67results about "Non-animal cells" patented technology

Cell culture apparatus comprising at least two adjacent cell cultivation channels separated by a permeable or semipermeable membrane, wherein at least one channel, for the majority of its length, has a cross sectional area of no more than 1 mm2, said channel being provided with entrance and exit means to permit the passage of media therethrough, allows co-culture of separate cell types, e.g. human and microbial cells, without mingling, allowing monitoring of cell cultures and chemical exchanges between the respective cell cultures.

The invention provides a fusobacterium nucleatum subsp.vincentii strain THCT14A3 for intestinal tract separation. The classification designation is Fusobacterium nucleatum subsp.vincentii, the preservation number is CCTCC NO:M 2019363, the preservation date is May 17th, 2019, and the preservation unit is China Center for Type Culture Collection. The 16SrRNA gene sequence of the strain is as shownin SEQID NO:1. The invention further provides a specific DNA sequence for identifying the fusobacterium nucleatum subsp.vincentii and a primer sequence corresponding to the specific DNA sequences. Thecolorectal cancer cell line and THCT14A3 co-culture result show that the THCT14A3 can significantly promote colorectal cancer cell proliferation. Therefore, the THCT14A3 provided by the invention canfacilitate micro-ecology research of intestinal tracts suffering from colorectal cancer.

The present invention it was determined that dendritic cells could be derived from various sources including peripheral blood monocytes in the presence of only GM-CSF without other cytokines if the monocytes were not activated. By preventing activation, such as by preventing binding of the cells to the surface of the culture vessel, the monocytes do not require the presence of additional cytokines, such as IL-4 or IL-13, to prevent differentiation into a non-dendritic cell lineage. The immature DCs generated and maintained in this manner were CD14− and expressed high levels of CD1a. Upon maturation by contact with an agent such as, for example, BCG and IFNγ, the cells were determined to express surface molecules typical of mature dendritic cells purified by prior methods and cultured in the presence of GM-CSF and IL-4. The mature dendritic cells produced from monocytes without activation and cultured in GM-CSF alone are suitable for use in dendritic cell-based immunotherapy methods, such as for use in the treatment of disease, including cancer.

The invention provides a fusobacterium nucleatum subsp.animalis strain THCT5A4 separated from human rectal cancer tumor tissue. A classification name is Fusobacterium nucleatum subsp.animalis THCT5A4,a preservation number of the fusobacterium nucleatum subsp.animalis strain THCT5A4 separated from the human rectal cancer tumor tissue is CCTCC NO: M 2019366, the preservation date is May 17th, 2019,the preservation organization is China General Microbiological Culture Collection Center, and a 16SrRNA gene sequence of the fusobacterium nucleatum subsp.animalis strain THCT5A4 separated from the human rectal cancer tumor tissue is shown in SEQ ID NO:1 as shown in the description. A separation method of the fusobacterium nucleatum subsp.animalis strain is provided. A result of co-culture of a colorectal cancer cell line and the THCT5A4 shows that the THCT5A4 can significantly promote proliferation of colorectal cancer cells, therefore, the THCT5A4 can provide diverse in-vitro or in-vivo experimental conditions for simulating an intestinal environment for studies of colorectal cancer, and a colorectal cancer disease model can be built for screening medicines for treating colorectal cancer.

The invention provides a fusobacterium nucleatum polymorphic subspecies isolate THCT15E1, a classification name is Fusobacterium nuucleates subsp.nucleatum THCT15E1, a preservation number of the strain is CCTCC NO: M 2019362, a preservation date is May 17, 2019, a preservation unit is China Center for Type Culture Collection, and the 16SrRNA gene sequence of the strain is shown as SEQ ID NO: 1. The result of co-culture of the colorectal cancer cell line and THCT15E1 shows that THCT15E1 can significantly promote colorectal cancer cell proliferation; therefore, the THCT15E1 can provide diversified experimental conditions for in-vitro or in-vivo intestinal environment simulation for colorectal cancer research, and can also construct a colorectal cancer disease model to screen drugs for treating colorectal cancer.

The invention discloses a soybean bHLH transcription factor gene GmFER and encoded protein and application thereof. The nucleotide sequence of the gene GmFER is as shown in SEQ ID No.1, and the amino acid sequence of the protein encoded by the gene is as shown in SEQ ID No. 2. A semi-quantitative detecting result shows that the gene GmFER is mainly expressed in soybean roots; the expression quantity of the gene GmFER is increased evidently after Cd stress, that is to say, the gene GmFER is induced by Cd; by overexpressing the gene GmFER in yeast and plants, the Cd tolerance of the yeast and plants can be increased, and the gene GmFER is high in application value.

A microorganism co-culture system, comprising:(1) a substrate, comprising a saccharide(2) at least one of a first strain and a second strain, wherein the first strain is able to fix a carbon oxide the second strain is able to fermentatively metabolize an amino acid, and wherein the first strain produces a first metabolite in the fermentation, and the second strain produces a second metabolite in the fermentation; and(3) a third strain, being able to metabolize the saccharide, the first metabolite and the second metabolite in the fermentation to produce butyric acid and / or butanol,wherein, when the second strain is present in the co-culture system, the substrate further comprises an amino acid.

The purpose of the present invention is to provide a composition for promoting the proliferation of a fibroblast and / or a composition for promoting the expression of a hyaluronic acid synthase gene. The present invention provides a composition containing Lactobacillus plantarum L-137 and used for promoting the proliferation of the fibroblast and / or the composition for promoting the expression of the hyaluronic acid synthase gene.

Provided are compositions for long-term maintenance of functional hepatocytes in culture, a method for improved maintenance of functional hepatocytes in vitro, and functional hepatocytes cultures according to the methods. The culture compositions include at least: one activator of adenylate cyclase, one TGFbeta inhibitor, one Notch inhibitor, one Wnt inhibitor, and / or one BMP inhibitor. The combinations of compounds are added to any hepatocyte cell culture medium in an effective amount to maintain functional hepatocyte function in vitro, long term. The hepatocytes can be used for in vitro drug research and to model liver disease.

It is an object of the present invention to provide a method for producing pluripotent cells that are free of the risk of cellular canceration and that can be applied to regenerative medicine with a high degree of safety. The present invention provides a method for producing pluripotent cells from somatic cells comprising a step of bringing bacteria having fermentation ability or a component or secretory product thereof into contact with somatic cells.

The present invention it was determined that dendritic cells could be derived from various sources including peripheral blood monocytes in the presence of only GM-CSF without other cytokines if the monocytes were not activated. By preventing activation, such as by preventing binding of the cells to the surface of the culture vessel, the monocytes do not require the presence of additional cytokines, such as IL-4 or IL-13, to prevent differentiation into a non-dendritic cell lineage. The immature DCs generated and maintained in this manner were CD14− and expressed high levels of CD1a. Upon maturation by contact with an agent such as, for example, BCG and IFNγ, the cells were determined to express surface molecules typical of mature dendritic cells purified by prior methods and cultured in the presence of GM-CSF and IL-4. The mature dendritic cells produced from monocytes without activation and cultured in GM-CSF alone are suitable for use in dendritic cell-based immunotherapy methods, such as for use in the treatment of disease, including cancer.

The invention discloses a method for greatly increasing the PCV2 cultivation virus titer through a streptococcus culture preparation, and belongs to the technical field of cell engineering and virus cultivation. The streptococcus culture preparation is prepared through high-temperature inactivation of streptococcus-containing supernate which is obtained after co-culture of streptococcus and anchorage-dependent cells in a maintenance culture medium. The streptococcus is streptococcus agalactiae, the cells are PK-15 cells, and the content of the streptococcus in the streptococcus-containing supernate is 100-900 million / mL. The streptococcus culture preparation is added into the maintenance culture medium according to the concentration of 0.1-1% to prepare a special culture medium used for stimulating PCV2 to generate viruses, the cultivation virus titer of the culture medium can be greatly increased by adopting the special culture medium as the maintenance culture medium for culturing PCV2, and the virus titer can reach 10<-7> or above; and a production process is greatly simplified, and the production cost is greatly lowered.

A method of co-culturing helicobacter pylori and gastric mucosa epithelial cells is disclosed. The method includes 1) preparing a helicobacter pylori medium; 2) adding the medium into a sterile cell culture flask and preparing a slant surface; 3) culturing gastric mucosa epithelial cells in the culture flask in which the slant surface is formed and allowing the cells to be adhered to the wall of the flask; 4) centrifuging a helicobacter pylori 48-h culture liquid, and then resuspending a centrifuging product with a cell culture liquid; and 5) adding a resuspended bacterial liquid into the culture flask in which the slant surface and the adherent cells are formed, culturing, and counting the living bacteria and observing cell morphological changes every day. A beneficial effect of the method is that the novel method of co-culturing the helicobacter pylori and the gastric mucosa epithelial cells is provided. In the novel system, the helicobacter pylori can grow normally and continuously stimulate the adherent cells to generate lesions, thus simulating a human gastric mucosa infection situation well. The problem that helicobacter pylori declines or dies and cells are not effectively stimulated because culture liquids are not suitable for growth of the helicobacter pylori in traditional methods in which helicobacter pylori are directly added is overcome.

The present invention relates to a method for the preparation of an immunogenic composition for the treatment and / or prophylaxis of mycoplasma infections in a subject comprising the cultivation of mycoplasma bacteria in a serum-reduced or swine serum-free, eukaryotic cell system; obtaining an antigen of the mycoplasma bacteria; and addition of a pharmaceutically acceptable carrier. Further, the present invention relates to the immunogenic composition obtainable by said method and a method for immunizing a subject comprising the administration of said immunogenic composition to a subject.

Numerous plant microbes, including the vascular-limited Candidatus spp.—causal agents of citrus greening and potato zebra chip diseases—are non-culturable. The present disclosure relates, according to some embodiments, to compositions, methods and systems for culturing such organisms. For example, the present disclosure relates to methods for culturing, propagating, and characterizing fastidious vascular-colonizing microbes using a hairy root system (e.g., in vitro, in planta). The present disclosure relates, in some embodiments, to methods for cultivating a fastidious plant microbe including: contacting a plant (e.g., a tomato plant, a potato plant, a citrus plant) colonized by a fastidious plant microbe (e.g., Xylella fastidiosa, Candidatus Liberibacter spp.) with a suspension of R. rhizogenes under conditions that permit induction of hairy roots colonized with the fastidious plant microbe, and propagating the colonized microbial hairy roots.

The invention relates to a preparation method and an application of a cell membrane coated nano topological structure array. The preparation method comprises the following steps: stimulating macrophages to form stimulated macrophages, and extracting stimulated macrophage membranes; meanwhile, processing the substrate to form a substrate with nanowires, and processing the substrate with the nanowires to form a nanowire substrate with positive charges; and combining the stimulated macrophage membrane with the nanowire substrate with the positive charges to obtain the macrophage membrane modifiednano topological structure array. The cell membrane coated nano topological structure array is simple in preparation and operation, and can be applied to capture of bacteria.

An inhibiting or alleviating agent for inflammation in the brain comprising an extract from inflamed tissue inoculated with vaccinia virus as the active ingredient. A determination or evaluation method of an extract from inflamed tissue inoculated with vaccinia virus or an agent comprising the extract, characterized in that the inhibition of the expression of pro-inflammatory cytokines and / or NF-kB pathway related proteins induced by the promotion of expression of BDNF in cultivated glial cells is used as an indicator. A use of an extract from inflamed tissue inoculated with vaccinia virus in the production of the inhibiting or alleviating agent for inflammation in the brain.

Described is a method to transfer chromosomal DNA between two microbial species without genetic engineering or vectors. The strains resulting from this method are chimeric microbial hybrids that can express a combination of genotypes from both parents.

Hepatitis C virus replication at extrahepatic sites has been suggested; however, complete viral replication has only been confirmed in hepatocytes. Here we show that human adipogenic DLK-1+ stem cells (hADSC) freshly isolated from HCV-infected individuals contained viral transcripts, replication intermediates and viral antigens in vivo, and viral transcripts increased in supernatants upon prolonged ex vivo culture. Furthermore, naive hADSC isolated from HCV (−) individuals support complete replication of clinical isolates in vitro, and the infection is donor-nonspecific for cells and cross-genotypic for viruses. Viral infection / replication is mediated through CD81, LDL-R, SR-B1, EGFR, Apolipoprotein E, occludin, claudin-1, NPC1L1 and diacylglycerol acetyltransferase-1, and can be inhibited by anti-viral drugs. In addition, the physical properties of hADSC-propagated viral particles resemble clinical isolates more than JFH1 / HCVcc, and viruses propagated by in vitro infected hADSC are infectious to primary human hepatocytes. Therefore, hADSC are an in vivo HCV reservoir and represent a novel venue of clinical virus-host interaction. hADSC can also be exploited as a physiologically relevant primary cell culture system to propagate clinical isolates.

It is an object of the present invention to provide a method for producing pluripotent cells that are free of the risk of cellular canceration and that can be applied to regenerative medicine with a high degree of safety. The present invention provides a method for producing pluripotent cells from somatic cells comprising a step of bringing bacteria having fermentation ability or a component or secretory product thereof into contact with somatic cells.

The invention belongs to the technical field of biological medicine, and particularly relates to a culture medium and method for constructing a liver cancer or bile duct cancer explant and application. According to the culture medium and the method for constructing the liver cancer or cholangiocarcinoma explant, tumor tissues cut by a liver cancer patient through an operation are utilized, an optimized explant culture medium is utilized for multiplication culture, and a liver cancer explant model with high activity and a complete structure can be constructed through construction of an explant culture method; the method can be used for screening and guiding drugs.

Hepatitis C virus replication at extrahepatic sites has been suggested; however, complete viral replication has only been confirmed in hepatocytes. Here we show that human adipogenic DLK-1+stem cells (hADSC) freshly isolated from HCV-infected individuals contained viral transcripts, replication intermediates and viral antigens in vivo, and viral transcripts increased in supernatants upon prolonged ex vivo culture. Furthermore, naive hADSC isolated from HCV(-)individuals support complete replication of clinical isolates in vitro, and the infection is donor-nonspecific for cells and cross-genotypic for viruses. Viral infection / replication is mediated through CD81, LDL-R, SR-B1, EGFR, Apolipoprotein E, occludin, claudin-1, NPC1L1 and diacylglycerol acetyltransferase-1, and can be inhibited by anti-viral drugs. In addition, the physical properties of hADSC-propagated viral particles resemble clinical isolates more than JFH1 / HCVcc, and viruses propagated by in vitro infected hADSC are infectious to primary human hepatocytes. Therefore, hADSC are an in vivo HCV reservoir and represent a novel venue of clinical virus-host interaction. hADSC can also be exploited as a physiologically relevant primary cell culture system to propagate clinical isolates.

Described is a method to transfer chromosomal DNA between two microbial species without genetic engineering or vectors. The strains resulting from this method are chimeric microbial hybrids that can express a combination of genotypes from both parents.

The invention discloses an NKT cell in-vitro amplification method based on a combined stimulant, and belongs to the technical field of biology. The method comprises the following steps: firstly separating mononuclear cells in autologous peripheral blood, and then performing induced activation and amplification on the NKT cells in a serum-free culture system by adding a group A streptococcus preparation, a zoledronic acid preparation and a cytokine IL-2 as a combined stimulant. The method is easy to operate, efficient amplification of the NKT cells can be achieved only through three key reagents, the obtained NKT cells have the advantages of being large in number, high in purity, high in cytotoxicity and the like, and the requirements of clinical treatment can be met. Compared with a traditional method, the culture cost is remarkably reduced, the operation process is simplified, large-scale production, popularization and application are easy, and an efficient and economical technical scheme is provided for NKT cellular immunotherapy.

Provided is a use of dipsaci radix-derived extracellular vesicle-like nanoparticles (DREVNs) in preparation of a drug for preventing or treating orthopedic diseases. In this application, EVs are creatively extracted from dipsaci radix and purified, and the physiological efficacy of the EVs is studied. It has been found that the EVs can be fully internalized by bone marrow mesenchymal stem cells (BMSCs), can promote the osteogenic differentiation of BMSCs by activating a BMP2 / Smads signaling pathway, promote the calcified nodule formation in BMSCs, and promote the expression of osteogenic differentiation-associated genes ALP, OCN, RUNX2, and COL1, and have bone targetability in vivo. The EVs can be intragastrically administered to alleviate the osteoporosis (OP) in postmenopausal mice. Therefore, the EVs have the potential of being used to prepare drugs for preventing or treating orthopedic diseases, and provide a new strategy for the prevention or treatment of orthopedic diseases.

An object of the present invention is to obtain a thicker animal cell composition by a simple and less expensive method. Namely, an object of the present invention is to provide a method for culturing a thicker animal cell composition by eliminating the hypoxia associated with animal cell compositions, a method for producing an animal cell composition containing unicellular algae, and an animal cell composition.The present invention provides a method for culturing an animal cell composition in a culture medium in the presence of unicellular algae and under exposure to light. According to the method of the present invention, oxygen can be continuously supplied in the culture medium, cell damage is alleviated, and a thicker cell composition can be obtained in the absence of a capillary network.

A culture container for culturing epithelial cells that comprises an upper container, a lid member air tightly fitting into the opening of the upper container, and a lower container housing the upper container and a cell culture medium therein, wherein; at least a part of an area of the upper container, said area being in contact with the cell culture medium, is formed of a membrane that is permeable to at least some components of the cell culture medium and impermeable to cells; and the material of the lid member has an oxygen permeation coefficient of not more than 1.0x10<-6> cm<3> cm / (cm<2> s.Pa).

An inhibiting or alleviating agent for inflammation in the brain comprising an extract from inflamed tissue inoculated with vaccinia virus as the active ingredient. In another aspect, the invention relates to a determination or evaluation method of an extract from inflamed tissue inoculated with vaccinia virus or an agent comprising the extract, characterized in that the inhibition of the expression of pro-inflammatory cytokines and / or NF-κB pathway related proteins induced by the promotion of expression of BDNF in cultivated glial cells is used as an indicator. In still another aspect, the invention also relates to a use of an extract from inflamed tissue inoculated with vaccinia virus in the production of the inhibiting or alleviating agent for inflammation in the brain.

The invention discloses a composition containing ginseng exosome and super-elastic protein and application of the composition, and belongs to the technical field of biology. The composition containing the ginseng exosome and the super-elastic protein is prepared by compounding the ginseng exosome and the super-elastic protein according to the weight ratio of 10: (100-150), the ginseng exosome is prepared by the following method: extracting the ginseng exosome from ginseng root tissues by using a differential centrifugation method to obtain a ginseng exosome crude extract; adding sucrose solutions with different concentrations into the ginseng exosome crude extract, and performing ultra-high-speed centrifugal purification to obtain purified ginseng exosome; the amino acid sequence of the super-elastic protein is as shown in SEQ ID NO. 3. According to the invention, the ginseng exosome and the super-elastic protein are combined for use for the first time, and both the ginseng exosome and the super-elastic protein have the effect of promoting cell proliferation, so that the use safety is high; the ginseng exosome and the super-elastic protein can synergistically improve the skin tightening and anti-wrinkle effects and the skin injury repairing effect.

Disclosed herein is a Chlamydia-activated B cell (CAB) platform. Also disclosed is a method of enhancing a population of B cells, comprising exposing said B cells to Chlamydia spp. under conditions suitable to enhance the population of B cells, such that expansion and differentiation of said B cells takes place, and said B cells are exposed or crosslinked to an antigen. Also disclosed are methods of producing said CABs, and treating a subject in need thereof with said CABs.