20 results about "Tissue digestion" patented technology

The invention discloses a gradient digestion, extraction and detection method for microplastics in plant tissues, and belongs to the technical field of plant pollutant detection. The core of the method is that a hydrogen peroxide-based primary digestion solution, a cellulase and pectinase compounded tissue digestion solution and a residual digestion solution composed of a Fenton reagent and sodium bromide are sequentially adopted to perform three-stage digestion on a plant sample under a mild acidic condition so as to thoroughly remove organic matters and completely retain micro-plastics. After digestion, micro-plastics are extracted through filtration, ultrasonic enrichment and concentration, and finally accurate qualitation and quantification are carried out through thermal cracking-gas chromatography / mass spectrometry. The invention synchronously protects the special digestion solution composition and the kit for realizing the method. According to the scheme, the contradiction between incomplete removal of organic matters and damage of a micro-plastic structure in a traditional digestion technology is effectively solved, the unification of efficient impurity removal (gt, 90%) and high recovery rate (greater than or equal to 85%) is realized, and the accuracy and reliability of detection are remarkably improved.

The utility model relates to the technical field of cell culture, and discloses a cell shake flask capable of improving tissue digestion efficiency, which comprises a culture flask body, a flask cover is mounted at the top end of the culture flask body, and a first constant-temperature heating cover is arranged on one side of the outer side wall of the culture flask body. Mounting blocks are fixed to the front end and the rear end of one side of the first constant-temperature heating cover correspondingly, inner cavities are formed in the first constant-temperature heating cover and the second constant-temperature heating cover correspondingly, and electric heating wires are mounted in the inner cavities. When the culture bottle body disclosed by the utility model is used for culturing cells, the culture bottle body can be heated according to actual conditions by arranging components such as the first constant-temperature heating cover, the mounting block, the mounting groove and the second constant-temperature heating cover, and meanwhile, the digestion efficiency of cell tissues can be promoted by heating the culture bottle body; and the culture bottle body can be detached when not needed subsequently, so that the observation of the condition of cells in the culture bottle body through the culture bottle body is not influenced, and the use effect is better.

The invention discloses a pig jejunum crypt separation and organoid model construction method, and belongs to the technical field of animal tissue separation and culture. The method specifically comprises the following steps: intestinal tract collection, cleaning solution preparation, intestinal tract treatment, tissue trimming, tissue cleaning, digestive juice preparation, tissue digestion, cell mass separation, crypt separation, crypt counting, crypt collection, crypt inoculation and organoid culture. The porcine jejunum organ obtained by the method has a typical three-dimensional cystic structure, has complete epithelial polar and functional cell types, including microvillus, goblet cells, panning cells and endocrine cells, and shows good barrier function and drug metabolic activity. A reliable in-vitro model platform is provided for pig intestinal development, nutrient absorption, pathogen infection mechanism research and precision medical treatment, meanwhile, a technical foundation is laid for large-scale application of organoids, and the method has important scientific significance and clinical value.

The invention discloses a preparation method of hydrolase suitable for space transcriptome poplar capsule tissue digestion. The preparation method comprises two steps of reagent preparation and experiment. A digestion buffer solution and a washing buffer solution need to be prepared for reagent preparation, and the experimental steps comprise setting of instrument temperature, preparation of various solutions, and treatment of tissue slices, such as fixation, dyeing, cleaning, incubation and the like. The tissue digestion hydrolase solution contains various enzymes such as pectinase Y-23 and xyloglucanase and a sodium citrate buffer solution. The method provides technical support for the poplar capsule tissue in spatial transcriptome research, adapts to digestion of the poplar capsule tissue, adopts common enzymes, reduces cost, has low technical requirements on experimenters, has a good digestion effect and a clear cell structure, provides a high-quality sample for subsequent research, and is beneficial to revealing molecular characteristics of the poplar capsule tissue.

The utility model provides a novel tissue digester which comprises a digester shell and a sealing cover, a first chamber partition plate and a second chamber partition plate are installed in the digester shell, a tissue digestion chamber is formed between the first chamber partition plate and the second chamber partition plate, and a tissue digestion liquid channel is installed on the left side of the second chamber partition plate in a communicated mode. The left side of the first chamber partition plate is communicated with a cell suspension channel with a baffle, the interior of the first chamber partition plate is communicated with a filter screen, the filter screen is aligned with an input port of the cell suspension channel with the baffle, two sealing strips are installed below the sealing cover, and cover buckles are installed on the front side, the rear side, the left side and the right side of the sealing cover respectively. According to the method, digestion of released cells can be terminated, the activity of the cells is kept, tissue digestion is not affected, the cells can be continuously released, the effective digestion time is far longer than t, and the number and quality of the finally obtained cells are greatly improved.

The invention relates to the technical field of immune cells, in particular to a preparation method of human placenta-derived NK cells. The preparation method comprises the following steps: S1, separating placenta tissues; s2, placenta tissue digestion; S3, mononuclear cell separation; S4, NK cell sorting; and 5, NK cell multiplication culture.

The present application relates to the technical field of biology, in particular to buffalo mammary gland tissue digestion solution and application thereof, and provides a preparation method of buffalo mammary gland tissue digestion solution and buffalo mammary gland tissue single-cell suspension, and the preparation method can obtain a primary cell suspension of buffalo mammary gland tissue with high viability, low clumping rate, high nucleated rate and less impurities, and can greatly improve the success rate of single-cell sequencing experiment of buffalo mammary gland tissue.

The present application relates to the technical field of biology, in particular to human thymus tissue digestion solution and application thereof. The present application provides a preparation method of human thymus tissue digestion solution and human thymus tissue single cells, and the preparation method can obtain a human thymus tissue primary cell suspension with high viability, low fine nodule rate, high nucleated rate and less impurities, and can greatly improve the success rate of single cell sequencing experiment of human thymus tissue primary cells.

The invention discloses an efficient universal mouse pancreatic tissue single-cell suspension preparation method, and belongs to the technical field of cell biology. Pancreas are rich in digestive enzymes, so that the treatment difficulty of in-vitro cells is high, and a high-quality single-cell suspension is difficult to prepare by a traditional method. The method comprises the steps of reagent preparation, tissue sampling, tissue dissociation, red cracking and cleaning, AOPI dyeing quality inspection and the like. Components and concentrations of a tissue enzymatic hydrolysate, a cell cleaning solution and a cell suspension are determined in reagent preparation; the acquisition method of the pancreatic tissue of the mouse is defined by tissue sampling; the tissue dissociation adopts a two-round digestion mode, and the cut tissue is subjected to enzymolysis by using a compound enzyme solution under specific conditions; the cell precipitates are treated through red cracking and cleaning; aOPI dyeing quality inspection is used for analyzing cell count and motility rate. According to the method, the culture medium containing specific components and the compound enzyme liquid are creatively used, the problem that pancreas single cells are difficult to dissociate can be effectively solved, the single-cell suspension with high motility rate, high purity and low adhesion is obtained, and the requirements of downstream single-cell experiments are met.

The present application relates to the field of biotechnology, in particular to human bladder tissue digestion solution and application thereof.The digestion method of the present application can dissociate human bladder tissue cells while keeping the cell viability at more than 90%, and the amount of harvested cells is also obviously improved.Sufficient cell amount is the prerequisite for optimization treatment such as red blood cell lysis and dead cell removal.The single cell suspension obtained by the method has high viability, low clumping rate, high nucleated rate and less impurities, which can provide a basis for obtaining high-quality gene expression data in human bladder tissue single cell sequencing experiment.The present application can quickly obtain human bladder tissue primary cell suspension (sufficient cell amount, high viability, low clumping rate and less impurities) meeting the requirements of single cell experiment within 2-4 hours.

The invention discloses an ultrasonic-assisted digestion device for hair follicle cells, which comprises a shell, a tissue digestion tank, a cell collection tank and a pipeline are arranged in the shell, one end of the pipeline is communicated with the tissue digestion tank, the other end of the pipeline is communicated with the cell collection tank, and a tissue transfer cylinder is mounted on the tissue digestion tank. The tissue transfer cylinder is communicated with the upper surface of the shell, a cell collection cylinder is arranged on the cell collection tank and is communicated with the upper surface of the shell, an ultrasonic generator is arranged on the outer wall of the tissue digestion tank, and an ultrasonic switch electrically connected with the ultrasonic generator is arranged on the upper surface of the shell; the invention further discloses a using method of the device, the device combines an ultrasonic technology and a chemical digestion method, hair follicle cells can be efficiently and mildly separated from hair follicle tissue, cell damage is effectively avoided, and the cell recovery rate is increased; the device is suitable for rapid separation and culture of hair follicle cells.

The invention discloses a lung cancer vascularized organ and application thereof in mouse tumor formation model and drug screening, and belongs to the technical field of biomedicine and drug research and development. A preparation method of a lung cancer vascularized organoid comprises the following steps: taking a lung cancer tissue or a needle biopsy sample, and treating with tissue digestive juice to obtain single cells or tissue fragments; embedding the single cells or tissue fragments obtained in the step 1 with matrigel, and placing the single cells or tissue fragments in a special lung cancer culture medium containing EGF, FGF and Wnt growth factors to form lung cancer organs; and adding two pro-vascularization factors, namely VEGF and Angiopoietin-2, into the lung cancer organoid culture system in the step 2, so as to obtain the lung cancer vascularization organoid. The method solves the problems that traditional organoid lacks a functional vascular network and is insufficient in physiological correlation, provides a more accurate research tool for lung cancer disease modeling and drug screening, and can be widely applied to the fields of accurate medicine, oncology research and new drug research and development.

The invention relates to the technical field of neural stem cell tests, in particular to a preparation method of a mouse midbrain tissue neural stem cell high-activity single-cell suspension. The method comprises: obtaining a mouse embryo; treating the mouse midbrain tissue to obtain a cell digestion suspension of the mouse midbrain tissue; filtering and centrifuging to obtain a mouse midbrain single-cell suspension, and performing culture and passage by using a neural stem cell culture medium to obtain P1-generation or P2-generation mouse midbrain neural spheres; resuspending the P1-generation or P2-generation mouse midbrain nerve spheres by using a culture medium, digesting the P1-generation or P2-generation mouse midbrain nerve spheres by using or not using tissue digestive enzymes, and culturing by using the following culture medium to obtain the single-cell suspension of the mouse midbrain tissue neural stem cells. According to the application, the survival rate of the single cells of the neural stem cells is ensured to be more than 95%, the cell library building requirement is met, and efficient living cells are provided for Single-cell sequencing, ATCT-Sequence sequencing and the like of the neural stem cells.

The utility model relates to a kind of filtering devices for amniotic membrane tissue digestion cell liquid, belong to the technical field of cell liquid processing equipment.It is formed by the cooperation of first filter assembly, second filter assembly and filter cup assembly, and forms sealed flow channel in vertical direction, accelerates high viscous liquid flow in the whole filtration process, reduces blockage;At the same time, the flow guide pipe is connected to the lowest point of filter cup, and cooperates with liquid stop clamp to realize rapid collection, reduce sample loss;Single operation completes filtration operation, eliminates external pollution and eliminates the risk of liquid leakage.

The invention discloses a mouse intestinal tract organ and a culture method and application thereof. The method comprises the following steps: extracting fetal mouse colon tissues from pregnant mice, treating the fetal mouse colon tissues with digestive enzymes to prepare a cell mass suspension, and further mixing the cell mass suspension with a matrigel seed plate for culture. According to the method, the forming efficiency and success rate of the colon-like organs are remarkably improved, and the method has wide application prospects in the fields of disease modeling, drug screening and the like.

The present application relates to the field of cell detection and screening, and particularly relates to a method and system for detection and screening of amniotic epithelial stem cells. The method comprises: generating a structured sample task sheet based on multi-source information; performing tissue digestion processing based on the task sheet to obtain a candidate cell group; then performing phenotype detection, molecular detection and quality detection to generate corresponding data packets and label data packets; based on the quality label data packets, performing gating, batch normalization, abnormality rejection, fusion scoring and consistency determination processing through a screening engine to generate a screening result packet; and finally generating a disposal work order and a traceability report based on the screening result packet. The present application realizes standardized and auditable automatic processing of the whole process of amniotic epithelial stem cells from receiving, detection to sorting and disposal, greatly improving the detection efficiency, result consistency and traceability.

The invention relates to the technical field of cell culture, in particular to a preparation method and kit for an organoid culture medium, and the preparation method for the organoid culture medium comprises the following steps: sequentially mixing serum-free B27, nicotinamide and other components to obtain a first solution; adding Wnt3a, RSPO1 and Noggin with corresponding concentrations on the basis of whether the organoid has KRAS mutation or not to obtain a second solution; selectively adding an N-2 additive or estradiol based on the type of the tumor organoid to obtain a third solution; and finally, DMEM (Dulbecco Modified Eagle Medium) based and Advanced DMEM / F12 are added to obtain the organoid culture medium. According to the invention, the concentration and combination of key components in the culture medium can be dynamically adjusted according to three key characteristics of the type of the tumor organoid, whether the tumor organoid is a KRAS mutant type or not and the content of tumor parenchymal cells. The kit comprises an organoid culture medium, a rinsing solution, a tissue digestion solution, a neutralization solution and a passage digestion solution. The defect that an existing fixed formula culture medium is difficult to adapt to sample heterogeneity is overcome, and the success rate and culture stability of organoid construction are improved.

The invention relates to a culture method of bile duct cancer organoid. The culture method comprises the following steps: S1, cleaning bile duct cancer tissues in a biosafety cabinet, and chopping the bile duct cancer tissues; s2, transferring the chopped tissue into a centrifugal tube filled with tissue digestive juice for digestion, and collecting an upper suspension; s3, centrifuging the collected upper suspension, and collecting bile duct cancer tissue cell precipitate; s4, inoculating and embedding bile duct cancer tissue cells into matrigel; s5, after the matrigel is solidified, culturing the tissue cells by using a corresponding organoid culture medium, and obtaining primary organoid; the culture medium is replaced every three days. Due to the adoption of the technical scheme, the culture method of the bile duct cancer organoid can be used for efficiently and stably culturing the bile duct cancer organoid.

This invention relates to the field of in vitro intestinal organoid culture technology, and discloses an in vitro method for constructing a small intestinal organoid model of the Chinese red panda. The method involves collecting ileum tissue from Chinese red pandas that have died due to disease or accident, and then using PBS... + After rinsing, the tissue was placed in tissue transport buffer for low-temperature transport; the following steps were performed sequentially: intestinal lumen rinsing, removal of external tissue, longitudinal incision, scraping of intestinal villi onto a glass slide, and pre-cooling with PBS. + The tissue was repeatedly inverted and washed until the supernatant was clear. It was then minced, digested with EDTA digestion solution, and the crypts were separated by mechanical pipetting. After filtration through a sieve, the crypt precipitate was collected by centrifugation. The crypt precipitate was resuspended, its concentration adjusted, and mixed with Matrigel. It was then inoculated into culture plates, and after the Matrigel solidified, complete culture medium was added for further culture to obtain small intestinal organoids from the Chinese red panda. A small intestinal organoid model was successfully constructed and can be stably passaged in vitro, filling a gap in the technology for constructing small intestinal organoids from the Chinese red panda.

The present application relates to the technical field of biology, in particular to mouse common carotid artery tissue digestion solution and application thereof, and provides a preparation method of mouse common carotid artery tissue digestion solution and mouse common carotid artery tissue single cell. The preparation method can obtain a mouse common carotid artery tissue primary cell suspension with high viability, low clumping rate, high nucleated rate and less impurities, and can greatly improve the success rate of mouse common carotid artery tissue single cell sequencing experiment.