246 results about "Cell extraction" patented technology

The invention belongs to the technical field of inspection and extraction, and particularly relates to an automatic T lymphocyte suction device which comprises an automatic suction device body, the automatic suction device body comprises a bearing table, a rotating disc with a circumferential rotating function is arranged on the bearing table, and a plurality of centrifugal assemblies are evenly distributed on the rotating disc. A centrifugal tube feeding assembly facilitating feeding of centrifugal tubes is arranged in front of one side of the bearing table, a tube carrying and conveying assembly is arranged on the portion, located on one side of the centrifugal tube feeding assembly, of the bearing table, a tube feeding mechanism is arranged at the end of the conveying belt mechanism, and a reaction reagent adding assembly is arranged on one side of the tube carrying and conveying assembly. A sample feeding assembly is arranged on one side of the upper portion of the workbench, and a first suction assembly is arranged above the multi-direction adjusting assembly. According to the invention, multiple samples can be processed in batches, the working efficiency and the cell extraction purity are improved, the influence of manual operation on the result is reduced, the accuracy of the detection effect is ensured, and the use requirements are met.

The present disclosure relates to the seminal discovery of a generation and use of genetically engineered Vibrio sp. Provided is the use of the genetically engineered bacteria for the construction, maintenance, manipulation, and / or propagation of DNA constructs; protein expression; protein secretion; vectors and other metabolic tools; metabolic engineering; expression of cellular extracts for cell-free biology; shuttle vectors; cloning vectors; and for synthetic biology applications. The disclosure also relates to the use of the replication machinery of Vibrio sp. as a cloning or expression vector for replication of recombinant DNA constructs. The disclosure also relates to methods of use of the above.

The invention relates to a method for rapidly detecting the cell adhesiveness of probiotics and application of the method. The method comprises the following steps: adding the probiotics into adherent culture cells, incubating, discarding liquid, digesting and collecting the cells by pancreatin, extracting total DNA (Deoxyribonucleic Acid), performing qPCR (Quantitative Polymerase Chain Reaction) amplification, and calculating the adhesion quantity of the probiotics on a single cell according to a Ct value. The invention provides a method for rapidly detecting the cell adhesion rate of probiotics by using a qPCR (quantitative polymerase chain reaction) technology, which comprises the following steps of: extracting total genome DNA (deoxyribonucleic acid) after the probiotics are adhered to cells on the basis of the gene expression difference of prokaryotic and eukaryotic cells, and detecting the specific gene expression level of the probiotics and the cells by using the qPCR technology. The cell adhesion ability of the probiotics is evaluated by calculating the ratio of the gene expression quantities of the two, and the method has the characteristics of high efficiency, rapidness, high accuracy and the like.

The invention relates to a cell extraction method and device, computer equipment and a storage medium. The method comprises the following steps: acquiring various cell images of cells to be sorted, and identifying fluorescent dots in each cell image through a deep learning model; determining the fluorescence signal intensity of the fluorescent dots in each cell image; and according to the fluorescence signal intensity of the fluorescent dots in the multiple cell images, absorbing cells meeting a preset sorting condition from the to-be-sorted cells. By adopting the method, the cell extraction accuracy can be improved.

The invention provides a stem cell extract composition for promoting hair growth and a preparation method thereof, and belongs to the technical field of cosmetics. Fetal calf serum, dexamethasone, penicillin, streptomycin, interleukin-2 and magnetic composite particles are added into a DMEM culture medium to prepare the culture medium, the culture medium is used for in-vitro culture of stem cells, during culture, a stem cell solution is prepared by combining a low-frequency magnetic field with low-energy laser culture, exosomes are extracted from an extracting solution, the exosomes and the stem cells are mixed and embedded to prepare embedded microspheres, and the embedded microspheres are used for in-vitro culture of the stem cells. The stem cell extract composition for promoting hair growth is prepared by uniformly mixing the stem cell extract with extracts extracted from fresh ginger, ginseng and polygonum multiflorum, the storage stability of stem cells and exosomes is obviously improved, hair growth is promoted, hair follicle development is promoted, mouse hair is induced and maintained in a growth period, and the stem cell extract composition is free of hormone addition, safe and non-irritant, and has a good application prospect. Wide application prospects are realized.

The utility model discloses a thyroid needle biopsy device, including thyroid needle biopsy device main part, first locking piece, second locking piece and cushion pad, the inner wall of alcohol tube is movably connected with push component, the bottom of spray tube inner wall is fixedly connected with dispersion component. Through the arrangement of the mounting plate, the alcohol pipe and the spray pipe, the alcohol pipe can be fixed and limited by the mounting plate, the alcohol pipe is used for storing disinfection alcohol, and the disinfection alcohol can be sprayed to the skin of a patient through the spray pipe, so that the problem that the skin of the patient needs to be disinfected by alcohol before thyroid tissue cells are obtained is solved; however, the problems that an existing thyroid needle biopsy device body does not have a disinfection function, an alcohol disinfection device needs to be additionally prepared in advance, great inconvenience is caused, and the efficiency and the quality of cell extraction are affected due to the fact that the outer skin of a cell extraction area is not accurately disinfected manually are solved.

The invention belongs to the technical field of cell extraction, and particularly relates to a method for extracting cells by enzymolysis of vitreous bodies, which comprises the following steps: centrifuging a vitreous body sample at room temperature, discarding supernate, and adding digestive enzyme into precipitate; after adding digestive enzyme, digesting in a water bath for 30 minutes; and after the enzymolysis is finished, centrifuging the enzymatic hydrolysate at room temperature, discarding the supernate, and collecting the precipitate, namely the enzymolysis vitreous cells. According to the technical scheme, a simple and efficient vitreous body enzymolysis method is provided, and a large number of cells can be provided for follow-up research.

The present invention relates to a cartilage tissue single cell extraction kit and an extraction method thereof, the kit comprises: (1) a first enzyme for enzymatic hydrolysis, the first enzyme for enzymatic hydrolysis comprising one or more of trypsin, chondroitinase ABC and fucosidase; (2) a second enzyme for enzymolysis, wherein the second enzyme for enzymolysis comprises collagenase II, neutral protease II, hyaluronidase and Dnase I; and (3) a basic solution, wherein the basic solution comprises a basic culture medium and / or a buffer solution. The invention also provides a method for extracting single cells from cartilage tissues by using the kit. The cartilage tissue single-cell suspension prepared by the method has the advantages of high cell yield, high activity and the like, and has a wide application prospect.

The invention provides a lysosome extraction method. The lysosome extraction method comprises the following steps: (1) extracting cells; (2) tissue extraction; (3) crude separation of lysosome; and (4) separating the lysosome by magnetic beads. Through EDTA-free digestion optimization, lysis solution and buffer solution formula optimization, a filter membrane interception-magnetic bead purification combined technology, a double-antibody synergistic capture strategy and a competitive mild elution system, the four core problems of low purity, large activity damage, tedious operation and strong equipment dependence in a traditional lysosome separation technology are systematically solved. Experimental data show that the lysosome yield and functional integrity of the method are obviously superior to those of a density gradient centrifugation method, and the method is suitable for trace samples and conventional equipment and has high conversion value. In the future, the technical barrier can be further optimized through automation equipment integration and multi-dimensional function verification, and the application of the method in basic research and clinical diagnosis is promoted.

The invention belongs to the technical field of antibody preparation, and particularly relates to an anti-NKG2D nano antibody as well as a preparation method and application thereof. The method comprises the following steps: constructing and expressing NKG2D protein, immunizing alpaca to obtain immune alpaca PBMC (peripheral blood mononuclear cells), extracting RNA (ribonucleic acid) of the PBMC, carrying out reverse transcription to obtain cDNA (complementary deoxyribonucleic acid), obtaining an antibody gene segment through PCR (polymerase chain reaction), and constructing the antibody gene segment on a phage expression vector; the method comprises the following steps: carrying out electrotransformation on a vector with an antibody gene segment to an SS320 electrotransformation competent state to obtain an NKG2D immune antibody library, carrying out two rounds of NKG2D protein panning and Clone ELISA detection to obtain an antibody combined with NKG2D protein, carrying out sequencing analysis, selecting a unique sequence, and carrying out high-throughput expression through a mammalian cell high-throughput expression system to obtain an antibody protein. And carrying out FACS detection by using an NKG2D high-expression cell line to finally obtain the antibody combined with the NKG2D. The method disclosed by the invention is simple and quick to operate; a mammalian cell expression system induces high-efficiency expression of the antibody, processing modification can be performed after translation, and the activity of the antibody is closer to that of a natural antibody.

The system for dismantling the spent lithium-ion battery pack includes: a transfer section including transfer rails and a gantry robot on the transfer rails; a discharge processing section including an index table on which a discharging task of discharging a battery pack is performed; a battery disassembly section including a worktable on which a disassembly task is performed to cut connections of the battery pack, disassembly robots, and first transfer trolleys; and a cell extraction section including an extraction robot, a second transfer trolley configured to carry out battery cells, of which the connections have been cut by the disassembly task, to the outside, and a third transfer trolley configured to carry out a tray on which the battery cells are placed to the outside. The index table, the worktable, the second transfer trolley, and the third transfer trolley are arranged directly below the transfer rail.

The invention discloses a genome sequencing sample processing method based on a microdissection technology. The method comprises the following steps: (1) preparing a paraffin section or a frozen section; (2) carrying out HE staining on the section obtained in the step (1); (3) carrying out laser microdissection on nucleic acid; (4) fragmenting the DNA sample, and treating a product obtained in the step (3) by utilizing an ultrasonication method to obtain a DNA fragment; a non-contact ultrasonic crusher is adopted, the power is 80%, ultrasonic treatment is carried out for 10 s, stopping is carried out for 20 s, and 15 cycles are carried out. And (5) carrying out on-machine pretreatment on the genome sequencing sample, wherein the treatment comprises tail end repair and joint connection treatment. According to the method disclosed by the invention, accurate cutting of the sample can be realized, a DNA sample is extracted from a very small amount of cells and a sample for sequencing is constructed, the sample can achieve the same or higher sequencing data quality as that of a traditional method, the usage amount of the cells is greatly reduced, and the repeatability of an experiment is improved.

The invention discloses a preparation method and application of a porous GelMA microneedle for delivering AIE molecules based on HSF derived exosome. The preparation method comprises the following steps: S1, preparing TTMN molecules; s2, dispersing DPPC and Chol in chloroform, performing water bath evaporation at 50 DEG C to form a thin film, hydrating the thin film in PBS, incubating to enable DPPC and Chol to form a liposome structure again, and uniformly dispersing TTMN molecules in the liposome structure to obtain TL; s3, extracting an exosome from the HSF cells; s4, loading the TL into the exosome by utilizing an electroporation technology, so as to obtain TE; s5, preparing the GelMA; and S6, mixing the GelMA with the TE, so that the TE is uniformly dispersed in the GelMA to obtain a mixed solution, injecting the mixed solution into a microneedle mold, and irradiating with 405nm laser to obtain the porous GelMA microneedle. According to the invention, high-efficiency treatment of hypertrophic scars is realized, and the system has a remarkable drug delivery effect, an innovative photodynamic therapy, a high-efficiency scar tissue remodeling effect and good biological safety and stability, and has important clinical application value and market prospect.

The invention relates to the technical field of stem cell extraction, in particular to a stem cell extraction process, solves the problems that when stem cells in a placenta are extracted in the prior art, the temperature in the placenta treatment process is inconvenient to keep stable, and the placenta is inconvenient to fully crush and clean, and is used for extracting the placenta stem cells. The stem cell extraction process is realized through stem cell extraction equipment, the stem cell extraction equipment comprises a circulating cooling mechanism, a coarse crushing mechanism, a fine crushing mechanism and a digestion separation mechanism, the coarse crushing mechanism is mounted on the inner side of the upper end of the circulating cooling mechanism, the fine crushing mechanism is mounted at the bottom end of the coarse crushing mechanism, and the digestion separation mechanism is mounted at the bottom end of the fine crushing mechanism. A digestion and separation mechanism is mounted on one side of the fine crushing mechanism, and the circulating cooling mechanism comprises a supporting base. Through continuous cleaning and gas injection in the stem cell extraction process, the temperature stability of the stem cells can be effectively kept, and the placenta cleaning and crushing effects can be conveniently improved.

The invention relates to an extraction and culture method of urine-derived stem cells, and belongs to the field of cell biology and regenerative medicine. The method comprises the following steps: coating a culture plate with a gelatin solution; collecting clean midstream urine of healthy people and adding antibiotics; centrifugally separating the cell precipitate; washing with a phosphate buffer solution; re-suspending the precipitate by using a urine stem cell complete culture medium for primary culture; and after the cells grow to a certain fusion degree, carrying out passage amplification. The urine stem cell complete medium comprises a basic medium, fetal calf serum, epidermal growth factors, platelet-derived growth factors and basic fibroblast growth factors. By adopting the method disclosed by the invention, the urine-derived stem cells can be efficiently and stably separated from urine, and the obtained cells have high multiplication capacity, good stemness maintaining capacity, multidirectional differentiation potential and immunoregulation function, and can provide an ideal seed cell source for treating diseases such as diabetic erectile dysfunction.

The invention discloses a multi-enzyme preparation for industrial cell dissociation as well as a use method and application thereof, and belongs to the technical field of extraction of seed cells for industrial culture of meat. According to the invention, a multi-enzyme preparation composed of a component 1 and a component 2 is adopted; the component 1 comprises neutral protease, hyaluronidase and elastase; and the component 2 comprises matrix metalloproteinase, papain and plasmin. The invention also discloses a preparation method and application of the multienzyme preparation. The reasonable enzyme ratio and the optimized treatment conditions are beneficial to protecting the integrity of cell membranes, maintaining high cell activity and remarkably improving the dissociation efficiency and the cell recovery amount; the complex enzyme preparation is high in adaptability, can be used for various tissue types and dissociation requirements, and is convenient for standardized and large-scale production; in combination with automatic dissociation equipment, the compound enzyme preparation can realize efficient and stable industrial operation, and provides important technical support and wide application prospects for the fields of meat culture industry, tissue engineering, cell therapy and the like.

The invention discloses an Escherichia coli culture medium and application thereof in preparation of cell extracts required by cell-free protein synthesis, the culture medium comprises an initial culture medium or an initial culture medium and a supplementary culture medium, the initial culture medium comprises fructose, lactose, magnesium sulfate, ammonium sulfate, dipotassium phosphate, monopotassium phosphate, citric acid, trace elements, a vitamin solution and a nitrogen source; the fed-batch culture medium comprises a carbon source and a nitrogen source, the carbon source is fructose and a magnesium sulfate solution, and the nitrogen source comprises a nitrogen source solution added with yeast powder and yeast peptone. According to the culture medium disclosed by the invention, the escherichia coli can maintain logarithmic phase growth and reach relatively high thallus density (OD600 is greater than or equal to 60), and the prepared escherichia coli cell extract is used for cell-free protein synthesis, has high activity consistent with that of a cell extract obtained by low-density fermentation, and can be used for expressing various foreign proteins. According to the invention, the problem of low thallus density during preparation of the cell-free protein synthetic cell extract is solved, and the expression yield of the cell-free protein is effectively improved.

The invention belongs to the technical field of antibody preparation, and particularly relates to an anti-CD38 nano antibody as well as a preparation method and application thereof. The method comprises the following steps: constructing and expressing CD38 antibody protein, immunizing alpaca, obtaining immune alpaca PBMC (peripheral blood mononuclear cells), extracting RNA (ribonucleic acid) of the cells, carrying out reverse transcription to obtain cDNA (complementary deoxyribonucleic acid), carrying out PCR (polymerase chain reaction) to obtain antibody gene segments, constructing the antibody gene segments on a phage expression vector to construct a CD38 immune antibody library, carrying out two rounds of panning, carrying out Clone ELISA (enzyme-linked immuno sorbent assay) detection on clones to obtain an antibody combined with the CD38 protein, carrying out sequencing analysis, and selecting a unique sequence. High-throughput expression is carried out through a mammalian cell high-throughput expression system, antibody protein is obtained, FACS detection is carried out through a CD38 high-expression cell line, and finally the antibody combined with CD38 is obtained. The method is simple and rapid to operate, a mammalian cell expression system induces efficient expression of the antibody, processing modification can be performed after translation, and the activity of the antibody is closer to that of a natural antibody.

The invention discloses an SD rat neuropathic pain model dorsal root ganglion cell extraction method, and relates to the technical field of dorsal root ganglion cell extraction. In order to facilitate further research of physiological functions of dorsal root ganglion cells in in-vitro cells; comprising the following steps: selecting an SD (Sprague Dawley) rat as an experimental animal, carrying out sciatic nerve branch selective ligation (SNI), ligating tibial nerves and common peroneal nerves in sciatic nerves, keeping the completeness of the peroneal nerves, and observing the touch sensitivity change of the outer side area of the rat claw after the operation to verify that the model is successfully constructed. According to the method, the extraction efficiency of the dorsal root ganglion cells is effectively improved through the cell treatment steps such as surgical operation under the sterile condition, reasonable use of digestive enzymes and centrifugation and resuspension, so that the experiment time and cost are saved, the smooth proceeding of subsequent experiments is ensured, and the neuropathic pain model is established, so that the development of the neuropathic pain model is promoted. And a basis is provided for subsequent extraction and research of the dorsal root ganglion cells.

The invention discloses a method for extracting primary hepatocyte lipid of an ICR mouse. The method comprises the following steps: digesting and extracting hepatocytes by using a buffer working solution 2 containing type IV collagenase by using a two-step collagenase perfusion method, then laying rat tail collagen in a pore plate, inoculating the extracted hepatocytes on the pore plate, culturing the cells, and finally extracting lipid from the cultured cells. The method is oriented to the ICR mouse, the problem that a primary hepatocyte extraction and culture system is unstable is solved, the cell extraction time is greatly shortened, the survival rate and the cell activity of the cells are improved, and the ICR mouse primary hepatocyte model is more effectively applied to lipidomics research.

The invention discloses a ship declaration cooperative processing system and method based on a multi-user role, and relates to the technical field of ship declaration, and the method comprises the steps: instantiating a digital twin software object for a new ship declaration request; extracting core semantic assertions, metadata and semantic potential energy parameters, packaging the core semantic assertions, the metadata and the semantic potential energy parameters into a semantic cell data unit, traversing all semantic cells in the digital twin, extracting the core assertions of the semantic cells, carrying out semantic matching on the core assertions and nodes in the domain knowledge graph, finding out a corresponding knowledge graph standard concept node, determining an acting force rule, and carrying out semantic analysis on the acting force rule; all semantic cells are placed in a semantic field, and each cell is endowed with a charge attribute obtained through mapping of the knowledge graph according to the core assertion of the cell; and the system runs a physical simulation cycle until the system reaches a steady state or a quasi-steady state, the total potential energy of the whole cell system is calculated, the total potential energy of the system is compared with a preset threshold interval through a normalization function and a calculated semantic stability score, and a decision branch is automatically triggered.

To make it possible to efficiently separate and extract vascular cells from a tissue mass only without damaging the vascular cells, and easily secure the number of vascular cells.SOLUTION: A cell dispersion apparatus capable of selectively extracting vascular cells 7 from a tissue mass 5 comprises: a net-like mounting stage 12 on which the tissue mass 5 is mounted; an ejection port part 42 of the nozzle head 41 which ejects high-pressure UFB jet water running water 43 to the tissue mass 5 mounted on the mounting stage 12, and separates the vascular cells 7 from the tissue mass 5; a high-pressure pump part 22 which generates the high-pressure UFB jet water running water 43 ejected from the ejection port part 42; and an ultrahigh pressure pump part 23 which ejects UFB jet water running water 44 of higher pressure to a blood vessel 6 being a residue of the tissue mass 5, to which the UFB jet water running water 43 was ejected, by the ejection port part 42 of the nozzle head 41.SELECTED DRAWING: Figure 1

The present invention is related to the field of tissue regeneration. It concerns more particularly new processes, tubes and devices for thrombin, platelet concentrate and wound healant preparations, alone or in combination with cell extracts, cell compositions and uses thereof.

The application relates to the technical field of biological sample pretreatment, and discloses a high-efficiency separation method of a tissue cell extraction device. The method comprises the following steps: performing parameter calibration and reference loss modeling of a driving motor, and eliminating background interference of an actuator; applying a disturbance signal to a dissociation chamber, locking the characteristic resonance frequency of a tissue sample through frequency domain energy analysis, and performing pulsating crushing to accelerate the dissociation of a matrix structure; extracting the net torque of the motor in real time, deducing the fluid dynamic viscosity, adaptively adjusting the motor speed according to the viscosity drop trend, maintaining the fluid shear force in a preset target interval, and avoiding mechanical damage to cells; calculating the mechanical energy dissipation power in real time, and performing feedforward temperature control compensation to eliminate temperature overshoot caused by thermal conduction lag. The application realizes closed-loop control of the physical phase evolution of a heterogeneous tissue dissociation process, solves the limitation of a fixed parameter in a traditional method, shortens the dissociation time, and significantly improves the single-cell yield and survival rate.

This invention relates to the field of battery processing technology, and in particular to a multi-station blade battery cutting and core extraction device, comprising an integral processing frame. A cutting unit, a cross-cutting unit, and a core extraction unit are respectively mounted and connected on the upper surface of the integral processing frame. A material transfer bracket is fixedly mounted on the upper surface of the integral processing frame. Parallel linkage between the units is achieved through a linear slide module. During longitudinal cutting, the material transfer mechanism is synchronously ready and immediately performs the material transfer action after cutting, without any downtime. The material transfer, clamping, cutting, and flipping actions of the cross-cutting unit are continuously connected. The core extraction unit and the cross-cutting unit seamlessly coordinate their posture adjustments. The entire process requires no manual intervention. From battery loading, positioning, and cutting to cell extraction and material diversion, a closed-loop operation is formed. Simultaneously, the timing of each executing component is precisely controllable, effectively shortening the cycle time of a single process, meeting the capacity requirements of large-scale mass production and recycling / dismantling, and significantly reducing labor costs and operational risks.

The invention provides a cell image registration method and system, electronic equipment and a storage medium. The cell image registration method comprises the following steps: acquiring cell contours in a cell image map and a space transcriptome data map; respectively determining a part of target cells based on the definition of the cell contour; extracting geometric centers of a part of target cells, and respectively forming a cell image two-dimensional point cloud set and a space transcriptome data graph two-dimensional point cloud set; and finally obtaining registration parameters. And registering the cell image map and the spatial transcriptome data map according to registration parameters. According to the method, the cell contours and the definition of the cell contours in the cell image map and the space transcriptome data map are obtained, then the two-dimensional point cloud sets are formed based on the geometric centers of part of target cells, point cloud registration algorithm processing is carried out on the two-dimensional point cloud sets, and finally the registration parameters are obtained to carry out registration on the cell image map and the space transcriptome data map. The defect that in-situ analysis cannot be carried out is overcome, and the accuracy of a space transcriptome sequencing technology in a space-time omics technology is improved.

The utility model discloses a processing tank heating and clamping device and a processing tank heating system, and belongs to the technical field of medical instruments, and the processing tank heating and clamping device comprises a fixing mechanism which can fix a processing tank and is provided with a heating unit capable of releasing heat; the heating unit comprises a heating surface which can be attached to the outer wall of the treatment tank; the shaking mechanism is connected with the fixing mechanism and can shake the fixing mechanism, after the adipose tissue is put into the treatment tank, when cells are extracted, the working liquid mixed with a reagent and the adipose tissue can be heated, so that the temperature of the working liquid is guaranteed, and when fat solidification is avoided, the cell extraction efficiency is improved. The activity of cells in the working liquid is ensured; the activity of the biological enzyme in the working liquid is ensured, so that the fat splitting efficiency is ensured.

The utility model relates to the technical field of stem cell filtration, and discloses filtering equipment for stem cell extraction, which comprises a feeding cavity, a filtering cavity and a storage cavity, and further comprises a detection part, a filtering part and a control part, the stirring part is arranged in the center of the top of the feeding cavity, the detection part is located on one side of the stirring part, and the bottom end of the stirring part extends into the feeding cavity; the vibration part is arranged in the filtering cavity in a penetrating mode, and the stirring part makes contact with the top end of the vibration part when rotating to work. By arranging the feeding pipe, the floating ball and other structures, material supplementing can be automatically carried out, the situation that a large amount of solution enters the filtering equipment at the same time is prevented, filtering can be carried out at a stable speed, and by arranging the impacting assembly, the filtering plate can be impacted and vibrated in the stirring process, so that the filtering efficiency is improved. The filtering plate is prevented from being blocked, so that the smooth proceeding of a filtering procedure is ensured, and the stability of the device is improved.

The invention discloses a mouse primary cardiac endothelial cell extraction method, and belongs to the technical field of biological cell extraction. The method comprises the following steps: step 1, obtaining pure myocardial tissues; 2, the treated heart tissue is transferred into an intelligent tissue grinding system to be ground; step 3, transferring the ground heart tissue into a temperature-controlled enzymolysis reactor, automatically monitoring the digestion process, and terminating the reaction when a preset digestion degree is reached; step 4, carrying out primary cell separation, carrying out magnetic separation by using anti-CD31 magnetic beads, and specifically enriching heart endothelial cells; 5, transferring the purified endothelial cells to an intelligent microenvironment culture system, and automatically optimizing a culture medium replacement period to adapt to a cell metabolism state; and step 6, performing multi-dimensional quality evaluation on the endothelial cells, wherein the multi-dimensional quality evaluation comprises marker expression analysis, in-vitro lumen formation experiment, AI-assisted morphological analysis and digital quality scoring.

The invention belongs to the technical field of antibody preparation, and particularly relates to an anti-NKG2D nano antibody as well as a preparation method and application thereof. The method comprises the following steps: constructing and expressing NKG2D protein, immunizing alpaca to obtain immune alpaca PBMC (peripheral blood mononuclear cells), extracting RNA (ribonucleic acid) of the PBMC, carrying out reverse transcription to obtain cDNA (complementary deoxyribonucleic acid), obtaining an antibody gene segment through PCR (polymerase chain reaction), and constructing the antibody gene segment on a phage expression vector; the method comprises the following steps: carrying out electrotransformation on a vector with an antibody gene segment to an SS320 electrotransformation competent state to obtain an NKG2D immune antibody library, carrying out two rounds of NKG2D protein panning and Clone ELISA detection to obtain an antibody combined with NKG2D protein, carrying out sequencing analysis, selecting a unique sequence, and carrying out high-throughput expression through a mammalian cell high-throughput expression system to obtain an antibody protein. And carrying out FACS detection by using an NKG2D high-expression cell line to finally obtain the antibody combined with the NKG2D. The method disclosed by the invention is simple and quick to operate; a mammalian cell expression system induces high-efficiency expression of the antibody, processing modification can be performed after translation, and the activity of the antibody is closer to that of a natural antibody.