102 results about "Coenocyte" patented technology

The present invention provides a system for enhancing clearance or destruction of undesirable cells or noncellular molecular entities by tagging such cells or noncellular molecular entities with a marker that targets the cells or noncellular molecular entities for phagocytosis (phagocytic marker). The target cells can be, for example, endothelial cells, tumor cells, leukocytes, or virus-infected cells. In certain embodiments of the invention the tagging is accomplished by administering a composition comprising an antibody or ligand linked to the phagcytotic marker, wherein the antibody or ligand binds to a cell type specific marker present on or in the cell surface of a target cell. In preferred embodiments of the invention, the phagocytic marker comprises phosphatidylserine or a group derived from phosphatidylserine, thrombospondin-1, annexin I, or a derivative of any of these.

The present invention provides methods and compositions for isolating and detecting rare cells from a biological sample containing other types of cells. In particular, the present invention includes a debulking step that uses a microfabricated filters for filtering fluid samples and the enriched rare cells can be used in a downstream process such as identifies, characterizes or even grown in culture or used in other ways. The invention also include a method of determining the aggressiveness of the tumor or of the number or proportion of cancer cells in the enriched sample by detecting the presence or amount of telomerase activity or telomerase nucleic acid or telomerase expression after enrichment of rare cells. This invention further provides an efficient and rapid method to specifically remove red blood cells as well as white blood cells from a biological sample containing at least one of each of red blood cells and white blood cells, resulting in the enrichment of rare target cells including circulating tumor cells (CTC), stromal cells, mesenchymal cells, endothelial cells, fetal cells, stem cells, non-hematopoietic cells etc from a blood sample. The method is based upon combination of immuno-microparticles (antibody coated microparticles) and density-based separation. The final enriched target cells can be subjected to a variety of analysis and manipulations, such as flowcytometry, PCR, immunofluorescence, immunocytochemistry, image analysis, enzymatic assays, gene expression profiling analysis, efficacy tests of therapeutics, culturing of enriched rare cells, and therapeutic use of enriched rare cells. In addition, depleted plasma protein and white blood cells can be optionally recovered, and subjected to other analysis such as inflammation studies, gene expression profiling, etc.

The present invention relates to adenovirus E4ORF 1 gene and to endothelial cells engineered to express the E40RF 1 gene. The present invention also relates to uses of the E40RF 1 gene, and cells expressing the E40RF1 gene, and to compositions comprising the E4ORF 1 gene, or comprising cells expressing the E4ORF 1 gene.

Extracts from plant material, or semi-purified / purified molecules or compounds prepared from the extracts that demonstrate the ability to modulate one or more cellular activities are provided. The extracts are capable of slowing down, inhibiting or preventing cell migration, for example, the migration of endothelial cells or neoplastic cells and thus, the use of the extracts to slow down, inhibit or prevent abnormal cell migration in an animal is also provided. Methods of selecting and preparing the plant extracts and methods of screening the extracts to determine their ability to modulate one or more cellular activity are described. The purification or semi-purification of one or more molecules from the described extracts is also contemplated as well as the use of these molecules, alone or in combination with an extract, to slow down, inhibit or prevent abnormal cell migration in an animal.

The invention aims at providing a novel full-thickness biological cornea used for transplantation. The full-thickness biological cornea takes an animal cornea acellular matrix as a carrier, and the acellular matrix comprises animal cornea matrix cells, epithelial cells and endothelial cells which are cultured and augmented in vitro. The cornea is characterized in that the xenogenic corneal acellular matrix prepared by a biochemical method can be used as a good carrier for in vitro constructing the biological cornea in the aspects of shape, structure and biological compatibility, the matrix cells, epithelial cells and endothelial cells of the cornea are planted respectively, and dynamically cultured in the simulated in vivo environment of a biological reactor, so that the full-thickness biological cornea with nearly normal tissue structure and characteristics can be constructed in vitro, and the biological cornea can be used to simulate the physiological cornea for fundamental research on physiology, pathology and pharmacology; moreover, the biological cornea can also be directly used as the donator for corneal transplantation.

An in vitro biomechanical model used to applied hemodynamic (i.e., blood flow) patterns modeled after the human circulation to human / animal cells in culture. This model replicates hemodynamic flow patterns that are measured directly from the human circulation using non-invasive magnetic resonance imaging and translated to the motor that controls the rotation of the cone. The cone is submerged in fluid (i.e., cell culture media) and brought into close proximity to the surface of the cells that are grown on the plate surface. The rotation of the cone transduces momentum on the fluid and creates time-varying shear stresses on the plate or cellular surface. This model most closely mimics the physiological hemodynamic forces imparted on endothelial cells (cell lining blood vessels) in vivo and overcomes previous flow devices limited in applying more simplified nonphysiological flow patterns. Another aspect of this invention is directed to incorporate a transwell co-cultured dish. This permits two to three or more different cell types to be physically separated within the culture dish environment, while the inner cellular surface is exposed to the simulated hemodynamic flow patterns. Other significant modifications include custom in-flow and out-flow tubing to supply media, drugs, etc. separately and independently to both the inner and outer chambers of the coculture model. External components are used to control for physiological temperature and gas concentration. The physical separation of adherent cells by the artificial transwell membrane and the bottom of the Petri dish permits each cell layer, or surface to be separately isolated for an array of biological analyses (i.e., protein, gene, etc.).

The invention relates to a high-performance angiogenesis inhibitor and a production method, which belongs to the field of the biological engineering pharmaceutical technology or protein polypeptide drugs. The invention designs a high-performance angiogenesis inhibitor RGD-ED with integrin compatibility, the inhibitor contains angiogenesis inhibition polypeptide isoleucine-valine-arginine-arginine-alanine-aspartate-arginine-alanine-alanine-valine-proline, and one or two ends of the inhibitor are respectively connected with polypeptides containing arginine-glycin-aspartate sequence. The RGD-ED of the invention can be synthesized. By the method of genetic engineering, the invention also expresses one of RGD-Eds in escherichia coli or other eukaryotic cells, and the RGD-ED is obtained by carrying out the separation, dissolution and renaturation of inclusion body protein and separation and purification by ion exchange chromatography. All the polypeptide sequences of the invention are modified by Polyethylene Glycol (PEG), heparin, dextran, polyvinylpyrrolidone (PVP), polyglycol-poly (amino acid) copolymer, palmitic acid, colominic acid and liposome, which includes liposome (REV), drying liposomal (DRV) and multivesicular liposome (Mvl). In vivo and in vitro experiments, the synthetic polypeptide sequence, product of genetic engineering and modified product of the invention can notably increase the effects of inhibiting the growth of endothelial cells, inhibiting angiogenesis and resisting tumor of the present angiogenesis inhibitors, and moreover, the high-performance angiogenesis inhibitor can be used as a drug curing solid tumors and rheumatoid arthritis.

The present invention relates to adenovirus E4ORF 1 gene and to endothelial cells engineered to express the E40RF 1 gene. The present invention also relates to uses of the E40RF 1 gene, and cells expressing the E40RF1 gene, and to compositions comprising the E4ORF 1 gene, or comprising cells expressing the E4ORF 1 gene.

The invention relates to an HUVEC (human umbilical vein endothelial cell) separation, culture and subculture method belonging to the technical field of biology. The method comprises the following steps: (1) separation of HUVEC: taking a fresh umbilical cord, cleaning with a sterile PBS (phosphate buffer solution) containing penicillin-streptomycin, adding collagenase to perform water bath digestion at 37 DEG C, centrifuging the digestion solution, then adding an M199 culture medium, transferring into a gelatin-coated culture bottle, and culturing in a CO2 incubator at 37 DEG C; (2) culture of HUVEC: after the HUVEC is cultured at 37 DEG C for 24 hours, pouring out the culture medium, cleaning with the PBS, adding a fresh M199 culture medium, and afterwards, changing the culture medium once every two days, wherein the HUVEC can be subcultured generally after being cultured for 5-7 days; and (3) subculture of HUVEC: pouring out the culture medium, cleaning with the PBS, adding the digestion solution to digest the cell, adding a DMEM (dulbecco's modified eagle medium) containing serum to terminate the reaction once the cell is rounded, centrifuging the digested cell, and adding a fresh culture medium, wherein the HUVEC subcultured for 2-3 generations is used for experiments. The separated HUVEC is economical and practical, is simple and easy to use, and is beneficial to obtaining required in-vitro experimental model cells.

The invention discloses a construction method and application of a retinal neovascularization disease model, and relates to the field of biotechnology. The construction method knocks out the Tmem30a gene sequence in the genome of the target animal vascular endothelial cells, so that the target animal exhibits the typical retinal neovascularization disease characteristics. The target animal can beused as a model of retinal neovascularization disease, which can help to clarify the pathogenesis and mechanism of retinal neovascularization disease, and provide a new target for the treatment or prevention of retinal neovascularization disease.

The invention discloses a decellularized method for cornea. The decellularized method includes the following steps that (A), the picked animal cornea is placed in water, vibration treatment is carried out, and an epithelial layer falls off accordingly; (B), the cornea is placed in decellularized reagents for vibration treatment; (C), the cornea is placed in water for vibration treatment; (D), the step B and the step C are repeated alternately so that stroma cells and endothelial cells can fall off, wherein the decellularized reagents contain NaCl and EDTA. According to the decellularized method, the epithelial layer of the cornea and immunogen cellular constituents can be removed rapidly and tenderly, and therefore undamaged decellularized cornea ground substances can be obtained and have good biocompatibility and anti-degradation capacity.

Methods for predicting patient tolerability to therapeutic agents, such as cytokines, lymphokines and immunotoxins, are disclosed. The methods utilize an in vitro model of vascular leak syndrome (VLS) to assess the effect of the agent in question on the permeability of large proteins across confluent monolayers of endothelial cells (EC).

An isolated composition of matter is provided comprising a heterogeneous population of cells seeded on a surface of a scaffold, wherein the heterogeneous population of cells comprises at least one pancreatic islet, endothelial cells and fibroblast cells. Methods of generating same and uses thereof are also provided.

The present invention provides a system for enhancing clearance or destruction of undesirable cells or noncellular molecular entities by tagging such cells or noncellular molecular entities with a marker that targets the cells or noncellular molecular entities for phagocytosis (phagocytic marker). The target cells can be, for example, endothelial cells, tumor cells, leukocytes, or virus-infected cells. In certain embodiments of the invention the tagging is accomplished by administering a composition comprising an antibody or ligand linked to the phagcytotic marker, wherein the antibody or ligand binds to a cell type specific marker present on or in the cell surface of a target cell. In preferred embodiments of the invention, the phagocytic marker comprises phosphatidylserine or a group derived from phosphatidylserine, thrombospondin-1, annexin I, or a derivative of any of these.

The present invention provides a method for subculturing primate embryonic stem cells, and a method for inducing differentiation of the same cell into a vascular endothelial cell and a blood cell. The present invention provides a method comprising culturing primate embryonic stem cells in a medium containing a protein component without using feeder cells and cytokines in a container coated with an extracellular matrix, detaching colonies of the resulting embryonic stem cells in the presence of a cytodetachment agent, and plating the colonies in the similar medium, and a method comprising culturing primate embryonic stem cells in a serum-containing or not containing medium in the presence of cytokine, adhesion-culturing the resulting embryoid body or embroyid body-analogous cellular aggregate in the presence of a cytokine to obtain specific precursor cells, and separating non-adherent cells and adherent cells from the specific precursor cells to obtain blood cells and vascular endothelial precursor cells.

The present invention proposes formative agent of protein complex, in which a polyphenol is useful component, and the agent is useful as gene complex, cell adhesion inhibitor or immune tolerogen. The polyphenol of forming the agent is selected from catechin group consisting of epigallocatechin-gallate, tannic acids, or proanto-dianisidine, a protein of the protein complex is selected from proteins consisting of animal proteins composed of polypeptide chain of peptide-combined amino acids, vegetative proteins, nucleus proteins, glycogen proteins, lipo-proteins and metal proteins, the gene complex comprises by compositing genes by polyphenol catechins in order to introduce genes to cells of animals or human bodies, a cell composed of the cell adhesion inhibitor is selected from cells consisting of an animal cell including a stem cell, skin cell, mucosa cell, hepatocyte, islet cell, neural cell, cartilage cell, endothelial cell, epidermal cell, osteocyte or muscle cell isolated from human or animal organism, or sperm, ovum or fertilized egg of domestic animals or fishes and a tissue or an organ for transplantation of the immune tolerogen is selected from the tissue or the organ consisting of skin, blood vessel, cornea, kidney, heart, liver, umbilical cord, bowels, nerve, lung, placenta or pancreas.

The invention provides reduction-responsive hyperbranched poly-beta-amino ester with efficient gene delivery capability and a preparation method and application thereof. The polymer is polymerized byan A2 + B3 + C2 Michael addition method so that the polymer has a hyperbranched structure. Compared with a linear structure, the branched structure can enhance the interaction between the polymer andnucleic acid molecules, significantly improve the gene condensation ability of the polymer, and increase cellular uptake by enhancing the interaction with cell membranes. The main chain of the polymerhas a reduction-responsive group (disulfide bond), and under the action of GSH in damaged vascular endothelial cells, poly-beta-amino ester can be quickly degraded in cytoplasm to release an entrapped gene drug (ICAM-1 siRNA) so that efficient transfection of genes is realized, and meanwhile, the toxicity of the material is reduced. Due to the properties of the poly-beta-amino ester, the poly-beta-amino ester has a huge development prospect in biomedical materials, especially in the field of gene delivery.

The invention relates to platycodon grandiflorum polysaccharide extracted from rhizome of platycodon grandiflorum, a degradation product of the polysaccharide, a method for extracting the platycodon grandiflorum polysaccharide from the rhizome of the platycodon grandiflorum, a method for preparing the degradation product of the platycodon grandiflorum polysaccharide, and application of the platycodon grandiflorum polysaccharide and the degradation product thereof to preparing a medicine for treating anti-tumour cell angiogenesis diseases. The structure of the platycodon grandiflorum polysaccharide disclosed by the invention is (1->4)-alpha-D- pyranose homogalacturonan, has the polymerization degree of 15-100, the corresponding molecular weight of 2.0-18.0 kD, and the specific rotation of [alpha]D19+125 DEG (c0.05, H2O), and is represented by the following structural formula described in the specification. The molecular weight range of a degradation product obtained by partial acid hydrolysis of the platycodon grandiflorum pectin polysaccharide is 1.4-3.0 kDa. Vivo experiments prove that the platycodon grandiflorum polysaccharide can obviously inhibit lumen generation function of a human microvascular endothelial cell (HMEC-1) and is hopefully used as a novel anti-tumour medicine for inhibiting the growth of a solid tumour by inhibiting angiogenesis.

The invention relates to a medicine for in vitro hemostasis and a preparation method of the medicine. The in vitro injury bleeding conditions frequently occur in life; some in vitro injury bleeding conditions are caused by chance; some in vitro injury bleeding conditions are generated by combination with internal causes of the organism; the bleeding is caused by vascular permeability increase of postcapillary venule, capillary vessels and precapillary arteriole; blood leaks out of a lumen through an expanded endothelial cell clearance and a damaged vascular basement membrane, for example, finger injury bleeding, toe injury bleeding, skin scratch bleeding and the like. The medicine is prepared by combining the following raw materials: cirsium japonicum, cuttle bone, herba cirsii, cutch, sanguisorba officinalis, dragon blood, ramie root, loofah sponge, sophora japonica, rheum officinale, puffball, hairyvein agrimony, shrubalthea bark, rhizoma bletillae, alumen, crinis trachycarpi, coptis chinensis, folium artemisiae argyi, indigo naturalis, panax notoginseng, carbonized hair, pericarpium granati, cattail pollen, dark plum, ophicalcite, mongolian snake gourd roots, selaginella tamariscina, halloysitum rubrum, schizonepeta and cacumen biotae. The raw materials are combined to generate the synergistic effect in efficacy, so that in vitro injury bleeding can be effectively treated.

Provided is a means for reconstructing tissues and organs having mature functions. The method for producing tissues and organs is characterized by co-culturing organ cells with vascular endothelial cells and undifferentiated mesenchymal cells, producing an organ bud, and transplanting the same into a non-human animal, after which the tissue or organ derived from the transplanted organ bud is extracted from the non-human animal.

The invention provides a euphausia superba-originated antihypertensive tripeptide and application thereof and belongs to the technical field of antihypertensive oligopeptides. The euphausia superba-originated antihypertensive tripeptide has an amino acid sequence of Phe-Ala-Ser with an ESI-MS (electrospray ionization mass spectrometry) measured molecular weight of 323.35 Da. The preparation methodof the euphausia superba-originated antihypertensive tripeptide comprises homogenizing euphausia superba, adding in distilled water and proteases for enzymolysis in an environment of 45-55 DEG C for5.5-6.5 h to obtain an enzymolyzed euphausia superba solution; performing ultrafiltration, anion exchange chromatography, gel column chromatography and reversed-phase high-performance liquid chromatography to prepare the euphausia superba-originated antihypertensive tripeptide. The euphausia superba-originated antihypertensive tripeptide achieves significant inhibiting effects on ACE (angiotensinconverting enzyme), is free from toxic effects on human umbilical vein endothelial cells (HUVEC), promotes releasing of endogenous relaxing factors in HUVEC and inhibits endothelin, thereby being capable of serving as antihypertensive peptide fragments with a good developing prospect for being applied to antihypertensive medicines, healthcare products and special medical foods.

A column-based device and method for retrieving cells of interest were enclosed. The said device comprises a column comprising (i) an inner wall defining an inner chamber with inlet and outlet openings, (ii) a perforated plug disposed adjacent to the outlet opening, (iii) a sleeve insert with a channel and disposed within the chamber and adjacent to the perforated plug, and (iv) a filtering means housed within sleeve insert sandwiched between two sealing means. In particular, Tumor-derived endothelial cell clusters (TECCs) as characterized multiple nuclei, expression of endothelial markers (PECAM1, VWF and CDH5), and non-expression of leukocyte, megakaryocyte and platelets markers, may be retrieved using the disclosed device. Also encompassed are methods, reagents and kits for the diagnosis and prognosis of cancers by detecting for the presence of TECCs isolated from blood samples using the claimed device.

The invention discloses a preparation for producing a COVID-19 antibody after oral administration and a preparation method of the preparation. Attenuated salmonella typhimurium is used as a carrier todeliver DNA vaccine, the preparation effectively reaches intestinal tract and delivers DNA vaccine to intestinal endothelial cells with low mammalian toxicity infection through oral enteric-coated capsules, bacteria is phagocytosed through macrophages, and specific molecules COVID-19-S is expressed to be recognized by immune cells, so that the immune effect of specific antibodies in mammals (mice, humans and other mammals) neutralizing COVID-19-S protein is achieved.

The invention belongs to the technical field of biomedicine, in particular to the technical field of an albumin medicament. In order to solve the technical problems of short half-life period in vivo and high use level of the existing ATWLPPR peptide, the invention provides an albumin variant. The albumin variant is obtained by mutating a KEQLK peptide section at the tail end of albumin C to a WLPPR peptide section. The albumin variant completely reserves the functions of human albumin and simultaneously has a new ATWLPPR sequence function. Experiments in vitro prove that the albumin variant can obviously inhibit the proliferation and migration of vascular endothelial cells induced by VEGF (Vascular Endothelial Growth Factor) in vitro, and the efficiency of the albumin variant corresponds to that of small peptide. Moreover, when the albumin variant is injected into animals, the efficiency of inhibiting the cornea angiogenesis functions of rabbits is 40-60 times higher than that of the small peptide with the same concentration. The albumin variant can be used as a novel anti-angiogenesis medicament with high efficiency.

The invention discloses an inhibitor for inhibiting the expression of MBD2 and application thereof, and relates to an inhibitor capable of inhibiting the expression of methylated CpG binding domain protein 2 in vivio and in vitro. The inhibitor comprises a nucleic acid molecule, a genetic structure, an siRNA molecule and a compound containing one or more of the nucleic acid molecule, the genetic structure and the siRNA molecule, and can be introduced into mammalian cells to inhibit the expression of the MBD2. The inhibitor for inhibiting the expression of the MBD2 is an siRNA, which has a nucleotide sequence in which the sense strand is 5'-GCAAGAGCGAUGUCUACUA-3' and the antisense strand is 5'-UAGUAGACAUCGCUCUUGC-3', or of which the 3' end is provided with two suspended basic groups tt, and has a nucleotide sequence in which the sense strand is 5'-GCAAGAGCGAUGUCUACUAtt-3' and the antisense strand is 5'-UAGUAGACAUCGCUCUUGCtt-3'. The application of the inhibitor in preventing and treating related diseases of vascular endothelial cell dysfunction provides a novel option for preparing medicaments or preparations for preventing and treating the related diseases of the vascular endothelial cell dysfunction by taking the MBD2 as a target.

The invention discloses a method for constructing a retinal neovascularization disease animal model by using a gene manipulation technology, a cultivation method and application, and relates to the technical field of gene editing. According to the construction method disclosed by the invention, the Ctnnnd1 gene in the vascular endothelial cells of the target animal is not expressed or the expression of the Ctnnd1 gene is inhibited by utilizing a gene manipulation technology. By means of the method, the animal model of the retinal neovascularization disease can be obtained, the variety of the animal model of the retinal neovascularization disease is enriched, more diversified model selections are provided for related research of the retinal neovascularization disease, and scientific research of the retinal neovascularization disease, development of related treatment drugs and the like are promoted.

An intracellular cytokine flow cytometry (CFC) assay was developed to measure CD3− (non-T) cell response to allo-Ags expressed on peripheral blood mononuclear cells (allo-CFC-PBMC) and / or endothelial cells (allo-CFC-EC) by detecting intracellular gamma-interferon (IFNY) production. The assay can be used to determine a likelihood of antibody mediated rejection in an individual. A method for using genetic screening to determine the likelihood of AMR is also disclosed.

The invention provides a p66Shc recombinant adenovirus vector. Homologous recombination in vitro of a human p66Shc fragment and a pAdeno X-CMV vector is realized by use of the gene engineering technology, and then competent bacteria are used for transformation to screen out correct recombinant adenoviruses; next, the recombinant adenoviruses are linearized and then used for transfecting HEK293A cells, and therefore, the recombinant adenoviruses are packaged and amplified largely in the HEK293A cells. After Hela cells are infected with the p66Shc recombinant adenovirus vectors, the growth and proliferation of the cells can be obviously inhibited; if human umbilical vein endothelial cells (HUVEC) are in infected with the p66Shc recombinant adenovirus vectors, no obvious growth inhibition effect is caused. The p66Shc recombinant adenovirus vector is expected to achieve a special growth inhibition effect on tumor cells.

The invention discloses a polypeptide and application thereof. The polypeptide contains continuous amino acid residue fragments with sequences of Pro-Gly-Ala-Asp or Asp-Ala-Gly-Pro. The polypeptide can be synthesized according to sequence structures by the aid of a polypeptide synthesizer and also can be extracted from natural plants including radix notoginseng, flos sophorae, aloe vera and\or rhizoma bletillae. The polypeptide and the application have the advantages that effects of promoting migration of stem cells and vascular progenitor cells of mice and directionally differentiating the stem cells and the vascular progenitor cells towards vascular endothelial cells can be realized by the polypeptide, and accordingly damage repair of the vascular endothelia cells and formation of new blood vessels can be promoted; skin spray can be prepared from the polypeptide, wound healing can be promoted by the skin spray, and inflammation can be prevented and treated by the skin spray; oral liquid for piglets can be prepared from the polypeptide, regeneration and repair of intestinal villi of the piglets can be promoted by the oral liquid, and accordingly diarrhea diseases of the piglets can be fundamentally prevented and radically treated by the oral liquid; feed additives for animals can be prepared from the polypeptide, and effects of resisting diarrhea and promoting growth can be realized by the feed additives; perfusion agents for the uteruses of sow can be prepared from the polypeptide, quick healing of postpartum wound of the sow can be promoted by the perfusion agents, and inflammation of the sow can be effectively prevented and treated by the perfusion agents.