Micro-fluidic chip for exploring in-vitro XY type sperm fertilization preference

Abstract

The invention discloses a micro-fluidic chip for exploring in-vitro XY type sperm fertilization preference, and relates to the technical field of biomedical engineering.According to the technical scheme, the micro-fluidic chip is characterized by comprising a chip body, the chip body is provided with a central sample carrying hole, the central sample carrying hole is used for containing an oocyte, and the central sample carrying hole is communicated with the central sample carrying hole; the chip body is provided with four peripheral sample carrying holes, the four peripheral sample carrying holes are used for placing fluorescently-labeled sperm cells subjected to sexual control sorting, and communicated micro-channels are arranged between the central sample carrying hole and the four peripheral sample carrying holes. Observation is facilitated by improving the volume of a liquid carrying hole and optimizing the transparency of a material, meanwhile, by adjusting the diameter of a channel and the number of repeated units, the thickness of the micro-fluidic chip is reduced, micro-channel blockage is avoided, the number of cells in loaded liquid is guaranteed, and the detection accuracy of the micro-fluidic chip is improved. The preference of successful fertilization of X and Y type sperms under different conditions is observed in vitro through micro-fluidic, and a new thought is provided for a method for controlling the gender of a fetus.

Description

Technical Field

[0001] The present invention relates to the technical field of biomedical engineering, and more specifically, it relates to a microfluidic chip for exploring the fertilization preference of XY sperm in vitro. Background Art

[0002] Microfluidic chip technology is an interdisciplinary subject involving fields such as chemistry, fluid physics, and biomedical engineering. This technology is a system that manipulates and controls tiny fluids (from μL to pL) using microchannels (with diameters in the tens to hundreds of micrometers) fabricated by microfabrication technology, characterized by miniaturization and automation.

[0003] Infertility is a widespread problem, and approximately 10%-15% of couples worldwide are troubled by it. Assisted reproductive technology, abbreviated as ART (Assisted Reproductive Technology), refers to the technology that uses medical assistance to enable infertile couples to become pregnant, including two major categories: artificial insemination (AI) and in vitro fertilization-embryo transfer (IVF-ET) and its derivative technologies. The main purpose of assisted reproductive technology is to help infertile couples successfully become pregnant and give birth to healthy offspring. It solves the difficulty of conception caused by physiological factors through medical means and provides these couples with the opportunity to have children.

[0004] In assisted reproductive technology, the selection of high-quality sperm is crucial for increasing the probability of fertilization and conception. These techniques include selection based on surface charge, sperm apoptosis, sperm birefringence, the ability to bind to hyaluronic acid, and sperm morphology under ultra-high magnification. Traditional sperm sample processing methods, such as centrifugation and washing, can introduce mechanical damage and oxidative stress, affecting sperm quality. These operations include sperm centrifugation, density gradient centrifugation, and the influence of certain components in the separation fluid. To reduce the impact of these operations, the screening of highly motile sperm should avoid centrifugation as much as possible.

[0005] Microfluidic chip technology can directly select high-quality sperm from raw semen by simulating the sperm selection mechanism in the female body, which can improve the efficiency of in vitro fertilization (IVF) and improve pregnancy outcomes, and has great clinical value. At the same time, microfluidic chip technology can effectively screen highly motile sperm by utilizing sperm chemotaxis and the characteristics of swimming along the interface.

[0006] However, currently, there is still a problem that it is difficult to detect and judge the preference of X-type and Y-type sperm during in vitro simulation of artificial insemination. Therefore, the present invention aims to provide a microfluidic chip for exploring the fertilization preference of XY-type sperm in vitro to solve the above problems. Summary of the Invention

[0007] The object of the present invention is to provide a microfluidic chip for exploring the fertilization preference of XY-type sperm in vitro. By improving the volume of the liquid loading holes, optimizing the transparency of the material for easy observation, and at the same time adjusting the channel diameter and the number of repeating units, the thickness of the microfluidic chip is greatly reduced, the microchannel blockage is reduced, the number of cells in the loaded liquid is guaranteed, and the preference of X- and Y-type sperm for successful fertilization under different conditions can be observed in the microfluidic body, which helps to provide new ideas for the method of fetal sex control.

[0008] The above technical object of the present invention is achieved through the following technical solutions: A microfluidic chip for exploring the fertilization preference of XY-type sperm in vitro, including a chip body, on which there is a central sample loading hole for placing oocytes, and four peripheral sample loading holes on the chip body for placing fluorescence-labeled sperm cells sorted by sex control. Microchannels are provided between the central sample loading hole and the four peripheral sample loading holes, and the microchannels are used to connect the central sample loading hole and the peripheral sample loading holes.

[0009] The present invention is further provided that: The chip body is driven by surface tension, and the surface tension of the fluid is used to attract sperm of different genders in the peripheral sample loading holes to migrate through the microchannels towards the oocytes in the central sample loading hole.

[0010] The present invention is further provided that: The diameter of the peripheral sample loading hole is 6 mm and the height is 3.5 mm.

[0011] The present invention is further provided that: The diameter of the microchannel is 0.151 mm and the height is 0.2 mm.

[0012] The present invention is further provided that: The chip body can always control the operating temperature at a constant 37 °C.

[0013] The present invention is further provided that: The microfluidic chip is made of PDMS material and has optical transparency, biocompatibility, and chemical inertness.

[0014] In summary, the present invention has the following beneficial effects:

[0015] 1. The microfluidic chip of the present invention can precisely control the migration path and speed of sperm, providing a stable in vitro simulation environment for studying the interaction between sperm and eggs. At the same time, through fluorescence labeling technology, X-type and Y-type sperm can be clearly distinguished, facilitating the observation and statistical analysis of sperm migration and fertilization preferences, and improving the accuracy and repeatability of the experiment.

[0016] 2. The microfluidic chip of the present invention can simulate different environmental factors, such as temperature, acidity and alkalinity, and small molecule compounds, to study the effects of these factors on sperm fertilization preferences, providing a new research tool for revealing the principles of fetal sex control in different species. At the same time, by optimizing the microchannel design and material selection, the risk of microchannel blockage is reduced, ensuring the number of cells in the loaded liquid, thereby improving the efficiency of the fertilization experiment.

[0017] 3. The microfluidic chip of the present invention is designed with optimized carrier liquid hole volume and transparency, facilitating fluorescence observation and subsequent data analysis, making the experimental results more intuitive and accurate. At the same time, the microfluidic chip can precisely control the number and swimming speed of sperm, facilitating the qualitative and quantitative statistics of fertilization results, providing reliable data support for the research.

[0018] 4. The present invention provides a new idea for the method of fetal sex control. Through in vitro simulation and experimental verification, it helps to develop new fertility technologies. At the same time, it provides an innovative platform for studying sperm biology, fertilization mechanism and sex determination mechanism, contributing to the development of the field of reproductive biology.

[0019] 5. The present invention can be applied to assisted reproductive technologies, providing a more efficient and precise fertility method for clinical practice to meet specific fertility needs. Compared with traditional in vitro fertilization technology, the microfluidic chip technology of the present invention can reduce costs, reduce the resources and time required for the experiment, and improve the success rate of the experiment. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 is a schematic external view of a microfluidic chip for exploring the fertilization preferences of XY-type sperm in vitro in an embodiment of the present invention;

[0021] Figure 2 is a schematic connection diagram of the central detection hole and the surrounding sample loading holes in an embodiment of the present invention.

[0022] In the figure: 1. Chip body; 2. Central detection hole; 3. Surrounding sample loading holes; 4. Microchannel. DETAILED DESCRIPTION OF THE EMBODIMENTS

[0023] The following is a further detailed description of the present invention with reference to the attached Figure 1-2 drawings.

[0024] Example: A microfluidic chip for exploring the fertilization preference of XY sperm in vitro, comprising a chip body 1. A central sample loading hole is fixedly arranged at the geometric center of the chip body 1 for placing oocytes. Four peripheral sample loading holes 3 are fixedly arranged around the central sample loading hole. The diameter of the peripheral sample loading hole 3 is 6 mm and the height is 3.5 mm. The four peripheral sample loading holes 3 are used for placing fluorescence-labeled sperm cells sorted by sex control. Microchannels are fixedly connected between the central sample loading hole and the four peripheral sample loading holes 3. The diameter of the microchannel is 0.151 mm and the height is 0.2 mm. The microchannel is used to connect the central sample loading hole and the peripheral sample loading hole 3. Among them, the chip body 1 can always control the operating temperature at a constant temperature of 37 °C. The chip body 1 is driven by surface tension, and the fluid surface tension is used to attract sperm of different genders in the peripheral sample loading hole 3 to migrate through the microchannel to the oocytes in the central sample loading hole.

[0025] Preferably, in this embodiment, the microfluidic chip is made of PDMS material. The whole model is made on a silicon wafer substrate by photolithography using the molding method, and then the PDMS prepolymer is poured into the model. After waiting for the prepolymer to cure, the PDMS is peeled off from the mold, and thus a microfluidic chip containing microchannels and sample loading holes is obtained, which has optical transparency, biocompatibility and chemical inertness. At the same time, sorting is carried out based on physical characteristics, and different male germ cells marked are distinguished by the pore diameter of the microchannel. The multi-channel parallel design improves efficiency. The main channel is connected to multiple branch channels, and the cell flow direction is dynamically allocated according to the sorting requirements. Each unit has four initial observation ports corresponding to different sorted cells.

[0026] Preferably, in this embodiment, by adding oocytes to the central sample loading hole and adding fluorescence-labeled X-type and Y-type sperm to the four peripheral sample loading holes 3 respectively, using the chemotaxis of sperm and the fluid surface tension, the sperm is attracted to migrate through the microchannel to the oocytes, and fluorescence detection is carried out at the pore diameter of the middle oocytes to judge the affinity ratio.

[0027] Preferably, in this embodiment, it also includes screening through different environmental factors in vitro, such as setting temperature differences, acid-base differences or adding different small molecule compounds, to study the fertilization preferences of X- and Y-type sperm under different environments. By simulating and studying the fertilization preferences of sperm for eggs in vitro, the design and operation method of this microfluidic chip helps to improve the experimental efficiency and can provide new ideas for the method of fetal sex control.

[0028] This specific embodiment is only an explanation of the present invention, and it is not a limitation of the present invention. Those skilled in the art can make modifications without creative contributions to this embodiment according to needs after reading this specification, but as long as it is within the scope of the claims of the present invention, it is protected by the patent law.

Claims

1. A microfluidic chip for exploring the fertilization preference of XY sperm in vitro, characterized in that: It includes a chip body (1), and a central sample loading hole is provided on the chip body (1). The central sample loading hole is used for placing oocytes. Four peripheral sample loading holes (3) are provided on the chip body (1), and the four peripheral sample loading holes (3) are used for placing fluorescence-labeled sperm cells that have been sex-sorted. Microchannels are provided between the central sample loading hole and the four peripheral sample loading holes (3), and the microchannels are used to connect the central sample loading hole and the peripheral sample loading holes (3).

2. The microfluidic chip for exploring the fertilization preference of XY sperm in vitro according to claim 1, characterized in that: The chip body (1) is driven by surface tension, and the surface tension of the fluid is used to attract sperm of different genders in the peripheral sample loading holes (3) to migrate through the microchannels towards the oocytes in the central sample loading hole.

3. A microfluidic chip for exploring the fertilization preference of XY sperm in vitro according to claim 1, characterized in that: The diameter of the peripheral sample loading hole (3) is 6 mm, and the height is 3.5 mm.

4. A microfluidic chip for exploring the fertilization preference of XY sperm in vitro according to claim 1, characterized in that: The diameter of the microchannel is 0.151 mm, and the height is 0.2 mm.

5. A microfluidic chip for exploring the fertilization preference of XY sperm in vitro according to claim 1, characterized in that: The chip body (1) can always control the operating temperature at a constant 37 °C.

6. The microfluidic chip for exploring the fertilization preference of XY sperm in vitro according to claim 1, characterized in that: The microfluidic chip is made of PDMS material and has optical transparency, biocompatibility, and chemical inertness.

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