Composite forming auxiliary reagent for assisting living tissue shaping and living tissue explant slice preparation method
By using composite molding auxiliary reagents to form hydrogel blocks of appropriate strength, the problem of uneven thickness of living tissue sections is solved, stable tissue cutting and cell integrity are achieved, and explant preparation of tissues of different hardness is suitable.
Patent Information
- Application Number
- CN202411441039.7
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-10-15
- Publication Date
- 2025-01-10
Smart Images
Figure CN119286763A_ABST
Abstract
Description
Technical Field
[0001] The invention belongs to the technical field of explant preparation, and in particular relates to a composite molding auxiliary reagent for assisting living tissue shaping and a living tissue explant slice preparation method. Background Art
[0002] In vitro models refer to physiological and pathological models constructed based on in vitro culture technology of various types of cells (such as primary human cells, cell lines, and functional cells derived from human pluripotent stem cells). They are tools that can simulate in vivo physiological conditions in vitro to study corresponding physiological, pathological, toxicological and other reactions. Preparing accurate experimental models is the key to understanding basic biology, disease development and treatment response, but generating a complex biological environment of an organ to study development or disease progression and treatment is also a challenging task. At present, common in vitro models mainly include ordinary 2D cultured cell models, 3D cultured cell models represented by organoids, and complex in vitro models represented by microfluidic organ chips. Among them, in vitro 2D cultured cells have always been an extremely important and widely used model. The cell source is mainly cells that can simulate the pathological manifestations of the corresponding disease and have the corresponding disease phenotype. However, 2D cultured cells are relatively weak in fitting the normal physiology of the human body and the microenvironment under disease conditions, and their response to external stimuli may be very different from the response of actual cells in the human body to the corresponding stimuli. Three-dimensional (3D) cultured cell models such as organoids and cell spheroids are three-dimensional constructs composed of single or multiple cell types. Compared with 2D cell models, 3D cultured cells are closer to the actual in vivo situation in terms of cell composition, cell distribution, cell behavior, and cell function. They can generate metabolic gradients, thereby producing good cell-to-cell and cell-ECM interactions. They can reproduce organ or pathological structures, tissue microenvironments, and even corresponding functions to a greater extent. However, due to the lack of stromal cells and immune cells, it is still a challenge for various 3D cultured cell models, including organoids, to simulate the tissue cell microenvironment in vitro.
[0003] The microenvironment in which human tissues survive and proliferate in the body has a matrix and complex multicellular components, which are difficult to fully summarize and reflect in the current in vitro cultured cells or organoid models. Therefore, in order to restore the in vivo tissue cell microenvironment as much as possible and meet the needs of subsequent testing, we have developed an in vitro model of human tissues that maintains cell viability and tissue structure, cell spatial network structure and distribution characteristics: tissue explants (Patient-driven tissue explants, PDEs).
[0004] In PDEs, the structure and microenvironment of the corresponding human tissues are completely preserved and maintain the spatial distribution characteristics consistent with the original tissues. They are cultured in vitro and subsequently processed and tested while maintaining vitality. For example, human tumor explants can survive for 12 days and maintain their original tumor, matrix and extracellular matrix structures, with functional multicellular stroma and fibrosis, without the need for complex tissue manipulation, digestion or artificial propagation of organoids. After cultivation, we can obtain drug sensitivity results by detecting the activity and functional changes of the corresponding target cells in the tissues after PDEs are treated with different drugs. This provides unprecedented possibilities for the study of disease mechanisms, preclinical research of new drugs, and precision medicine for various diseases.
[0005] PDEs are usually obtained by serial sectioning of patient-derived tissues. In the current conventional sample preparation method, the tissue is usually fixed directly on the sample holder and then immersed in the sample preparation buffer for PDE preparation. Fresh patient-derived living tissue is prepared as explant serial sections for in vitro culture and detection. Serial sections can ensure that each test group has consistent tissue cell layout and total cell number, and retain cell-to-cell interactions as much as possible. After the experiment, these PDEs can also be collected and paraffin-embedded or frozen sections more conveniently to meet the needs of subsequent pathological staining and immunohistochemical staining analysis.
[0006] However, different tissues have different degrees of hardness, and most tissues are relatively soft and difficult to shape. This makes it easy for these tissue blocks to deform significantly during the slicing process. Cells on the same plane may change position under the pressure of the cutting instrument, and the obtained PDEs may be uneven in thickness, which seriously affects their quality and thus affects subsequent operations and final test results. In addition, patient-derived tissues are valuable resources, and the methods of obtaining them include but are not limited to surgery and puncture. Among them, puncture specimens are usually small strips, and even surgically removed specimens are prone to tissue necrosis, calcification, or other conditions that may cause the entire tissue structure to loosen. After experiencing the inevitable bumps during transportation, these tissues may become smaller fragments, and even lose the opportunity to prepare PDEs and subsequent related tests by conventional methods. Summary of the invention
[0007] The present invention provides a composite molding auxiliary reagent for assisting the shaping of living tissue and a method for preparing living tissue explant slices, aiming to solve the technical problem in the prior art that uneven slice thickness is easily caused when preparing explant continuous slices due to the softness or loose structure of the living tissue.
[0008] In order to solve the above technical problems, the technical solution adopted by the present invention is:
[0009] In a first aspect, the present invention provides a composite molding auxiliary reagent for assisting the shaping of living tissue, comprising: 4-15 parts by mass of methacryloyl polyether F127 diacrylate F127DA, 2.5-5 parts by mass of GelMA gelatin aqueous solution, 4-6 parts by mass of gelatin, 2.5-5 parts by mass of AlgMA sodium alginate hydrogel, 0.20-0.30 parts by mass of LAP photoinitiator and 95-105 parts by volume of buffer solution.
[0010] In a preferred embodiment, the composite molding auxiliary reagent includes 4 parts by mass of methacrylated polyether F127 diacrylate F127DA, 2.5 parts by mass of GelMA gelatin aqueous solution, 5 parts by mass of gelatin, 2.5 parts by mass of AlgMA sodium alginate hydrogel, 0.25 parts by mass of LAP photoinitiator and 100 parts by volume of PBS buffer.
[0011] In a preferred embodiment, the composite molding auxiliary reagent includes 12 parts by mass of methacrylated polyether F127 diacrylate F127DA, 2.5 parts by mass of GelMA gelatin aqueous solution, 5 parts by mass of gelatin, 3.5 parts by mass of AlgMA sodium alginate hydrogel, 0.30 parts by mass of LAP photoinitiator and 105 parts by volume of PBS buffer.
[0012] Based on the above scheme, the mixed hydrogel system can meet the strength and toughness requirements of explant preparation.
[0013] In a second aspect, the present invention provides a method for preparing continuous sections of living tissue explants, comprising:
[0014] S1, dissolving 4-15 parts by mass of methacryloyl polyether F127 diacrylate F127DA, 2.5-5 parts by mass of GelMA gelatin aqueous solution, 4-6 parts by mass of gelatin, 2.5-5 parts by mass of AlgMA sodium alginate hydrogel and 0.20-0.30 parts by mass of LAP photoinitiator in 100 parts by volume of PBS buffer to obtain a composite molding auxiliary reagent;
[0015] S2, adding the composite molding auxiliary reagent into a transparent container, adding the tissue block to completely immerse it in the composite molding auxiliary reagent, and irradiating it with a UV light source for 30-60 seconds to photocured gel to obtain a tissue hydrogel block;
[0016] S3, placing the tissue hydrogel block on a slicing instrument for slicing to obtain living tissue explant slices;
[0017] Here, 1 part by mass: 1 part by volume = 1 g: 1 mL.
[0018] According to a preferred embodiment, in step S1, the composite molding auxiliary reagent includes 4 parts by mass of methacrylated polyether F127 diacrylate F127DA, 2.5 parts by mass of GelMA gelatin aqueous solution, 5 parts by mass of gelatin, 2.5 parts by mass of AlgMA sodium alginate hydrogel and 0.25 parts by mass of LAP photoinitiator dissolved in 100 parts by volume of PBS buffer to obtain the composite molding auxiliary reagent.
[0019] In a preferred embodiment, in step S2, after obtaining the tissue hydrogel block, a marking liquid is prepared to mark the tissue hydrogel block;
[0020] The labeling solution comprises 0.5-1 parts by mass of gelatin, 0.00004-0.0001 parts by mass of a water-soluble pigment solution and 10 parts by volume of a PBS buffer solution;
[0021] Here, 1 part by mass: 1 part by volume = 1 g: 1 mL.
[0022] Based on the above scheme, the use of the marking liquid can assist in determining the position when preparing and transferring tissue explant slices to a culture device.
[0023] In a preferred embodiment, the water-soluble pigment solution is phenol red.
[0024] Based on the above scheme, phenol red colors quickly, lasts for a long time, and has obvious color.
[0025] In a preferred embodiment, in step S2, a tissue hydrogel block is obtained by irradiating the gel with a UV light source for 30 seconds to photo-cure the gel.
[0026] Based on the above scheme, using UV light source for curing for too long will cause the tissue hydrogel block to be too hard and difficult to cut. Using UV light source for curing for too short will cause the tissue hydrogel block to be incompletely solidified, affecting cutting. When the UV light source curing time is 30s, the tissue hydrogel block has moderate hardness after solidification, which is conducive to cutting.
[0027] Preferably, in step S3, the wavelength of the UV light source is 365-405 nm.
[0028] In a preferred embodiment, the living tissue is soft tissue or fragmented tissue.
[0029] The beneficial effects of the present invention are:
[0030] 1. The present invention provides a composite forming auxiliary reagent for assisting the shaping of living tissue and a method for preparing living tissue explant slices. The composite forming auxiliary reagent can quickly gel under ultraviolet light conditions after the tissue block is immersed to form a hydrogel block with suitable strength and toughness, which can ensure that the entire structure has a certain hardness and toughness, so that the tissue block does not deform excessively under pressure during the cutting process, and can also avoid excessive strength causing additional damage or even death of cells in the tissue under squeezing, and can also avoid gel fragmentation caused by cutting, so as to obtain uniform thickness of living tissue explant slices. By changing the light curing time or changing the final concentration of the corresponding components, the strength of the hydrogel block can be adjusted, thereby meeting the tissue explant preparation requirements of tissue specimens with different hardness, and avoiding the instability of tissue explant preparation caused by the hardness difference between the peripheral gel and the central tissue specimen.
[0031] 2. The composite molding auxiliary reagent provided by the present invention also has excellent biocompatibility. The outer circle of the tissue explant prepared after cutting is wrapped with a circle of solidified hydrogel, which can be clamped and placed flat in a culture device to continue to be cultured for a period of time. The gel can stably absorb the components of the culture medium, assist in maintaining the shape structure of the explant and keep the surface of the explant moist, while not affecting the biological activity and characteristics of the culture. BRIEF DESCRIPTION OF THE DRAWINGS
[0032] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings required for use in the embodiments are briefly introduced below. It should be understood that the following drawings only show certain embodiments of the present invention and therefore should not be regarded as limiting the scope. For ordinary technicians in this field, other relevant drawings can be obtained based on these drawings without paying creative work.
[0033] Figure 1 This is a picture of a mouse liver biopsy tissue pretreatment sample successfully prepared in the present invention.
[0034] Figure 2 It is a schematic diagram of the pre-treated sample successfully transferring mouse liver living tissue to the stage of the slicer instrument in the present invention.
[0035] Figure 3 This is a slice diagram of a successfully prepared liver tissue explant.
[0036] Figure 4 This is a magnified image of mouse liver PDEs under a microscope.
[0037] Figure 5 This is an enlarged image of human gastric cancer PDEs under a microscope. DETAILED DESCRIPTION
[0038] The following will be combined with the accompanying drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not used to limit the present invention. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative work belong to the protection scope of the present invention.
[0039] Embodiment 1:
[0040] The present embodiment provides a composite molding auxiliary reagent for assisting the shaping of living tissue, including: 4-15 parts by mass of methacryloyl polyether F127 diacrylate F127DA, 2.5-5 parts by mass of GelMA gelatin aqueous solution, 4-6 parts by mass of gelatin, 2.5-5 parts by mass of AlgMA sodium alginate hydrogel, 0.20-0.30 parts by mass of LAP photoinitiator and 95-105 parts by volume of buffer solution.
[0041] The composite molding auxiliary reagent includes 4 parts by mass of methacrylated polyether F127 diacrylate F127DA, 2.5 parts by mass of GelMA gelatin aqueous solution, 5 parts by mass of gelatin, 2.5 parts by mass of AlgMA sodium alginate hydrogel, 0.25 parts by mass of LAP photoinitiator and 100 parts by volume of PBS buffer.
[0042] Alternatively, the composite molding auxiliary reagent includes 6 parts by mass of methacrylated polyether F127 diacrylate F127DA, 5 parts by mass of GelMA gelatin aqueous solution, 5 parts by mass of gelatin, 4 parts by mass of AlgMA sodium alginate hydrogel, 0.25 parts by mass of LAP photoinitiator and 100 parts by volume of PBS buffer.
[0043] Alternatively, the composite molding auxiliary reagent includes 8 parts by mass of methacrylated polyether F127 diacrylate F127DA, 3.5 parts by mass of GelMA gelatin aqueous solution, 4.5 parts by mass of gelatin, 3 parts by mass of AlgMA sodium alginate hydrogel, 0.30 parts by mass of LAP photoinitiator and 95 parts by volume of PBS buffer.
[0044] Alternatively, the composite molding auxiliary reagent includes 12 parts by mass of methacrylated polyether F127 diacrylate F127DA, 2.5 parts by mass of GelMA gelatin aqueous solution, 5 parts by mass of gelatin, 3.5 parts by mass of AlgMA sodium alginate hydrogel, 0.30 parts by mass of LAP photoinitiator and 105 parts by volume of PBS buffer.
[0045] In addition, the specific ratio can be appropriately adjusted according to the hardness of the tissue block to be processed.
[0046] Embodiment 2:
[0047] This embodiment provides a method for preparing continuous sections of living tissue explants, comprising:
[0048] S1, dissolving 4-15 parts by mass of methacryloyl polyether F127 diacrylate F127DA, 2.5-5 parts by mass of GelMA gelatin aqueous solution, 4-6 parts by mass of gelatin, 2.5-5 parts by mass of AlgMA sodium alginate hydrogel and 0.20-0.30 parts by mass of LAP photoinitiator in 100 parts by volume of PBS buffer to obtain a composite molding auxiliary reagent;
[0049] S2, adding the composite molding auxiliary reagent into a transparent container, adding the tissue block to completely immerse it in the composite molding auxiliary reagent, and irradiating it with a UV light source for 30-60 seconds to photocured gel to obtain a tissue hydrogel block;
[0050] Wherein, the wavelength of the UV light source is 365-405nm.
[0051] S3, placing the tissue hydrogel block on a slicing instrument for slicing to obtain living tissue explant slices;
[0052] Here, 1 part by mass: 1 part by volume = 1 g: 1 mL.
[0053] In step S1, the composite molding auxiliary reagent includes 8 parts by mass of methacrylated polyether F127 diacrylate F127DA, 3.5 parts by mass of GelMA gelatin aqueous solution, 5 parts by mass of gelatin, 3.5 parts by mass of AlgMA sodium alginate hydrogel and 0.25 parts by mass of LAP photoinitiator dissolved in 100 parts by volume of PBS buffer to obtain a composite molding auxiliary reagent.
[0054] In addition, in step S2, after obtaining the tissue hydrogel block, a labeling liquid is prepared to label the tissue hydrogel;
[0055] The labeling solution comprises 0.5-1 parts by mass of gelatin, 0.00004-0.0001 parts by mass of a water-soluble pigment solution and 10 parts by volume of a PBS buffer solution;
[0056] Here, 1 part by mass: 1 part by volume = 1 g: 1 mL.
[0057] Specifically, a space for dripping the marking liquid is reserved before the tissue hydrogel block solidifies, and a marking rod or other tool can be used to occupy the space before the tissue hydrogel solidifies. After the tissue hydrogel solidifies, the marking rod is pulled out, and then the marking liquid is injected into the reserved space in the obtained tissue hydrogel block, and a marked tissue gel block is obtained after solidification. This ensures that when the tissue clot is sliced later, each slice has a marking point.
[0058] Wherein, the water-soluble pigment solution is phenol red.
[0059] Specifically, the water-soluble pigment solution may be a water-soluble pigment solution with good biocompatibility and color development, and may be phenol red, lemon yellow, carmine, amaranth, sunset yellow or erythrosine, etc. A more preferred solution: the water-soluble pigment solution used in this embodiment is phenol red.
[0060] In addition, in step S2, a UV light source is used to irradiate for 30 seconds to photocurate the gel to obtain a tissue hydrogel block.
[0061] Specifically, the tissue hydrogel block is obtained by irradiating the gel with a UV light source for 30 seconds, 45 seconds or 60 seconds to photocurate the gel. In a more preferred solution, the tissue hydrogel block is obtained by irradiating the gel with a UV light source for 30 seconds to photocurate the gel.
[0062] Wherein, the living tissue is soft tissue or fragmented tissue.
[0063] The present invention will be further explained below in conjunction with specific experiments:
[0064] 1. Tissue sampling and cleaning
[0065] 1.1 Mouse tissue
[0066] Preparation: sterilize ophthalmic scissors and ophthalmic forceps in advance with high temperature and high pressure; turn on the ultraviolet light for 30 minutes in the clean bench; ice box; ice; microelectronic scale; 100mg / ml antibiotic (Primocin TM ,) sterile PBS buffer refrigerated at 4℃ for future use; 60mm sterile culture dishes and tissue preservation solution.
[0067] The specific steps for mouse tissue sampling and cleaning are as follows:
[0068] (1) Six-week-old C57 mice were killed by cervical dislocation and placed in a clean bench after being soaked in alcohol for 3-5 seconds;
[0069] (2) Use ophthalmic forceps to grasp the abdominal skin, and use ophthalmic scissors to cut the mouse's abdominal skin along the middle of the abdomen. Then cut the mouse's abdomen along the linea alba to expose the abdominal organs. Remove one lobe of the mouse's liver completely, wash it three times in PBS to remove as much excess blood as possible, and then store it in tissue preservation solution for later use.
[0070] 1.2 Human gastric cancer tissue
[0071] Human gastric cancer tissue was obtained from the lesion site of clinical gastric cancer patients by surgical sampling. The tissue was stored for 7 minutes after sampling and transported to the laboratory at 2-8°C for subsequent processing.
[0072] 2. Preparation of composite molding auxiliary reagents
[0073] Preparation: Methacrylated gelatin (GelMA), methacrylated polyether F127 diacrylate (F127DA), gelatin, sodium alginate hydrogel (AlgMA), photoinitiator LAP, 100 mg / ml antibiotic (Primocin TM ) sterile PBS buffer, 0.22 μm filter, biosafety cabinet with UV light turned on for 30 minutes, preheated water bath (37°C), 5 mL syringe, pipette, pipette tip, ice box and ice.
[0074] 2.1 The specific steps for preparing the auxiliary reagents for mouse liver tissue composite molding are as follows:
[0075] Weigh 0.4 g of methacryloyl polyether F127 diacrylate (F127DA), 0.25 g of gelatin hydrogel (GelMA), 0.5 g of gelatin, 0.25 g of sodium alginate hydrogel (AlgMA) and 0.025 g of photoinitiator LAP, then add 10 mL of PBS buffer to dissolve, mix the liquid after dissolution, and then filter and sterilize with a 0.22 μm filter.
[0076] 2.2 The specific steps for preparing the human gastric cancer tissue composite molding auxiliary reagent are as follows:
[0077] Weigh 0.6 g of methacryloyl polyether F127 diacrylate (F127DA), 0.5 g of gelatin hydrogel (GelMA), 0.5 g of gelatin, 0.4 g of sodium alginate hydrogel (AlgMA) and 0.025 g of photoinitiator LAP, then add 10 mL of PBS buffer, mix the liquid after dissolution, and then filter and sterilize with a 0.22 μm filter.
[0078] 3. Preparation of marking solution
[0079] (1) Weigh 0.5 g of gelatin and 6 μg of phenol red, then add 10 mL of PBS buffer and mix thoroughly;
[0080] (2) After the above materials are dissolved, mix the liquid and then filter it with a 0.22 μm filter for sterilization;
[0081] (3) Place the obtained labeled solution in a 37°C water bath for later use.
[0082] 4. Pretreatment of living tissue embedding
[0083] Preparation: Turn on the UV light in the biosafety cabinet for 30 minutes in advance; contain 100 mg / ml antibiotics (Primocin TM) sterile PBS buffer, the above-mentioned composite molding auxiliary reagents; a UV curing light source, tweezers and a pre-treated mold.
[0084] The specific steps of pretreatment for embedding living tissue are as follows:
[0085] (1) Add 0.5 mL of composite molding auxiliary reagent into the container, wait for it to solidify slightly and become viscous, place the tissue block on the gel, and then continue to add composite molding auxiliary reagent into the mold until the liquid surface submerges the tissue block;
[0086] (2) Irradiate the pre-treated mold containing the composite molding auxiliary reagent and tissue block with a UV curing light source for about 30 seconds.
[0087] (3) Add the marking solution dropwise into the container and then refrigerate the entire container at 2-8°C for 3-5 minutes.
[0088] (4) After all the reagents in the container have solidified, a shaped gel block with a certain strength is obtained.
[0089] 5. Preparation and identification of PDEs
[0090] Preparation: The ultra-clean workbench was pre-opened with UV for 30 minutes; a vibrating slicer for living tissue; curved forceps, a pipette, and a 100 mg / ml antibiotic (Primocin TM ,) sterile PBS buffer, pipette tips, super glue, pre-cooled freezing section machine, OCT embedding agent, HE staining reagent and adsorption slides.
[0091] The specific steps for preparing explant slices are as follows:
[0092] (1) Use tweezers to transfer the pre-treated embedded tissue block to the stage of the slicer and glue it firmly with super glue;
[0093] (2) Install the stage into the slicer slot of the slicer and inject PBS buffer into the slot until the buffer is soaked with tissue gel;
[0094] (3) starting the vibrating microtome for rapid slicing with a slice thickness of 250 μm;
[0095] (4) The explants obtained by sectioning were flushed off the blade with PBS buffer to observe the integrity of PDEs;
[0096] (5) After collection, the slices are stacked one by one, and then OCT embedding medium is added. The longitudinal sections of the PDEs are frozen and stained with HE to evaluate the thickness uniformity of the prepared PDEs.
[0097] 6. Test results
[0098] The results of PDEs in mouse liver tissue are as follows Figure 1-Figure 4 The PDEs in the mouse liver tissue obtained were uniform, and the slice thickness was about 255 μm.
[0099] The results of human gastric cancer PDEs are as follows Figure 5 As shown, the results of HE staining on the left and Ki67 immunohistochemical staining on the right demonstrated that the cell proliferation activity in human gastric cancer PDEs was good and the spatial distribution characteristics of the cells were preserved.
[0100] The present invention is not limited to the above-mentioned optional implementation modes. Anyone can derive other various forms of products under the inspiration of the present invention. However, no matter what changes are made in the shape or structure, all technical solutions that fall within the scope defined by the claims of the present invention fall within the protection scope of the present invention.
Claims
1. A composite molding auxiliary agent for assisting the shaping of living tissue, characterized in that: include: 4-15 parts by weight of methacrylated polyether F127 diacrylate F127DA, 2.5-5 parts by weight of GelMA gelatin aqueous solution, 4-6 parts by weight of gelatin, 2.5-5 parts by weight of AlgMA sodium alginate hydrogel, 0.20-0.30 parts by weight of LAP photoinitiator and 95-105 parts by volume of buffer.
2. A composite molding auxiliary agent for assisting the shaping of living tissue according to claim 1, characterized in that: The method comprises 4 parts by mass of methacrylated polyether F127 diacrylate F127DA, 2.5 parts by mass of GelMA gelatin aqueous solution, 5 parts by mass of gelatin, 2.5 parts by mass of AlgMA sodium alginate hydrogel, 0.25 parts by mass of LAP photoinitiator and 100 parts by volume of PBS buffer.
3. A composite molding auxiliary agent for assisting the shaping of living tissue according to claim 1, characterized in that: The method comprises 12 parts by weight of methacrylated polyether F127 diacrylate F127DA, 2.5 parts by weight of GelMA gelatin aqueous solution, 5 parts by weight of gelatin, 3.5 parts by weight of AlgMA sodium alginate hydrogel, 0.30 parts by weight of LAP photoinitiator and 105 parts by volume of PBS buffer.
4. A method for preparing continuous sections of living tissue explants, characterized in that: include: S1, dissolving 4-15 parts by mass of methacryloyl polyether F127 diacrylate F127DA, 2.5-5 parts by mass of GelMA gelatin aqueous solution, 4-6 parts by mass of gelatin, 2.5-5 parts by mass of AlgMA sodium alginate hydrogel and 0.20-0.30 parts by mass of LAP photoinitiator in 100 parts by volume of PBS buffer to obtain a composite molding auxiliary reagent; S2, adding the composite molding auxiliary reagent into a transparent container, adding the tissue block to completely immerse it in the composite molding auxiliary reagent, and irradiating it with a UV light source for 30-60 seconds to photocured gel to obtain a tissue hydrogel block; S3, placing the tissue hydrogel block on a slicing instrument for slicing to obtain living tissue explant slices; Here, 1 part by mass: 1 part by volume = 1 g: 1 mL.
5. A method for preparing continuous sections of living tissue explants according to claim 4, characterized in that: In step S1, the composite molding auxiliary reagent includes 4 parts by mass of methacrylated polyether F127 diacrylate F127DA, 2.5 parts by mass of GelMA gelatin aqueous solution, 5 parts by mass of gelatin, 2.5 parts by mass of AlgMA sodium alginate hydrogel and 0.25 parts by mass of LAP photoinitiator dissolved in 100 parts by volume of PBS buffer to obtain a composite molding auxiliary reagent.
6. A method for preparing continuous sections of living tissue explants according to claim 4, characterized in that: In step S2, after obtaining the tissue hydrogel block, a marking liquid is prepared to mark the inner or outer wall of the tissue hydrogel block, wherein the marking liquid; The labeling solution comprises 0.5-1 parts by mass of gelatin, 0.00004-0.0001 parts by mass of a water-soluble pigment solution and 10 parts by volume of a PBS buffer solution; Here, 1 part by mass: 1 part by volume = 1 g: 1 mL.
7. A method for preparing continuous sections of living tissue explants according to claim 4, characterized in that: The water-soluble pigment solution is phenol red.
8. A method for preparing continuous sections of living tissue explants according to claim 4, characterized in that: In step S2, a tissue hydrogel block is obtained by irradiating with a UV light source for 30 seconds to photocurate the gel.
9. A method for preparing continuous sections of living tissue explants according to claim 4, characterized in that: In step S3, the wavelength of the UV light source is 365-405 nm.
10. A method for preparing continuous sections of living tissue explants according to claim 1 or 4, characterized in that: The living tissue is soft tissue or fragmented tissue.