232results about "Invertebrate cells" patented technology

The invention relates to application of sea cucumber-derived nano-vesicles in preparation of a medicine for promoting wound repair. The nano-vesicles (SC-EVs) extracted and prepared from sea cucumbers can effectively promote cell migration, scratch repair and tube forming capabilities of human skin epithelial cells (HACAT) and human skin microvascular endothelial cells (HM DEC) and inhibit cell apoptosis, so that the capability of repairing cells is achieved, and particularly the effect of repairing wound surfaces in a high glucose state or diabetes mellitus is achieved. The invention provides a new direction and thought for the application of the sea cucumber-sourced nano vesicles (SC-EVs), provides a brand new drug research and development and clinical treatment direction for wound repair, especially for solving the problem of difficult healing of wounds of patients with high glucose or diabetes, and has extremely high social value and market application prospect.

The invention belongs to the field of biomedicine, and particularly discloses a herpes zoster vaccine as well as a preparation method and application thereof. The invention provides a herpes zoster vaccine. A fusion protein in the herpes zoster vaccine comprises a VZV-gE protein, an Fc and an optional antigen epitope with a cellular immune activation effect. The adjuvant can be a combination of an MF59 adjuvant and a CpG1009 adjuvant, and enhances humoral immunity and cellular immunity at the same time. The vaccine formed by the fusion protein in cooperation with an innovative adjuvant system can induce generation of strong cellular immunity and humoral immunity.

The invention discloses a preparation method and application of a sea cucumber exosome with an immune cell repairing function, and belongs to the technical field of biomedicine. The method comprises the following steps: feeding sea cucumbers by using a combined method of composite stress and an echinacea extract, extracting sea cucumber cells, and obtaining sea cucumber exosomes through ultracentrifugation; wherein the condition of composite stress is set as follows: the day and night temperature difference is not lower than 3 DEG C, and meanwhile, the dissolved oxygen content of the water body is adjusted to 75-90% of the dissolved oxygen content of the conventional feeding water body. The obtained sea cucumber exosome has a remarkable immune cell repairing function, and can effectively promote proliferation and activity recovery of immune cells, inhibit expression of inflammatory factors and improve the level of anti-inflammatory factors. The sea cucumber exosome prepared by the preparation method disclosed by the invention has an important application prospect in the research and development of immune repair drugs, and can provide a technical support for a novel immune regulation preparation in the field of biomedicine.

The invention discloses a haemonchus contortus subunit vaccine, and belongs to the field of veterinary vaccines. The subunit vaccine comprises an antigen protein MEP3, an antigen protein AP1, an antigen protein AP8, an antigen protein H11-2, an antigen protein H11-4 and an antigen protein AP5 of galactosylated and modified haemonchus contortus, and the antigen protein MEP3, the antigen protein AP1, the antigen protein AP8, the antigen protein H11-2, the antigen protein H11-4 and the antigen protein AP5 of galactosylated and modified haemonchus contortus. The galactosylation modification of the antigen protein is realized by jointly expressing glycosyl transferase HcGALT of haemonchus contortus and one or more of the proteins in eukaryotic cells. The antigen protein modified by galactosylation in the subunit vaccine is closer to the glycosylation modification of natural protein, and can provide effective immune protection after the goat is infected with haemonchus contortus, and the egg laying rate is reduced by 81.43%. The invention lays a foundation for development of haemonchus contortus subunit vaccines, provides an effective technical means for prevention and treatment of the haemonchus contortus subunit vaccines, and has important application value and popularization prospect.

The invention relates to the technical field of cell culture media, in particular to a marine invertebrate cell culture medium. Comprising the following components in parts by weight: 0.5 to 1 part of a compound supplement, 1 to 4 parts of glucose, 0.3 to 0.5 part of L-glutamine, 0.01 to 0.05 part of vitamin, 10 to 15 parts of fetal calf serum, 0.02 to 0.04 part of penicillin, 0.01 to 0.03 part of trehalose, 0.3 to 0.7 part of a sodium chloride osmotic pressure regulator, 0.5 to 1 part of inorganic salt and 10 to 25 parts of a sodium bicarbonate buffer agent, wherein the compound supplement is prepared by mixing meson, diatom and disodium cyclohexane diamine tetraacetate according to the mass ratio of 1: (0.2-0.6): (0.1-0.5), the application range of the culture medium is widened by adopting the compound supplement, the meson and the diatom are added into the cell culture medium after enzymolysis, and compared with a chemically synthesized culture medium, the culture medium better meets the requirements of marine invertebrate cells; therefore, cell growth is effectively promoted, and the purposes of rapid growth and stable passage are achieved.

The invention belongs to the field of biomedicine, and particularly discloses a nasal spray influenza vaccine as well as a preparation method and application thereof. The invention provides a nasal spray influenza vaccine. Fusion protein in the nasal spray influenza vaccine comprises influenza virus strain HA protein or fragments thereof, influenza virus protein epitopes and ferritin. The adjuvant may be a CpG1009 adjuvant to enhance humoral and cellular immunity. A vaccine composed of the fusion protein and a novel CpG adjuvant can induce generation of a strong challenge protection effect.

The present invention relates to RSV F protein mutants and their applications, belonging to the field of vaccines. Based on the amino acid sequence of wild-type RSV F protein, the present invention has carried out modifications in multiple aspects and in various ways, including disulfide bond mutations, proline mutations, cavity filling mutations, electrostatic mutations, p27 sequence modifications, and / or trimerization domain modifications, and designed a variety of RSV F protein mutants. Then, through gene synthesis, expression, and ELISA detection, the present invention tested the pre-fusion conformation (pre-F) ratio and stability under high and low temperature conditions of the RSV F protein mutants, and screened out several RSV F protein mutants with a relatively high pre-fusion conformation ratio and stable structure. Finally, candidate antigen representatives were selected for immunogenicity detection, which proved that they have good immunogenicity and can be used for the development of RSV vaccines, the preparation of RSV-specific antibodies, or RSV detection reagents.

The invention discloses a small cell piece treating agent for improving the culture quality of fresh water pearls and a treating method thereof, and relates to the technical field of aquaculture. The small cell piece treating agent is prepared by taking a phosphoric acid buffer salt solution as a basic solution, and functional components comprise a double-antibiotic solution and various growth factors; after fresh water pearl mussel cell slices are prepared, the cell slices are soaked in the treating agent for 3-5 minutes, and then nucleus insertion and pearl culture are carried out. By adopting the treatment agent and the treatment method, the problem of poor antibacterial effect of the monoclonal antibody can be solved when the pearl culture production of the freshwater pearl mussels is carried out, postoperative infection is effectively inhibited, and wound healing is promoted; the technical problem that the pearl bag forming speed is low is solved, pearl bag forming is accelerated, and pearl quality is improved. The product and the method disclosed by the invention are suitable for preoperative treatment of small cell pieces in a freshwater pearl mussel nucleus insertion operation, and have the characteristics of low cost, high stability and good effect.

The invention discloses a preparation method of a seawater shellfish single-cell suspension suitable for dynamic detection. The preparation method comprises the following steps: extracting gill filament tissues from living shellfish; washing gill filaments, and absorbing moisture; cutting the gill filament tissue into pieces; carrying out enzymolysis digestion on the gill filament tissue fragments by using an enzyme mixed solution consisting of collagenase II, hyaluronidase and deoxyribonuclease I; centrifuging and resuspending the cells; and filtering to obtain a single-cell suspension. Aiming at the condition that the shellfish tissue structure is relatively loose, cleaning liquid, enzyme liquid components, enzymolysis time, centrifugal speed and the like are optimized, and the high-quality single-cell suspension is obtained. According to the preparation method of the single-cell suspension, non-lethal sampling of seawater shellfish is conducted, the requirement for dynamic detection of shellfish traits in the industry can be met, enzymolysis is more sufficient, experiment time is shorter, operation steps are simple and convenient, obtained cells are more complete in morphology, the caking rate is extremely low, and the living cell rate is higher; and the requirements of construction of various single cell sequencing platform libraries can be fully met.

A cell culture medium that sustains the growth of cells obtained from a crustacean in vitro. Indeed, the disclosure describes a cell culture medium comprising a specific salt composition, in part responsible for the correct osmolality and pH, a specific amino acid and vitamin composition, energy sources such as glucose, and crustacean hemolymph. This disclosure further shows that the cell culture medium is capable of keeping cells obtained from shrimp and lobster viable for long periods of time even after passaging the cells into subcultures and inducing the cells to proliferate. The cultured cells are, for example, useful for performing physiological studies on certain genes (by genetic engineering) and for doing research and diagnosis on crustacean pathogens.

The invention discloses a prebiotic-loaded intestinal targeting exosome and a preparation method and application thereof.The preparation method comprises the steps that firstly, plasmids containing intestinal cell targeting peptide sequences are constructed through a gene editing technology, the plasmids are transfected and introduced into cells, the transfected cells are screened with antibiotics, and cell strains stably expressing intestinal cell targeting peptides are obtained; then culturing and adding prebiotics, and continuously culturing to enable the cells to take in the prebiotics and release the exosomes loaded with the prebiotics; and finally, collecting cell culture supernate, and centrifuging and resuspending for multiple times under a low-temperature condition to obtain the prebiotics-loaded intestinal targeting exosome. According to the preparation method, extraction of the exosome, loading of the prebiotics and an intestinal targeting modification process are optimized, so that the prepared intestinal targeting exosome loaded with the prebiotics can be used for remarkably improving the loading efficiency and intestinal targeting of the prebiotics and enhancing the stability of the exosome in gastrointestinal tracts; therefore, the delivery efficiency and the treatment effect of the prebiotics in the intestinal tract are improved.

The invention discloses a method suitable for extracting extracellular small vesicles in fresh water pearl shell tissue waste liquid, which comprises the following operation steps: (1) solid-liquid separation: slaughtering pearl shells, and filling slaughtering waste liquid into a waste liquid crude extraction device; (2) waste liquid crude extraction: performing multi-stage gradient filtration and gravity settling on the slaughtering waste liquid by using a waste liquid crude extraction device to obtain clarified waste liquid; (3) removing impurities: removing impurities for the first time and removing impurities for the second time; (4) concentration of the extracellular vesicles: concentrating the extracellular vesicles by using an ultra-speed centrifuge to obtain a high-concentration extracellular vesicle solution; and (5) preservation of extracellular vesicles: according to the final volume of the high-concentration extracellular vesicle solution obtained in the step (4), carrying out sample standardization through protein concentration, sub-packaging the sample, and carrying out long-term preservation in a low-temperature environment. The particle sizes of the extracted vesicles are concentrated, high-purity and complete extracellular small vesicles can be obtained, and the method is suitable for biomedical research and product development.

The present disclosure relates to the field of molecular virology, including nucleic acid molecules comprising a modified viral genome or self-replicating RNA, pharmaceutical compositions containing said nucleic acid molecules, and the use of such nucleic acid molecules and compositions for producing a desired product in a cell culture or in vivo. Also provided are methods for eliciting a pharmacodynamic effect in a subject in need thereof, as well as methods for preventing and / or treating a variety of health conditions.

The invention belongs to the field of gene engineering, and particularly relates to application of an RSAD1 gene / protein serving as a target spot in screening of drugs for inhibiting picornaviridae virus infection. The silent RSAD1 gene is accidentally found to be capable of remarkably inhibiting the replication of the Seneca virus and can be used as a target spot for researching and developing an antiviral drug for the Seneca virus. The invention provides an sgRNA sequence of a targeted RSAD1 gene, a CRISPR-Cas9 gene editing technology is combined, the RSAD1 gene is silenced, an RSAD1 gene knockout monoclonal cell line is obtained, the cell line can remarkably inhibit the replication of Seneca virus, and a target spot is provided for further screening drugs and reagents for resisting the replication of Seneca virus; meanwhile, it is found that overexpression of the RSAD1 protein in host cells can significantly promote replication of Seneca viruses, and the RSAD1 protein can be used as a production cell line of Seneca viruses or vaccines and used for production of Seneca viruses or vaccines.

The invention belongs to the field of biomedicine, and particularly discloses an influenza nanoparticle vaccine as well as a preparation method and application thereof. The fusion protein containing the influenza virus strain HA protein or the fragment thereof, the influenza virus protein epitope and the ferritin can be used as an antigen to induce an immune response effect aiming at influenza viruses, and a vaccine formed by matching the fusion protein with an MF59 adjuvant can be used for inducing to generate a relatively strong challenge protection effect and has a relatively good cross protection effect.

The invention relates to the technical field of bioengineering, in particular to an exosome separation and concentration method with marine animal source tissue as a raw material. Comprising the following steps: freeze-drying and crushing a marine animal source tissue sample; performing sample enzymolysis; and removing impurities and concentrating. According to the method disclosed by the invention, the exosome in the cells is fully dissociated through freeze-dried powder crushing and composite enzymolysis processes, and the filtering speed of the sample is accelerated by virtue of ultrasonic oscillation, so that the rapid and high-purity separation and concentration of the exosome in the marine animal source tissue are realized.

Disclosed in the present invention are an expression cassette containing overlapping open reading frames (ORFs), and a use. The overlapping ORFs are overlapping ORFs comprising a first ORF and a second ORF, and the expression cassette sequentially comprises from a 5' end to a 3' end: at least a first promoter used for driving transcription of a gene of the first ORF; a 5' part of the gene of the first ORF; an intron; and a 3'part of a gene of the second ORF, the intron comprising a second promoter only used for driving transcription of the gene of the second ORF. According to the present invention, two promoters are provided in a single expression cassette and are respectively used for driving expression of proteins of overlapping reading frames and adjusting the relative expression time and expression intensity of different proteins, thereby improving the production efficiency of rAAV vectors in insect cells, and also improving the quality of produced rAAV.

The invention belongs to the technical field of recombinant adeno-associated virus production, and discloses a method for improving the production efficiency of a recombinant adeno-associated virus and application. The method for improving the production efficiency of the recombinant adeno-associated virus comprises the following steps: adding a production promoting reagent into a cell culture solution; the production promoting reagent comprises at least one of a compound as shown in a formula (I) or a formula (II) and a derivative thereof. According to the method disclosed by the invention, the yield of rAAV produced by mammalian cells can be remarkably increased, so that the production cost is reduced, the large-scale production of rAAV is effectively promoted, and the method has important significance on rAAV-mediated gene therapy.

The invention relates to an RSV F protein mutant and application thereof, and belongs to the field of vaccines. On the basis of the amino acid sequence of wild RSV F protein, multiple aspects and multiple modes of modification are carried out, including disulfide bond mutation, proline mutation, cavity filling mutation, electrostatic mutation, p27 sequence modification and / or trimerization structural domain modification, and multiple RSV F protein mutants are designed. And then, through gene synthesis, expression and ELISA detection, the pre-conformation (pre-F) proportion of the RSV F protein mutants and the stability under high-temperature and low-temperature conditions are tested, and a plurality of RSV F protein mutants with relatively high pre-conformation proportion and stable structures are screened out. Finally, candidate antigen representatives are selected for immunogenicity detection, and it is proved that the candidate antigen representatives have good immunogenicity and can be used for developing RSV vaccines and preparing RSV specific antibodies or RSV detection reagents.

The invention discloses a culture method of in-vitro uniform monolayer cells of prawns, and belongs to the technical field of biology. The method comprises the following steps: (1) obtaining lymphatic tissues of a last-stage prawn at the early stage of shelling, completely stripping an outer membrane to obtain internal tissues, and cutting the internal tissues into small tissue blocks; (2) cleaning the small tissue blocks, inoculating the small tissue blocks to a culture plate coated with a gelatin solution, adding a complete culture solution into an inoculation system at intervals within 12 hours after inoculation for culture, replacing the complete culture solution 24-48 hours after inoculation, and inducing tissue cells to continuously migrate to form an in-vitro cell layer; (3) blowing and beating the cell culture to obtain a cell suspension, and centrifuging to obtain a cell monolayer; and (4) suspending the cell monolayer by adopting a complete culture solution, then inoculating the cell monolayer into a culture plate coated with a gelatin solution, and culturing to obtain the in-vitro uniform monolayer cells of the prawns. According to the method, the prawn monolayer cells which are long in in-vitro survival time, uniform and stable and are cultured in vitro can be obtained, and an important tool is provided for related virology research.

Provided is a detection technique excellent in cell drying resistance, cell retention, and detection of a chemical substance such as an odorant. More specifically, provided are: a laminate containing

The present invention provides a method for the production of a recombinant adeno-associated viral vector (rAAV), comprising contacting a host cell with a solution comprising at least one compound of formula (I), (IA), (IB), (II), (III), or (IV), or a salt thereof, or vitamin B, or any combination thereof. Also provided is a method for increasing production of rAAV by a host cell, comprising contacting the host cell with a solution comprising at least one compound of formula (I), (IA), (IB), (II), (III), or (IV), or a salt thereof, or vitamin B, or any combination thereof.

Provided are non-plant polypeptides or proteins comprising xylosylated N-linked glycans containing β1,2-xylose, wherein at least 25% of the individual protein species comprise xylosylated N-glycans, compositions comprising the non-plant proteins, cells engineered to produce the polypeptides or proteins, and methods for their production and use.

The present invention belongs to the field of insect cell culture technology and discloses a method for preparing an Sf9 cell feeder layer. The method mainly comprises two steps: step S1 is subculture of Sf9 cells, in which Sf9 cells are cultured in a suitable culture medium to a suitable cell confluence; step S2 is mitomycin C treatment of the Sf9 cells, in which a specific concentration of mitomycin C is used to treat the subcultured Sf9 cells at a specific time and temperature to inhibit their cell division ability, followed by washing to remove mitomycin C and replacing with fresh culture medium. The feeder layer cells obtained by the present invention can effectively inhibit their own proliferation while maintaining good cell activity and structural integrity. They can survive stably and provide necessary support for the co-culture system. They can be used for the cultivation of insect symbiotic bacteria and other difficult-to-culture insect cells. The method is simple to operate, has mild conditions, and good reproducibility. The prepared feeder layer has a stable effect, providing a powerful tool for related research fields.

To provide nucleic acid constructs, host insect cells, and methods for producing recombinant AAV capsids with high potency at high yield.SOLUTION: A nucleic acid construct comprises a nucleotide sequence encoding adeno-associated virus (AAV) VP1, VP2 and VP3 proteins, where the VP1 protein and the VP2 protein comprise two or more mutations at residues 157, 162, 164, 179, 188, 194, 196, 197, 200 and 201 in a specific sequence relative to wildtype VP1 and VP2 proteins.SELECTED DRAWING: None

The present disclosure provides a polypeptide fragment comprising a polypeptide of a modified hemagglutinin protein, wherein said modified hemagglutinin protein comprises a linker peptide replacing the polypeptide in the cytoplasmic domain of hemagglutinin protein, wherein the hemagglutinin protein is obtained from at least one seasonal strain of influenza virus, wherein said polypeptide of the modified hemagglutinin protein is having at least 80%, or at least 90%, or at least 95%, or at least 97% sequence identity to at least one sequence selected from the group consisting of SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6, SEQ ID NO: 8, SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14, SEQ ID NO: 16, SEQ ID NO: 18, SEQ ID NO: 20, SEQ ID NO: 22, SEQ ID NO: 24, SEQ ID NO: 26, SEQ ID NO: 28, and SEQ ID NO: 30.

The invention relates to the technical field of biology, and discloses a method for constructing a drosophila embryo permeabilization model, which comprises the following steps: collecting and cleaning drosophila embryos: collecting and washing the drosophila embryos; rinsing the drosophila melanogaster embryos, namely rinsing the drosophila melanogaster embryos by using a bleaching agent solution to remove chorion layers of the drosophila melanogaster embryos, and washing the embryos after rinsing; and performing permeation treatment on the drosophila melanogaster embryo: performing permeation treatment on the drosophila melanogaster embryo without the chorion layer in an embryo permeation solution EPS, and then cleaning in BPS to obtain the drosophila melanogaster embryo permeation model. According to the construction method of the drosophila melanogaster embryo permeabilization model, operation is easy, convenient and rapid, it can be guaranteed that embryo permeability is good, embryo vitality is maintained at the same time, the influence of small molecule drugs on the embryo development process is conveniently studied, and the method is suitable for popularization and application. And favorable conditions are provided for researching cell and molecular mechanisms of embryonic development, toxicological evaluation of small molecular substance screening and screening of related drugs in an embryo model.

Described herein are methods for making edible fat product using insect cells. The methods include isolating a population of insect cells, which may be precursor cells, seeding the population of insect cells onto a scaffold, and contacting the population of insect cells with a free fatty acid composition or a lipid composition. The cells seeded on the scaffold accumulate lipids, resulting in an edible fat tissue. Further disclosed are a composition of matter made from the method for making edible fat cells, a food product that includes the composition of matter, and an in vitro edible fat tissue comprising a population of insect precursor cells seeded on the scaffold.

Provided are an engineered mRNA and a method for improving protein expression using same. Also provided is the use of such engineered mRNA in gene therapy and / or genetic vaccination.

Provided is a method for culturing animal cells in which animal cells are cultured in the presence of an iron ionophore such as hinokitiol. Also provided is technology related to the method.