably, the diameter of the cumulus cell culture membrane 272 is 3 to 4 mm and the thickness is 0.1 to 0.2 mm. It is further preferred that the cumulus cell culture membrane 272 includes a third cylinder 2721. The side of the third cylinder 2721 is provided with two rows of channel assemblies, and each row of channel assemblies includes a plurality of parallel first channels 2722 and a plurality of parallel second channels. 2723, the first channel 2722 and the second channel 2723 are arranged vertically, and the plurality of first channels 2722 and the plurality of second channels 2723 are arranged in a vertical array. The sizes of the first channel 2722 and the second channel 2723 are all 0.06×0.03mm, The distance between the two first passages 2722 in the same row and the distance between the two second passages 2723 are both 0.09 mm. The upper and lower surfaces of the third cylinder 2721 have a plurality of square holes 2724 penetrating through them. The intersection of the passage 2723 communicates with the square hole 2724, and the size of the square hole 2724 is 0.15×0.15 mm.

It is further preferred that the extended space of the sealed hole 243 can enclose the culture tank 235, the fallopian tube epithelial cell culture rack placing groove 236, and the cumulus cell culture rack placing groove 237.

As shown in Figures 7 and 8, it is preferred that the positioning plate 22 of the present invention is provided with a positioning array. The positioning array includes a plurality of positioning units 221, which are arranged in an array, and the plurality of positioning units 221 extend into the culture tank. 235 within.

It is further preferred that each positioning unit 221 includes a plurality of fan lobes 2211, and the plurality of fan lobes 2211 form an open cylindrical structure.

It is further preferred that the positioning array includes nine positioning units 221, and the nine positioning units 221 are arranged in a 3×3 array. It is further preferred that the positioning unit 221 in the middle of the nine positioning units 221 is located at the center of the positioning plate 22. It is further preferred that each positioning unit 221 includes eight fan lobes 2211. It is further preferred that the inner diameter of the positioning unit 221 is 200-240μm, which matches the tip of a 0.5-10μl pipette, the positioning unit 221 can accommodate 1-2 oocytes, and the outer diameter of the positioning unit 221 is 240-300μm. The gap between the petals 2211 is 50-70 μm, and the height of each fan petal 2211 is 200-250 μm.

In the present invention, it is preferable that a first threaded hole 252, a second threaded hole 245, a third threaded hole 239, and a fourth threaded hole 222 are provided on the cover plate 25, the sealing plate 24, the piping plate 23, and the positioning plate 22, respectively. The first threaded hole 252, the second threaded hole 245, the third threaded hole 239, and the fourth threaded hole 222 have the same size. The second threaded hole 245, the third threaded hole 239, and the fourth threaded hole 222 are coaxial. When the 25 is in the closed state, the first threaded hole 252, the second threaded hole 245, the third threaded hole 239, and the fourth threaded hole 222 are concentric, and bolts (not shown in the figure) can be used to connect the cover plate 25 and the sealing plate 24 tightly connected together.

In the present invention, the thickness of the substrate 21 is preferably 2 to 3 mm, the thickness of the positioning plate 22 is 2 to 3 mm, the thickness of the duct plate 23 is 5 to 6 mm, the thickness of the sealing plate 24 is 3 to 4 mm, the substrate 21 is made of glass material, and the positioning plate 22, The pipe plate 23 and the sealing plate 24 are made of polydimethylsiloxane material.

The working principle of the present invention is as follows:

The fallopian tube epithelial cell culture rack 26 is implanted with oviduct epithelial cells for culture, and the cumulus cell culture rack 27 is implanted with cumulus cells for culture. A small amount of medium liquid is dropped into the positioning unit 221, and the oocytes are taken and put into In the positioning unit 221, the fallopian tube epithelial cell culture rack 26 is placed in the fallopian tube epithelial cell culture rack placing groove 236, the cumulus cell culture rack 27 is placed in the cumulus cell culture rack placing groove 237, the cover plate 25 is closed, and the cover plate is closed. The boss 251 on the 25 is in a sealed fit with the sealing hole 243 on the sealing plate 24, and the liquid is prevented by screwing the bolts into the first threaded hole 252, the second threaded hole 245, the third threaded hole 239, and the fourth threaded hole 222. Out, fix a hose interface 14 in the medium outlet hole 242, and fill the medium liquid from the medium inlet hole 241. When the culture chip body 2 and the hose 13 are both filled with medium liquid, another A hose interface 14 is fixed in the culture medium inlet hole 241, and the peristaltic pump 12 is turned on to connect it to the mains power, so that the culture medium flows in the in vitro culture chip and has a dynamic culture effect on the oocytes.

For those skilled in the art, it is obvious that the present invention is not limited to the details of the foregoing exemplary embodiments, and the present invention can be implemented in other specific forms without departing from the spirit or basic characteristics of the present invention. Therefore, from any point of view, the embodiments should be regarded as exemplary and non-limiting. The scope of the present invention is defined by the appended claims rather than the foregoing description, and therefore it is intended to fall within the claims. All changes within the meaning and scope of the equivalent elements of are included in the present invention. Any reference signs in the claims should not be regarded as limiting the claims involved.

In addition, it should be understood that although this specification is described in accordance with the implementation manners, not each implementation manner only includes an independent technical solution. This narration in the specification is only for clarity, and those skilled in the art should regard the specification as a whole The technical solutions in the various embodiments can also be appropriately combined to form other implementations that can be understood by those skilled in the art.

Claims

An early embryonic oviduct environment in vitro culture chip for breaking through developmental block, which is characterized by comprising a culture medium driving part and a culture chip body. The culture driving part includes a hose track plate, a peristaltic pump, a hose and two A hose interface, the peristaltic pump is installed on the hose track plate, the hose part is arranged in the hose track plate, two free ends of the hose are connected to the two hoses respectively The culture chip body includes a substrate, a positioning plate, a pipe plate, a sealing plate, a cover plate, a fallopian tube epithelial cell culture rack and a cumulus cell culture rack. The substrate, positioning plate, pipe plate, and sealing plate are Closely connected upwards in sequence, the cover plate and the sealing plate can be opened and closed, the fallopian tube epithelial cell culture rack and the cumulus cell culture rack are both installed on the piping plate.
The in vitro culture chip for early embryonic fallopian tube environment for breaking through developmental block according to claim 1, wherein the sealing plate is provided with a culture medium inlet hole, a culture medium outlet hole, and a sealing hole. The hose interface is tightly fitted in the culture medium inlet hole and the culture medium outlet hole respectively, and the cover plate is sealed and matched with the sealing hole.
The in vitro culture chip for the early embryonic fallopian tube environment for breaking through the developmental block of claim 1, wherein the piping plate is provided with a culture medium inlet pool, a culture medium outlet pool, an inlet main channel, and an outlet main channel. , Culture storage tank, fallopian tube epithelial cell culture rack placement slot and cumulus cell culture rack placement slot, the medium inlet pool and the medium outlet pool are respectively arranged corresponding to the medium inlet hole and the medium outlet hole, so The culture medium inlet pool and the culture medium outlet pool are respectively connected to the inlet main channel and the outlet main channel. The inlet main channel and the outlet main channel are both connected to the culture tank through a shunt channel, and the fallopian tube epithelium is The cell culture rack placing groove is arranged on the main entrance channel, and the cumulus cell culture rack placing groove is arranged on the culture tank.
The in vitro culture chip for the early embryonic fallopian tube environment for breaking through the developmental block according to claim 2, wherein the sealing plate is provided with two supports, and the two supports pass through the cover plate. The rotating shaft is connected, the cover plate is provided with a boss, and the boss is in a sealing fit with the sealing hole.
The early embryonic fallopian tube environment in vitro culture chip for breaking through developmental block according to claim 1, wherein a positioning array is provided on the positioning plate, and the positioning array includes a plurality of positioning units, and The positioning units are arranged in an array, and the multiple positioning units extend into the culturing tank.
According to claim 5, the early embryonic fallopian tube environment in vitro culture chip for breaking through the developmental block, wherein each of the positioning units includes a plurality of fan lobes, and the plurality of fan lobes form an open cylindrical structure .
The in vitro culture chip for an early embryonic fallopian tube environment oriented to break through developmental block according to claim 1, wherein the fallopian tube epithelial cell culture rack comprises a hollow first cylinder, and an axial surface of the first cylinder Is provided with at least one first through hole group, the first through hole group includes a plurality of first through holes, the plurality of first through holes are arranged in a circular array, and the peripheral surface of the first cylinder is arranged There are a plurality of second through hole groups, the second through hole group includes a plurality of second through holes, and the plurality of second through holes are arranged in a circular array.
The early embryonic fallopian tube environment in vitro culture chip for breaking through developmental block according to claim 1, wherein the cumulus cell culture rack comprises a cumulus cell culture membrane scaffold and a cumulus cell culture membrane, and the cumulus cell culture membrane The cumulus cell culture membrane support is connected to the surface of the cumulus cell culture membrane.
The in vitro culture chip for an early embryonic fallopian tube environment for breaking through developmental block according to claim 8, wherein the cumulus cell culture membrane scaffold comprises a second cylinder connected to the second cylinder Two supporting plates, the second cylinder is tightly matched with the culture tank.
The in vitro culture chip for the early embryonic fallopian tube environment for breaking through the developmental block according to claim 2, wherein the cover plate, the sealing plate, the piping plate, and the positioning plate are respectively provided with a first threaded hole and a second threaded hole. Threaded hole, third threaded hole, fourth threaded hole.