965results about "Vertebrate cells" patented technology

The invention provides a pancreatic islet organ capable of efficiently maintaining pancreatic islet beta cell identity and a preparation method and application thereof. Specifically, the invention provides the preparation method for the in-vitro pancreatic islet organ, and the preparation method comprises the following steps: carrying out induced differentiation culture on the ZnT8-knocked-out pluripotent stem cells, so as to obtain the mature pancreatic islet organ. The invention also provides a corresponding transplantation composition and application thereof. The invention discloses that ZnT8 knockout is a key factor for effectively maintaining the identity stability of beta cells in pancreatic islet organs. In addition, the pancreas islet beta cell identity maintenance agent is combined with the ZnT8-knocked-out pancreas islet organ for use, so that the identity stability of the pancreas islet organ beta cell can be obviously maintained. The invention provides a new strategy for transplantation treatment of type 2 diabetes, and has excellent clinical transformation value.

The invention belongs to the technical field of biology, and particularly relates to a high-throughput preparation method of a single-cell-derived organoid based on a microcapsule spherical shell structure, and the preparation method comprises the following steps: preparing an aqueous two-phase microcapsule encapsulated with a single cell, culturing the single cell in the aqueous two-phase microcapsule to obtain the organoid, the substances of the inner phase and / or the outer phase respectively comprise any one or a combination of more of the following substances: methacrylated gelatin, carboxymethyl cellulose, glucan, sodium alginate and polyvinyl alcohol. The invention also provides the organoid prepared by the preparation method and application of the preparation method. According to the invention, the microcapsule with a permeable core-shell structure is formed by packaging the microcapsule in a single-cell form in two-aqueous-phase liquid drops, the wrapping rate is effectively controlled and adjusted on the premise of conforming to Poisson distribution by optimizing parameters, and the single-cell wrapping rate and the organoid single-cell induction efficiency are optimized.

The invention belongs to the field of biotechnology and aquaculture, and particularly relates to a method for improving hypoxia resistance of pelteobagrus vachelli based on hypoxia induction of liver extracellular vesicles of the pelteobagrus vachelli. The method comprises the following steps: extracting high-purity extracellular vesicles EVs in liver tissues of pelteobagrus vachelli subjected to low-oxygen treatment by adopting a method of combining differential centrifugation and high-precision iodixanol density gradient centrifugation; diluting the extracted EVs to 2mu g / mu L by using PBS (Phosphate Buffer Solution) to prepare an EVs suspension; an intraperitoneal injection mode is adopted, the EVs suspension is injected into the body of the pelteobagrus vachelli, and after injection is conducted for 24 hours, the hypoxia resistance of the pelteobagrus vachelli is evaluated through a suffocation point detection test. The invention opens up a brand new path for improving the hypoxia resistance of the pelteobagrus vachelli, provides a new idea and a new method for efficiently enhancing the hypoxia resistance of economic fishes in the field of aquaculture, is expected to play an important role in actual aquaculture production, and assists the sustainable development of the aquaculture industry.

The invention relates to a culture medium for high-density serum-free suspension culture of BHK-21 cells and application of the culture medium. The culture medium contains inorganic salt, amino acid, vitamins, energy substances, a buffer substance, an antioxidant, sterol, polyamine, an additive and an indicator, and the antioxidant comprises dihydrolipoic acid and S-acetyl-L-glutathione. According to the invention, the culture medium for serum-free suspension culture of the BHK-21 cells, which is clear in components and remarkable in culture effect, can be provided. The added dihydrolipoic acid and S-acetyl-L-glutathione are combined as antioxidants of the culture medium, so that the survival rate of BHK-21 cells cultured in the culture medium is improved while the oxidation resistance of the serum-free suspension culture medium is improved.

The invention relates to a method for extracting high-purity extracellular vesicles from liver tissues of pelteobagrus vachelli, which comprises the following steps of: firstly, mixing collagenase II and collagenase IV, performing enzymolysis on the tissues, adding a CaCl2 solution, incubating, and digesting the liver tissues into a tissue suspension; then removing cells and fragments through low-speed centrifugation, and filtering to remove impurities by using needle filters of 0.8 [mu] m and 0.22 [mu] m; and finally, further purifying by combining ultracentrifugation with a high-precision iodixanol density gradient centrifugation method, and merging density layers with high purity to obtain high-purity EVs. The method has the characteristics of high extraction purity, high efficiency and simplicity and convenience in operation, and has a wide application prospect.

The invention belongs to the technical field of cell biology, and particularly discloses a culture medium and a culture method of fish primary liver cells. The culture medium of the fish primary liver cells comprises the following components: 90% of an M199 culture medium, 10% of fetal calf serum, 100 U / mL of penicillin, 0.1 mg / mL of streptomycin, 5 ng / mL of EGF, 10 [mu] U / ml of insulin and 10 mM of dexamethasone. Based on the culture medium, an efficient and stable fish liver cell culture method is developed by combining a standardized anesthesia and disinfection process, a precise dissection and liver sampling method, an optimized tissue digestion and cell separation scheme, an efficient cell purification and adherent culture system, a stable passage technology and the like; a standardized hepatocyte line establishment operation process is defined, and the problems that species binding exists in an existing fish liver cell culture method, and a standardized culture process suitable for multiple fish fingerlings is difficult to form are solved.

The invention discloses a cat kidney suspension culture cell line suitable for serum-free suspension culture and application of the cat kidney suspension culture cell line. The name of the cell line is F81-B4, and the preservation number is CGMCC (China General Microbiological Culture Collection Center) No.46350. After 50 generations of continuous subculture, the cells still can maintain the original proliferation level and cellular morphology, the cell number can reach 9.0 * 10 < 6 > / mL after the cells are subcultured at the density of 1.0 * 10 < 6 > / mL for three days, the multiplication time is 22-26 hours, and the cell proliferation speed is equivalent to that in a shake flask when the cells are amplified to a 10L bioreactor for culture. The cell line is highly susceptible to feline parvovirus, feline calicivirus, feline herpes virus, canine parvovirus and canine distemper virus, the virus content of a virus solution harvested 72 h after FPV virus inoculation can reach 107.5 TCID50 / mL, the virus content of a virus solution harvested 24 h after FCV virus inoculation can reach 1010.8 TCID50 / mL, the virus content of a virus solution harvested 48 h after FHV-1 virus inoculation can reach 107.5 TCID50 / mL, compared with an existing F81 adherent cell production process, the production process has the advantages that the production efficiency is high, and the production cost is low. The method has the characteristics of rapid lesion, short virus collection time and high titer. The cell matrix is an ideal cell matrix which can be used for culturing the three viruses.

The invention discloses a combined evaluation method for in-vitro and in-vivo red fading and relieving effects of cosmetic raw materials, which comprises the following steps: in-vitro model evaluation: inducing a human immortalized umbilical vein endothelial cell model by adopting TNF-alpha, and detecting the secretion amount of a prostacyclin I2 factor to obtain a score of an in-vitro model; in-vivo model evaluation: adopting a chick embryo chorioallantoic membrane model, and obtaining a score of the in-vivo model according to the degree of vasoconstriction caused by the to-be-detected raw material sample acting on the chick embryo chorioallantoic membrane; and comprehensive evaluation: based on the score of the in-vitro model and the score of the in-vivo model, comprehensively evaluating the red fading and relieving efficacy of the raw material sample to be tested. According to the technical scheme, the defect that in the prior art, no cosmetic raw material soothing efficacy evaluation method for the vascular reactivity increasing mechanism exists is overcome, efficacy evaluation is more accurate, the method has the advantages of being comprehensive, short in test period, easy to operate, economical and convenient, and an effective reference basis is provided for screening and compounding schemes of red fading and soothing raw materials.

The invention discloses a hydrogel microsphere sorting method, an artificial organ preparation method, a system and a medium. The method comprises the following steps: acquiring a hydrogel microsphere image in the micro-fluidic chip; analyzing the hydrogel microsphere image through a neural network to obtain an analysis result; based on the analysis result, hydrogel microspheres wrapping the target cells are sorted out; the neural network comprises a feature extraction layer and a multi-scale joint output head, and the hydrogel microsphere image is processed by the multi-scale joint output head after being convolved by the feature extraction layer to obtain an analysis result. According to the method, the hydrogel microspheres wrapped with the target cells are accurately identified and sorted by using a droplet microfluidic technology, a standardized artificial organ can be prepared based on the sorted hydrogel microspheres, and the functional maturity and reliability of the artificial organ are remarkably improved.

A microfluidic 3D-printed hydrogel based on fish liver decellularized extracellular matrix for liver regeneration and a preparation method. The hydrogel is prepared by means of combining fish liver decellularized extracellular matrix (dECM) and gelatin methacryloyl (GelMA), and loading induced pluripotent stem cell-derived hepatocytes (iPSC-heps) for liver regeneration. The microfluidic 3D-printed hydrogel based on fish liver decellularized extracellular matrix exhibits excellent biocompatibility, retains intact endogenous growth factors, maintains the biological activity of cells, ensures the effective encapsulation of the cells, and facilitates the robust functional expression of the iPSC-heps. After in-vivo transplantation, the survival rate and liver function of mice with acute liver failure are significantly improved, and liver regeneration and repair are promoted.

The invention discloses a preparation method of vascular tissue-derived extracellular vesicles, and belongs to the field of cytobiology. According to the preparation method of the extracellular vesicles, the artery blood vessel tissue is directly subjected to enzymolysis through collagenase, the extracellular vesicles are obtained by combining low-speed centrifugation and ultra-speed centrifugation methods and utilizing a differential centrifugation mode, and the technical blank of obtaining the extracellular vesicles from the blood vessel tissue is filled. Experimental results show that the extracellular vesicles can be successfully prepared from collagenase I, collagenase II, collagenase IV, collagenase V or collagenase IV combined with elastin, the prepared extracellular vesicles are different in particle size, form, yield, protein quantity, purity and the like, and the quality of the extracellular vesicles obtained by enzymolysis of collagenase IV combined with elastin is optimal. The extracellular vesicles are extracted from vascular tissues with normal physiological characteristics for the first time, a new source is provided for preparation of the extracellular vesicles, and the extracellular vesicles have great significance in expanding application of the extracellular vesicles.

The application discloses a construction method and application of an ovary cell line of Quasipaa spinosa. The method comprises the following steps: selecting metamorphosis period larva ovary tissue, disinfecting and obtaining the ovary tissue, and then using collagenase I digestion solution to digest the ovary tissue at 26 DEG C for 1 hour; after the tissue block adheres to the wall, using a low-osmotic-pressure complete culture medium to carry out primary culture at 25-27 DEG C, wherein the culture medium is 60% diluted DMEM / F12 culture medium containing 10%-15% fetal bovine serum; when the cell confluence degree is higher than 70%, using trypsin-EDTA to digest the cells for 1-2 minutes to subculture, and the subculture culture medium contains 30% cell conditional culture medium; and finally, the cells are cryopreserved through programmed cooling. The ovary epithelial cell line of Quasipaa spinosa is successfully established for the first time, the survival rate of the cells after recovery is high, and the karyotype is stable. The constructed cell line can be used for amphibian reproductive biology research, compound screening and in-vitro conservation of germplasm resources.

Disclosed herein are cell processing systems, devices, and methods. The system for cell processing can include multiple instruments, each independently configured to perform one or more cell processing operations on a cartridge, and a robot capable of moving the cartridge between each of the multiple instruments.

The invention provides a composition containing exosome, which is prepared from the following components in parts by weight: 10 to 20 parts of exosome, 5 to 15 parts of extract, 0.3 to 0.5 part of antioxidant and 0.05 to 0.1 part of preservative, the extract is a mixture of 3-8 parts of aloe extract, 1-5 parts of camellia flower extract and 1-3 parts of cranberry extract; the preservative is one or more of benzoic acid, salicylic acid, boric acid and sorbic acid; the antioxidant is propyl gallate. When the composition is applied to an anti-aging product, the temperature sensing effect and the DPPH free radical scavenging rate are obviously improved.

This application relates to the field of cell culture medium production technology, specifically disclosing an improved method for producing lyophilized cell culture medium powder. The method includes the following steps: S1, grouping the trace elements according to their mass percentage in the cell culture medium formulation, solubility, and pH level after dissolution; S2, mixing each group of trace elements with a lyophilization carrier in purified water to prepare a homogeneous lyophilized solution; S3, subjecting the lyophilized solution obtained in S2 to vacuum freeze-drying to obtain lyophilized trace element powder; S4, mixing the lyophilized trace element powder obtained in S3 with other components of the cell culture medium to obtain the finished cell culture medium powder. The lyophilized powder production method of this application can be used for the efficient preparation of lyophilized cell culture medium powder. It has the advantages of ensuring the solubility and chemical stability of the lyophilized solution components, ultimately obtaining a lyophilized powder with uniform components and good solubility, avoiding precipitation and detection deviations, and improving the batch stability of the finished culture medium.

The application discloses a device for constructing an open vascularized organ chip, comprising a substrate layer, a channel layer and a culture medium cavity layer; the channel layer has at least one channel unit; the channel unit has a first channel and two or more second channels, the second channels are provided with culture medium inlets and outlets, and the second channels have a preset height difference with the first channel; the culture medium cavity layer has a plurality of culture medium cavities; the substrate layer, the channel layer and the culture medium cavity layer are designed in a split mode, and when stacked from bottom to top, the culture medium cavities correspond to and communicate with the culture medium inlets and outlets one by one. Further, the application also discloses a method for constructing an open vascularized organ chip by using the device and an obtained open vascularized organ chip. The culture medium cavity layer and the channel layer of the organ chip are designed in a split mode, and based on this, high flow rate gap flow through the interior of the matrix gel and pipeline flow along the side wall of the matrix gel are realized in the chip.

The present disclosure describes compositions, preparations, nanoparticles (such as lipid nanoparticles), and / or nanomaterials, and methods of using the same.

The present invention provides a method for safely and simply performing treatments such as exfoliation / floating treatment, dormancy treatment (dormancy not only enhances the protective effect but also suppresses unnecessary differentiation of cells, thereby maintaining an undifferentiated / low-differentiated state), and protective treatment for storage and transportation, on cells in tissue or cultured cells, while preventing their death and maintaining their viability, and a method for safely and simply performing cell activation treatment for implanting dormant floating cells. The cell treatment agent contains, as an active ingredient, at least one selected from alginic acid, heparins, sulfated dextran, and protease inhibitors.

To provide a method for creating a tissue construct including two or more individual regions each of which includes a different cell population or cell line.SOLUTION: A method includes: a step of providing a support base material or a substrate assembly including two or more physically separate regions, in which two or more physically separate regions of the support base material or the substrate assembly are different from each other; and a step of depositing / arranging one or more cells on the support base material or the substrate assembly. The cells have at least two zones (e.g. proliferation zone and non-proliferation zone), and can form a continuous monolayer capable of operating as an in-vitro intestinal model. The model is two-dimensional, and therefore, facilitates quick and light imaging. A system including a tissue construct, and a method for measuring the effects of a medicine, a nutritional supplementary food and a metabolite for enterocytes by using a tissue construct are also provided.SELECTED DRAWING: Figure 1C

Targeted integration (TI) host cells suitable for the expression of recombinant proteins, and methods for making and using the TI host cells, are provided. [Solution] The TI host cell contains an exogenous nucleotide sequence integrated into an integration site within a specific locus in the genome of the host cell, and the nucleotide sequence immediately 5' to the integrated exogenous nucleotide sequence consists of a sequence that is at least 90% homologous to nucleotides 69303-79768 of NW_003616412.1.

The invention discloses a porcine circovirus type 2 and haemophilus parasuis bivalent inactivated vaccine and a preparation method thereof. According to the invention, a porcine circovirus strain, a haemophilus parasuis serum type 4 strain and a haemophilus parasuis serum type 5 strain which are epidemic in situ are respectively separated and obtained, and the microbial preservation numbers of the porcine circovirus strain, the haemophilus parasuis serum type 4 strain and the haemophilus parasuis serum type 5 strain are respectively CGMCC (China General Microbiological Culture Collection Center) No.47094, CGMCC No.46698 and The separated strain or bacterial strain is prepared into a bivalent inactivated vaccine, and lentinan is added in the vaccine preparation process to enhance the immune effect; a safety evaluation result shows that local or whole-body adverse reaction caused by the bivalent vaccine does not occur, so that the safety of the bivalent vaccine is good; the immune efficacy evaluation result of the bivalent vaccine shows that the bivalent vaccine can provide complete protection for animals aiming at the in-situ epidemic porcine circovirus, haemophilus parasuis serotype 4 and haemophilus parasuis serotype 5, and has good immune protection efficacy.

The present invention relates to surface-functionalized extracellular vehicles. Specifically, the present invention relates to exosomes surface-functionalized with lactosylated polyetheneimine (PEI-LA), such surface-functionalized exosomes as drug delivery vehicles, 5 compositions comprising a therapeutic agent encapsulated within such exosomes, methods of producing such exosomes and compositions thereof, as well as methods of delivering such exosomes and compositions to liver.

The present application relates to the application of nucleic acid molecules in the preparation of drugs for treating liver diseases; the nucleic acid molecules have one or more of the following conditions: comprising any one or more of miR-302s; comprising any one or more of miR-302s, mimics or derivatives of any one or more of miR-302s, the mimics or derivatives have a core fragment shown in SEQ ID No. 1 and have the same or similar function as any one or more of the miR-302s members; comprising a miRNA precursor, which can be processed into any one or more of miR-302s, mimics or derivatives in the host; comprising a polynucleotide fragment, which can be transcribed into the miRNA precursor in the host; comprising an expression vector containing a polynucleotide fragment. The present application provides a new idea for the treatment of liver diseases.

The invention relates to viral vector production. In particular, the present invention provides novel methods for producing viral vectors. The invention also provides a corresponding virus vector production system and application.

The present invention relates to a method for producing a stable hydrogel-cell composition comprising the steps of:a) providing a reaction mixture comprising at least one polymerizable substrate and at least one enzyme capable of polymerizing said at least one substrate, and cells, andb) incubating the mixture of step a) to form a stable hydrogel-cell composition.

To provide biological fluid purification using a biocompatible membrane.SOLUTION: An apparatus for integrated adaptive biological blood purification includes a function unit. The function unit is provided with: a membrane including a pipe surface and a filtration surface; a blood vessel channel system which is adhered to the blood vessel surface of the membrane and includes a first lumen space in fluid communication with the blood vessel surface of the membrane, a first end configured to be in fluid communication with a fluid supply part and to contact the same, and a second end configured to be in fluid communication with a filtration fluid outlet and to contact the same; and a filtration channel system which is adhered to the filtration surface of the membrane and includes a second lumen space in fluid communication with the filtration surface of the membrane and a third end configured to be in fluid communication with a filtrate outlet and to contact the same.SELECTED DRAWING: Figure 11

Provided is an RNA plasmid delivery system and its use. The RNA plasmid delivery system includes a plasmid carrying an RNA fragment to be delivered, which is enriched in a host's organ tissue, can endogenously and spontaneously form exosomes containing the RNA fragment in the host's organ tissue, and can enter and bind to a target tissue to deliver the RNA fragment to the target tissue. The RNA delivery system is safe, reliable, has good drug development potential, and is highly versatile.

The invention discloses a non-tumorigenic MDCK cell strain, a screening method and application. The non-tumorigenic MDCK cell strain is MDCK-CA027 (the preservation number is CCTCC (China Center for Type Culture Collection) NO: C2023329). The tumor formation rate of the cell strain is 0% when the cell strain is observed for at least 16 weeks in a nude mouse in-vivo tumor formation test, and the cell strain has good sensitivity to various influenza viruses (such as H1N1, H3N2, BY and BV). The invention also provides a method for screening the cell strain. The method comprises the following steps: preparing a single-cell suspension, carrying out limited dilution to obtain monoclone, carrying out enlarged culture, carrying out virus sensitivity detection, carrying out in-vitro tumorigenicity related detection (clone formation test / soft agar test), carrying out nude mouse in-vivo tumorigenicity test and the like. The non-tumorigenic MDCK cell strain is high in safety, can efficiently support influenza virus replication, can be safely and effectively applied to influenza vaccine production and influenza virus amplification, overcomes the tumorigenic risk of traditional MDCK cells, and has important scientific significance and application prospects.

Provided is a method of intermittent perfusion fed-batch culture, comprising a fed-batch process including one or more intermittent perfusion phases during the middle to late stage to improve productivity and product quality.