1533 results about "Cryopreservation" patented technology

The invention discloses a porcine reproductive and respiratory syndrome virus N protein antigen epitope peptide and a monoclonal antibody thereof. The amino acid sequence of the antigen epitope peptide is as shown in SEQ ID No. 11; the heavy chain variable region of the monoclonal antibody comprises CDR1-3 as shown in SEQ ID No.1 to SEQ ID No.3; the light chain variable region comprises CDR1-3 as shown in SEQ ID No. 4 to 6; the monoclonal antibody reacts with a variety of PRRSV strains such as classical PRRSV, HP-PRRSV, NADC30PRRSV-like, NADC34PRRSV-like, European PRRSV and the like, and the conservative property is good; the antibody can be obtained by biological means such as conventional genetic engineering, and the problem that the genetic sequence of the antibody is easy to lose due to long-term cryopreservation of traditional hybridoma cells is avoided; optimization of the antibody on the gene or protein level is facilitated, and the specificity and affinity of the antibody are further improved.

The invention discloses a sebastes schlegeli testis cell line as well as a construction method and application thereof, and belongs to the technical field of marine fish cell culture. Primary culture and subculture are carried out on sebastes schlegeli juvenile fish testis tissues, so that the sebastes schlegeli testis cell line capable of being continuously subcultured in vitro for a long time is established. The sebastes schlegeli testis cell line established by the invention is in a fibroblast shape, has a good growth curve, and has relatively strong multiplication capacity. The lentiviral vector can significantly express the stromal cell marker gene temp2, can transfect the cell line, and can be subsequently applied to research of gonad development related functional genes. At present, the cell line has been subjected to in-vitro continuous passage for more than 60 generations, can be subjected to cryopreservation, and is high in recovery survival rate. Establishment of the cell line provides a stable and excellent experimental material for research of fish gonad development genes.

The invention relates to the technical field of NK cell culture, and discloses a serum-free medium for NK cells from umbilical cord blood after cryopreservation and a culture method thereof.The medium comprises a coating solution, an induction medium and an amplification medium; wherein the coating liquid is prepared from an anti-CD16 monoclonal antibody and DPBS (diphenylphosphonic acid); the induction culture medium comprises a basic culture medium, a human-derived platelet lysate and cell factors IL-15, IL-18 and IL-21; the amplification culture medium comprises a basic culture medium, a human-derived platelet lysate and IL-2. The culture method comprises the following steps: obtaining mononuclear cells from fresh umbilical cord blood, and then cryopreserving and resuscitating; and carrying out multiplication culture on the recovered umbilical cord blood mononuclear cells by using the serum-free culture medium. According to the NK cell culture medium, the NK cells with high purity, high killing activity and high cytotoxic activity can be obtained without autoserum culture.

The invention relates to a preservation method and application of panax notoginseng germplasm resources, and relates to the technical field of panax notoginseng germplasm preservation. The preservation method comprises the following steps: (1) suspending pseudo-ginseng embryogenic calluses and a WPM culture medium, and screening and collecting to obtain pseudo-ginseng embryogenic cell clusters with the size of 0.45-1.5 mm; (2) transferring the panax notoginseng embryogenic cell cluster into a cryopreservation tube, and adding a loading solution for loading treatment to obtain a loaded embryogenic cell cluster; (3) carrying out vitrification treatment on the loaded panax notoginseng embryogenic cell cluster and a vitrification solution in a cryopreservation tube to obtain a vitrified panax notoginseng embryogenic cell cluster; (4) placing the cryopreservation tube filled with the vitrified pseudo-ginseng embryogenic cell cluster in liquid nitrogen for preservation; and after preservation, carrying out recovery culture on the pseudo-ginseng embryogenic cell cluster. According to the method, the survival rate reaches 90% after preservation, pseudo-ginseng plants can be rapidly differentiated, the differentiated pseudo-ginseng plants survive after being transplanted under natural conditions, the transplanting survival rate reaches 90%, and the pseudo-ginseng plants grow well in the later period.

The invention discloses a micro-fluidic chip for vitrification cryopreservation of oocytes or embryos and a preparation method and application of the micro-fluidic chip. The micro-fluidic chip comprises a micro-fluid channel layer, and the micro-fluid channel layer is arranged in the micro-fluid flowing direction; the microfluid channel layer comprises one or more solution inflow channels, a snakelike mixing channel, one or more shunt channels, a snakelike channel, a cell storage chamber and a waste liquid discharge channel which are communicated in sequence; wherein a first micro-column array is arranged at an outlet of the cell storage chamber, and a second micro-column array is arranged in a channel outside an inlet of the cell storage chamber; and a cell channel is communicated between the cell storage chamber and the second micro-column array. The micro-fluidic chip can realize the integration of operations such as vitrification cryopreservation and rewarming, reduces the cell transfer frequency, avoids pollution, improves the treatment efficiency, can significantly improve the survival rate of oocytes or embryos after rewarming, and can further promote the development of an assisted reproduction technology.

The invention discloses a processing method capable of shortening the fermentation period and reducing the biogenic amine content for inoculated fermentation of smelly mandarin fish, and belongs to the field of food processing. The method comprises the following steps: (1) pretreating mandarin fish; (2) preparing fermentation liquor, wherein the fermentation liquor is prepared from NaCl, Chinese prickly ash, a leavening agent and water; the mass percentage concentration of each component is as follows: 0.1% of pepper, 4-8% of NaCl, 0.5-1.5% of a leavening agent and the balance of water; (3) fermentation process of smelly mandarin fish: completely immersing the fish body in the fermentation liquid, respectively fermenting for 6-12 days at the temperature of 5-20 DEG C, and collecting a smelly mandarin fish sample; and (4) performing vacuum packaging: performing vacuum packaging on the collected smelly mandarin fish samples, and performing cryopreservation at-18 DEG C. The leavening agents are lactobacillus sake, pediococcus pentosaceus, lactobacillus plantarum and mixed strains (lactobacillus sake, pediococcus pentosaceus and lactobacillus plantarum in a ratio of 1: 1: 1) respectively. The fermentation period of the siniperca chuatsi is shortened through inoculated fermentation, the content of biogenic amine is reduced, and the method has very high application and popularization value.

The utility model relates to the technical field of biological sample cryopreservation equipment, in particular to a conveying device for biological sample cryopreservation, which comprises a support plate and a support truss arranged on the support plate, a horizontally-arranged conveying module is arranged on the supporting plate, and a pushing module used for pushing the test tube boxes to the cryopreservation frame is arranged on the conveying module; the material taking mechanism comprises a clamping jaw air cylinder, and clamping blocks are arranged at the two movable ends of the clamping jaw air cylinder. The pushing module comprises a material carrying frame fixedly arranged on the conveying module, the material carrying frame is provided with a test tube box containing position, and a pair of receding grooves used for containing the clamping blocks are formed in the test tube box containing position. Under the action of the receding groove, the test tube box is placed on the material carrying frame in a collision-free mode, the situation that after the clamping blocks loosen the test tube box, the test tube box falls and collides with the material carrying frame, and consequently the biological sample leaks is avoided, and the conveying stability of the biological sample is improved.

The invention provides a Wujin pig seminal plasma protein crude extract and application of the Wujin pig seminal plasma protein crude extract in an anti-freezing agent and an anti-freezing diluent for freeze preservation of Wujin pig seminal fluid. The Wujin pig seminal plasma protein crude extract is derived from Wujin pig seminal plasma, does not contain other exogenous bioactive substances, has a better freezing protection effect on sperms of Wujin pigs and is safer, and the acrosome integrity rate of frozen sperms of the Wujin pigs can be remarkably increased. According to the present invention, with the anti-freezing diluent, the vitality and the movement performance of the cryopreserved Wujin pig sperm can be significantly improved, the active oxygen level is reduced, the mitochondrial membrane potential and the energy metabolism are improved, and the oxidation resistance is enhanced, such that the vitality of the Wujin pig sperm can be easily improved, and the mitochondrial function can be easily maintained; the research provides key technical support for establishment of a Wujin pig semen cryopreservation technology, and has great practical significance for Wujin pig breeding and population propagation.

The invention provides a technology for preparing universal DNT cells. In the invention, the prepared raw materials are derived from'white filtering residues' after white filtering treatment is carried out to avoid adverse blood transfusion reaction, and the DNT cells extracted from the white filtering residues can be a mixed culture of one or more healthy donors. The universal DNT cell obtained by the method has the characteristics of high yield, high purity, high in-vitro tumor killing activity, suitability for large-scale production and the like. The initial raw material can be fresh cells or re-thawed cells after cryopreservation, the raw material source bottleneck of large-scale preparation of cell drugs is fundamentally solved, the value of leukocytes filtered out in the production process of blood products is re-excavated, and great medical value and commercial application prospects are achieved.

The invention discloses a biological sample low-temperature preservation box, and particularly relates to the technical field of DNA sample preservation, the biological sample low-temperature preservation box comprises a box body and a first box door, the first box door is movably connected with the box body through a hinge, a touch screen, a first camera and a fingerprint lock are fixedly installed at the top end of the box body, and a second camera is fixedly installed on the front side of the box body; a semiconductor refrigerator is fixedly installed in the box body, a second box door is movably arranged on the rear side of the first box door, and the second box door is movably connected with the front side of the box body through a hinge. The threaded ice storage barrel is rotated to be far away from the box body, then ice blocks are placed in the threaded ice storage barrel, finally the threaded ice storage barrel is installed and fixed, auxiliary refrigeration is conducted on the interior of the box body, and meanwhile, along with gradual melting of the ice blocks, generated water enters the water storage barrel through the guide pipe; the one-way valve can prevent water in the water storage barrel from flowing back into the threaded ice storage barrel, the structure is simple, and auxiliary refrigeration can be conveniently carried out on the interior of the box body.

The utility model relates to the technical field of picking and moving of biological sample cryopreservation tubes, and discloses a cryopreservation tube anti-falling structure which comprises a tube picking assembly, an anti-falling movement assembly, a fixing assembly and a control module, and the tube picking assembly and the anti-falling movement assembly are both installed on the fixing assembly. The pipe picking assembly is composed of a cryopreservation pipe, a suction head, a suction head spring, a linear bearing and an air pipe connector. The cryopreservation tube picking assembly is arranged, the cryopreservation tube is sucked through negative pressure, the cryopreservation tube is blown away through positive pressure, the cryopreservation tube picking device is simple in structure, few in component parts, small in size, higher in adaptability, low in manufacturing cost and stable and reliable in operation, the anti-falling movement assembly is arranged, two clamping jaws are pressed through a hinge pin spring to clamp the cryopreservation tube, and the cryopreservation tube is prevented from falling accidentally; the cryopreservation tube clamping device is simple in structure, low in manufacturing cost and stable and reliable in operation, the cryopreservation tube can be clamped to be prevented from falling off even if power failure or air leakage occurs due to the fact that the spring is used for pressing the two clamping jaws to clamp the cryopreservation tube, and maintenance is convenient.

The invention discloses a method for cryopreserving mature dendritic cells, and the method for cryopreserving the mature dendritic cells comprises a dendritic cell cryopreservation solution. The dendritic cell cryopreservation liquid is mainly prepared from the following raw materials in percentage by volume: 15 to 16v / v percent of human albumin injection, 1 to 3v / v percent of antifreeze protein liquid, 17 to 20v / v percent of compound dextran 40 injection, 17 to 20v / v percent of compound electrolyte injection and 41 to 50v / v percent of CryoStorCS10, in addition, the cryopreservation liquid also comprises Setanaxib, and the concentration of the Setanaxib in the cryopreservation liquid is 20 [mu] mol / L. The cryopreservation liquid is used for cryopreservation treatment of the mature dendritic cells, through synergistic interaction among different components in the cryopreservation liquid, the recovery survival rate of the cryopreserved mature dendritic cells is larger than or equal to 80%, and the antigen presentation capacity of the mature dendritic cells can be preserved to a large extent.

The invention provides a preparation method of Al18F tracer integrated freeze-dried powder, a kit and application thereof, and the preparation method comprises the following steps: dissolving 1-10 mg of a precursor by using deionized water, adding a pH buffer solution with the concentration of 0.2-0.5 M and an AlCl3 solution with the concentration of 10 mM, with the ratio of the amount of substance of AlCl3 to the amount of substance of the precursor being 0.8-1.0, then continuously adding a stabilizer solution with the concentration of 0.5 M and 1-4 mg of an excipient, and uniformly stirring to obtain the Al18F tracer integrated freeze-dried powder. And finally, adding deionized water to enable the concentration of the precursor to be 0.5-1.0 mM, uniformly mixing the solution, sub-packaging the solution into 1.5 mL cryopreservation tubes or 2 mL penicillin bottles according to a certain volume, and then putting the cryopreservation tubes or the penicillin bottles into a vacuum freeze dryer to remove water at a low temperature to obtain powder or freeze-dried cakes, and the container is a freeze-dried powder / cake storage container and is used for storing the freeze-dried powder / cakes. Meanwhile, the reaction container is also used during radioactive labeling.

The utility model discloses a biological sample low-temperature preservation device. A freezing storage set and a rotating assembly are placed in a low-temperature storage cavity; the bottom of the lifting part is arranged at the top, located in the transition area, of the heat preservation tank body, a lifting assembly is arranged at the top of the lifting part, and a lifting part of the lifting assembly can sequentially penetrate through the heat preservation taking and placing part and the transition part and extend into the heat preservation part; the first liquid level sensor, the liquid inlet pipeline, the first temperature sensor and the second liquid level sensor are all located in the heat preservation tank body, the first liquid level sensor and the liquid inlet pipeline are both close to the lower surface of the rotating disc, the first temperature sensor is located on the peripheral wall face of the rotating disc, and the second liquid level sensor is located on the inner top of the heat preservation tank body. The liquid inlet valve is communicated with the liquid supplementing valve, the liquid supplementing valve is communicated with the liquid inlet pipeline, the exhaust valve is communicated between the liquid supplementing valve and the liquid inlet valve, and the exhaust valve is communicated with the vertical channel; the second temperature sensor is located in the cylinder. When the cryopreservation box is taken and placed, the cryopreservation box is in a low-temperature area of-80 DEG C to-170 DEG C, so that the biological sample is always in a protection state.

The disclosure provides for cryopreservation formulations, cryopreservation media and method of cryopreserving mammalian cells or tissues. The cryopreservation formulations and media contain a combination of cryoprotective agents selected from the group consisting of; (a) glycerol and trehalose; (b) glycerol, trehalose, and ascorbic acid; (c) glycerol, trehalose, and taurine: and (d) glycerol, trehalose, ascorbic acid, and taurine. In certain embodiments, the cryopreservation formulations and media are free of dimethyl sulfoxide.

The invention discloses a ready-to-use activated human umbilical cord mesenchymal stem cell as well as a preparation method and application thereof. The preparation method comprises the following steps: carrying out passage on P0-generation cells for three times to obtain P3-generation cells; cryopreserving P3 generation cells, and establishing a cell seed bank; resuscitating the cells in the seed bank for three times of passage to obtain P6-generation cells; and carrying out activation culture on the P6-generation cells, and resuspending the activated P6-generation cells in a cell protective agent to obtain the human umbilical cord mesenchymal stem cell original suspension. The invention also discloses an umbilical cord mesenchymal stem cell cryopreservation preparation. The cryopreservation preparation is obtained by mixing the human umbilical cord mesenchymal stem cell original suspension with a cryopreservation protective agent, sub-packaging and cryopreserving. Experiments prove that the human umbilical cord mesenchymal stem cell cryopreserved preparation prepared by the invention can be stored for a long time, is convenient to store, transport and use clinically, can complete effectiveness and safety index detection within the shelf life, and has the same activity as a fresh preparation.

The present disclosure relates to compositions and methods for cryopreservation of mitochondria, cryopreserved compositions comprising mitochondria, and therapeutic uses thereof.

The present disclosure provides methods and devices for inhibiting or reducing biological contaminants in biological matters during long term preservation while maintaining desired properties of the biological matter by applying the biological matter specific long term isochoric freezing or mild isochoric freezing treatment parameters.

The invention belongs to the technical field of bioengineering, and particularly discloses an in-vitro amplification method of canine NK cells capable of efficiently killing tumor cells and application of the canine NK cells. 3 mL of dog peripheral blood is taken, and dog peripheral blood mononuclear cells PBMC are obtained through separation; activating, amplifying and culturing canine NK cells capable of efficiently killing tumor cells; and collecting the amplified canine NK cells capable of efficiently killing the tumor cells, and storing the canine NK cells in a cryopreservation solution according to the cell density of 1 * 10 < 8 > cells / mL. According to the method, the canine NK cell with high amplification multiple can be obtained by only needing 3mL of canine peripheral blood, and the amplification multiple is high; the number of the dog NK cells of the healthy volunteer dog sample is 3.02 * 108, the amplification multiple is 1008, the total number of the dog NK cells of the tumor volunteer affected dog sample reaches 6.84 * 107, and the amplification multiple is 340; and the canine NK cell obtained by the invention has the effect of efficiently killing tumor cells.

The invention provides a preparation method of a skull for autologous replantation, and relates to the technical field of skull treatment. The method comprises the following steps that the taken-out autologous skull is subjected to ultraviolet radiation and then conveyed into a clean room to be subjected to photographing treatment; sequentially carrying out iodophor soaking, alcohol soaking and normal saline flushing on the skull, and then carrying out surface tissue impurity removal and cleaning on the skull; putting the treated skull into a packaging bag, adding a cryopreservation solution to wrap the skull, and then performing vacuumizing and sealing treatment; and sterilizing and cooling the skull subjected to the sealing treatment, and finally feeding the skull into a bone library for cryopreservation. According to the method, the skull of a patient is taken out after an operation to serve as an autologous bone raw material, so that the skull has bone activity within a certain period of time, finally, the skull is replanted to the skull of the patient, the body burden and conflict psychology of the patient are reduced, the exclusive transportation, treatment and storage modes are adopted, the rejection of the patient is reduced, the risk of bone resorption is reduced, and the bone activity is guaranteed; the replanting rate is increased.

The utility model relates to the technical field of sample preservation, and discloses a quick storing and taking device for a liquid nitrogen tank cryopreservation box, which comprises a machine body, support frames are fixedly connected to the periphery of the outer surface of the machine body, a mounting frame is fixedly connected to one side in the machine body, and a movable door motor is fixedly mounted on one side of the mounting frame through bolts. According to the utility model, the whole equipment is arranged, the access of the cryopreservation box is controlled through the moving door motor, the operation can be quickly and accurately completed, the device is particularly suitable for processing a large number of samples, the working efficiency is obviously improved, the labor intensity is reduced through the electric operation mode, the physical output of personnel when the samples are frequently accessed is reduced, and the labor intensity is reduced. Through cooperation of the jacking motor and the moving door lead screw, the cryopreservation box can be adjusted in a self-adaptive mode, it is ensured that the cryopreservation box is safely and stably stored, the cryopreservation boxes of different layers can be conveniently stored and taken out, the cryopreservation box can be rapidly moved into or out of the liquid nitrogen tank through combination of the conveying motor, the transmission shaft and the conveying belt, and the working efficiency is improved.

The present disclosure provides methods for re-stimulating TIL populations that lead to improved phenotype and increased metabolic health of the TILs and provides methods of assaying for TIL populations to determine suitability for more efficacious infusion after re-stimulation.

The invention belongs to the technical field of biological culture, and particularly relates to a chlorella extracellular fluid and a preparation method and application thereof.The preparation method of the chlorella extracellular fluid comprises the following steps that a BG11 culture medium is inoculated with a chlorella solution cultured to the logarithmic phase, oxcarbazepine is added, and the chlorella extracellular fluid is obtained; the preparation method comprises the following steps: placing chlorella in an intermittent illumination environment with the temperature of 20-30 DEG C and the rotating speed of 100-200 rpm for growth, centrifuging the chlorella to obtain supernate after the chlorella grows to the 10th-16th day, filtering, sub-packaging, and performing cryopreservation to obtain the chlorella extracellular fluid. By adding oxcarbazepine, the chlorella extracellular fluid prepared in a specific culture medium and a specific growth environment has high oxidation activity, excellent stability and excellent degradation capacity, and a new thought is provided for solving the pollution treatment problem of current environmental pollutants such as pharmaceutical products, polycyclic aromatic hydrocarbons and dyes.

The present invention provides an extracellular vesicle protectant and its application. The extracellular vesicle protectant has a simple composition, including a buffer system, a saccharide substance, and a surfactant. It can be used as a cryopreservation solution or a lyophilization protectant, and has a wide applicable temperature range. The protectant can reduce the storage and transportation cost of exosomes and expand the applicable range of exosomes. The exosome lyophilized powder prepared by using the above protectant and exosomes can be stably stored for a long time. The lyophilized powder can be realized on a large-scale lyophilization production device, making it possible to industrially produce exosomes on a large scale.

The utility model relates to the field of experimental consumables, in particular to a cryopreservation tube protective cover and a cryopreservation tube. The cryopreservation tube protection cover is matched with a tube body and a cover body of a cryopreservation tube and comprises a shell and a separation membrane. The housing has a first section and a second section opposite the first section. The first section of the shell is matched with the pipe body and connected with the pipe body in a sealed mode. And the second section of the shell is matched with the cover body and is detachably connected with the cover body. The edge of the separation film is connected to the inner wall of the shell and used for closing the opening of the pipe body. The cryopreservation tube comprises the tube body, a cover body and a protective cover. Whether the cryopreservation tube is opened for the first time or not is judged by judging whether the separation membrane is punctured and damaged or not, and the uniqueness function of a biological sample can be achieved easily. A sample is protected by the separation membrane after the cover body is opened and before sampling, so that the cleanliness of the sample is improved.

The utility model discloses a simple freezing heat preservation device for a cryopreservation box, which belongs to the technical field of cryopreservation and comprises a base, a box frame and a cover. A concave cavity is formed in the top surface of the base; the box frame is located in the concave cavity and comprises a box frame body, the top face of the box frame body is provided with a cryopreservation box containing position used for containing a cryopreservation box, the bottom face of the box frame body is fixedly connected with the base through a plurality of supporting columns, and the box frame body is further provided with hollow holes opposite to the cryopreservation box containing position; and the cover is detachably and fixedly connected to the top surface of the base so as to seal the concave cavity. According to the utility model, the cover state of dry ice on the cryopreservation box can be effectively kept in the sample transfer process, and the sample freezing effect is ensured not to be influenced by jolt.

The invention discloses a method for establishing a cell line by serum-free culture of PGCs (Primordial Germ Cells) of breeding poultry, the cell line is prepared by blending calcium-free DMEM (Dulbecco Modified Eagle Medium), water and Knockout-DMEM, the cell line is more suitable for osmotic pressure and calcium ion concentration required by proliferation of the PGCs of the breeding poultry, various components of serum-free culture are perfected by using fresh egg white, and the cell line is more suitable for proliferation of the PGCs of the breeding poultry. The combination of growth factors required by primordial germ cells is optimized by adding the FGF-1, so that 100% of cell line establishment efficiency and rapid growth of cells from reproduction crescent, blood and gonad tissues of chicken embryos are realized, and a technical basis is provided for poultry frozen breed conservation, stem cell propagation and gene editing operation.

The invention provides an NK cell cryopreservation resuscitation method and an NK cell cryopreservation resuscitation kit, and relates to the technical field of biology. The cryopreservation recovery method of the NK cells, provided by the invention, comprises the following steps: pre-treating the NK cells to be cryopreserved in an NK cell in-vitro amplification culture medium containing 0.1-50mM of NAM and 1-1000ng / mL of recombinant human IL-18; the pretreated NK cells are subjected to cryopreservation in an NK cell cryopreservation solution containing 1-50 mM of NAM; and resuscitating the cryopreserved NK cells to obtain resuscitated NK cells. According to the cryopreservation recovery method, the survival rate, the recovery rate and the functional activity of NK cell recovery for 24 hours can be remarkably improved, and possibility is provided for NK cell seed bank construction, gene editing and spot type product development.

The invention discloses a method for extracting mesenchymal stem cells from urine. The method comprises the following steps: (1) sampling and inoculating urine; (2) amplification and passage of mesenchymal stem cells; (3) identifying the urine stem cells; and (4) a cryopreservation and resuscitation method of the urine stem cells. And performing biological phenotype identification according to the standard formulated by the International Cell Therapy Association to confirm the mesenchymal stem cells. The method is low in cost, simple and rapid, and the used culture solutions are all commercialized and are very convenient to purchase and use. The culture solution special for the stem cells is used in the first two weeks of the extraction period, after passage, when enough stem cells are obtained, the culture medium can be replaced with a common DMEM culture medium, and the manufacturing cost is greatly reduced. And secondly, the materials are simple, rapid and non-invasive, and the sample of the method for extracting the mesenchymal stem cells from urine comes from a patient, so that the problem of heterogeneity of cell sources is reduced. And a foundation is laid for stem cell treatment and gene editing treatment of genetic diseases and major difficult diseases.

The invention discloses a construction method of a human epidermal organoid, and belongs to the technical field of organoid cell culture. The method is mainly used for optimizing a culture method of human epidermal organoid, and particularly relates to an efficient epidermal organoid culture technology based on a specific culture medium and an enzyme digestion system. According to the method, a # imgabs0 # human epidermal organ complete culture medium is adopted for preparation, and a basic culture medium and an additive are mixed to form the complete culture medium suitable for cell growth. The primary specimens are treated, and the steps of cleaning, digestion, cell collection and the like are included, so that the activity and functions of cells are ensured. Through accurate inoculation and culture conditions, the formation and growth of the organoid are promoted, and the culture in a proper environment is ensured, so that the formation efficiency of the organoid is improved. The organoid is subjected to passage and cryopreservation treatment, so that the activity and functional stability of the organoid in the long-term storage and use process are ensured.