130results about "Invertebrate cells" patented technology

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 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 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 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.

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.

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.

The present disclosure provides a method for producing a methyl compound using glycine, serine, or an organic raw material.

The present invention belongs to the technical field of genetic engineering and cell engineering, and particularly relates to a rhabdovirus-negative Spodoptera frugiperda insect cell line, its screening, identification and use. According to the present invention, a rhabdovirus-negative Spodoptera frugiperda insect cell line WSK-Sf9 is obtained by screening and identification, and deposited under CCTCC NO:C 202246. The cell line is verified by various different highly sensitive assay methods such as nested PCR, transcriptome next generation sequencing, fluorescence-based quantitative PCR and probe-based quantitative PCR, and finally obtained by screening. The cells are tested for sterility, mycoplasma, exogenous viruses, tumorigenicity, etc. according to the requirements of the pharmacopoeia, and the results show that the cells meet the requirements in all tests. The cells can produce recombinant proteins based on the baculovirus expression system, and can be used for the production of recombinant protein vaccines.

The present invention belongs to the technical field of veterinary drug production, and provides a PCV2-SVA bivalent vaccine. The vaccine uses the PCV2 Cap protein with the nuclear localization signal removed and the SVA VP2 protein as antigens. The present invention efficiently expresses soluble PCV2 Cap protein and SVA VP2 protein through the baculovirus system. Due to the immune synergy, compared with commercial vaccines or inactivated vaccines, the PCV2-SVA bivalent vaccine prepared based on recombinant proteins can stimulate higher levels of humoral immunity and cellular immunity. The challenge protection test after immunization shows that the bivalent vaccine can effectively reduce the viremia and the virus content in organs. The bivalent genetic engineering vaccine against PCV2 and SVA developed based on the present invention has commercial development prospects.

The present disclosure relates to the field of molecular virology, including nucleic acid molecules comprising modified viral genomes or self-replicating RNAs, pharmaceutical compositions containing the same, and the use of such nucleic acid molecules and compositions for production of desired products in cell cultures or in a living body. Also provided are methods for eliciting a pharmacodynamic effect in a subject in need thereof, as well as methods for preventing and / or treating various health conditions.

The present invention relates to the technical field of genetic transformation of insect eggs. Specifically, the present invention refers to an efficient genetic editing system to obtain recombinant or genetically modified insect eggs, by incorporating genetic material directly into oocytes of female insects, which will then generate a large number of eggs with the incorporated or recombinant genetic material.

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 present invention provides a method of manufacturing a coral scaffold for use as a bone graft substitute. The method comprises growing coral in a growth medium having a carbonate hardness, dKH, of10 or more; removing at least a portion of the coral from the growth medium; devitalising coral removed from the growth medium and sizing the devitalised coral to form the coral scaffold.

This invention relates to a method for fabricating curved protein micropatterns, a field of materials and tissue engineering, involving the complex patterning of curved surfaces and the inner surfaces of tubular lumens, and the subsequent controlled cell arrangement. The method comprises: fabricating and surface treating a PDMS curved surface or tubular lumen substrate; tightly fitting a Parylene C porous membrane to the tubular lumen; incubating FN protein on the curved surface to create a complex protein pattern through infiltration; and seeding a cell sample at a predetermined density to achieve controlled cell arrangement and growth. This experiment can closely simulate the living environment of cells on complex curved surfaces and organs in the body, is simple to perform, and is inexpensive, providing new insights for studying cells on the surfaces of specialized tissues and organs.

Provided are a recombinant adeno-associated virus (rAAV) genome and a single-polarity rAAV (spAAV) vector formed by packaging same. Specifically, provided is an spAAV genome before packaging, two ends of which are both linear open terminuses, wherein one end does not have an inverted terminal repeat (ITR) or a part thereof, and the other end has a truncated ITR (ITRT), and the ITRT does not form a T-shaped hairpin palindromic structure. The spAAV genome is a single-polarity single-stranded DNA. Also provided are an spAAV vector, a method for delivering an exogenous nucleic acid to a cell by using the vector, and a use of the vector in the preparation of a product for gene expression, gene therapy, gene editing or gene regulation.

The present invention relates to the use of parasites and extracellular vesicles obtained from parasites for cancer treatment. The aim of the present invention is to use parasites and extracellular vesicles obtained from parasites in cancer treatment and to load active substances onto the exosomes by using the drug loading capacity of the exosomes and thus to carry the specific drug directly to the target cancer cells without causing any side effects on healthy cells and thereby to increase the bioavailability of the drug to achieve the desired effect in the tumor-specific target area. Within the scope of the present invention, in particular, the infantile leishmania parasite is used as a source of extracellular vesicles.

The present disclosure is related generally to systems and methods for high level expression of recombinant proteins from baculovirus in insect cells. In particular, the methods and systems described herein allow for high levels of baculovirus production in insect cells and / or high levels of protein production in insect cells using a chemically-defined, yeast lysate-free insect cell medium. The disclosure also relates to compositions and kits for culturing, transfecting, and / or producing recombinant protein in insect cells.

This invention belongs to the field of genetic engineering, specifically relating to the application of RSAD1 gene / protein as a target in screening drugs to inhibit infection by microribonucleoviridae viruses. This invention unexpectedly discovered that silencing the RSAD1 gene can significantly inhibit Seneca virus replication, and can be used as a target for the development of antiviral drugs against Seneca virus. This invention provides an sgRNA sequence targeting the RSAD1 gene, and combined with CRISPR-Cas9 gene editing technology, silences the RSAD1 gene, obtaining an RSAD1 gene knockout monoclonal cell line. This cell line can significantly inhibit Seneca virus replication, providing a target for further screening of drugs and reagents to resist Seneca virus replication. Simultaneously, this invention found that overexpression of the RSAD1 protein in host cells can significantly promote Seneca virus replication, and can be used as a production cell line for Seneca virus or vaccines.

The present invention provides a method of releasing viral vectors from cells producing those viral vectors by contacting the cells with a photosensitising agent which is then irradiated to disrupt the plasma membrane of the cells to release the viral vectors which may be collected and / or purified. The product of such methods as well as kits and apparatuses for performing the methods are also provided.

Provided herein are in vitro cultured fish compositions, methods and systems for culturing cells derived from fish sources.

The application provides a Marburg GP protein mutant, a DNA molecule, a recombinant vector and application thereof, and belongs to the technical field of Marburg virus vaccine preparation. The Marburg GP protein mutant is obtained by site mutation on the basis of a wild-type Marburg GP protein, and contains at least one mutation selected from R575P, V576P and L585P. The amino acid sequence of the wild-type Marburg GP protein is shown as SEQ ID NO:1. The Marburg GP mutant can provide a stable GP pre-fusion conformation, and can induce stronger binding activity, neutralizing activity and protective antibodies in mice, and is expected to be used for preparation of a Marburg virus vaccine.

To provide an adeno-associated virus vector (AAV) that can be very efficiently produced in insect cells.SOLUTION: The invention provides a nucleic acid construct comprising expression control sequences for expression in an insect cell of a nucleotide sequence comprising an open reading frame, the open reading frame sequence encodes: i) AAV capsid proteins VP1, VP2 and VP3; and ii) an ATG translation initiation codon for VP1. The nucleotide sequence also comprises, upstream of the open reading frame, an alternative start codon which is out of frame with the open reading frame.SELECTED DRAWING: None