143 results about "Endothelial Cell Growth Factor" patented technology

The invention concerns the prevention and treatment of endothelial injury and the injury of tissues containing injured blood vessels by administration of angiogenic factors, such as vascular endothelial cell growth factor (VEGF).

The invention discloses a preparation method of an essence containing human mesenchymal stem cell factors. The preparation method of the essence containing the human mesenchymal stem cell factors comprises the steps: preparation of freeze-dried powder of the human mesenchymal stem cell factors, preparation of a solvent and mixing. The preparation method obtains a high-purity and high-activity stem cell active factor concentrated solution by a cryoconcentration technique and the concentrated solution is freeze-dried to obtainfreeze-dried powder at low temperature in vacuum, so that the factor activity can be kept for a long time; and the solvent contains multiple skin nourishing and moisturizing components and a stem cell lysis solution, can well dissolve the cell factor freeze-dried powder and keeps the cell factor activity. The stem cell essence disclosed by the invention contains vascular endothelial cell growth factors, platelet derived growth factors, epidermal growth factors and other components, can promote skin regeneration, can play roles in moisturizing, tendering, whitening and repairing the skin, and has good application prospects in fields of medical beauty treatment, health protection and the like.

The present invention provides vascular endothelial cell growth factor (VEGF) antagonists and methods of using VEGF antagonists. VEGF antagonists contemplated by the invention include VEGF antibodies and VEGF receptor fusion proteins. Methods of treating edema and stroke using VEGF antagonists are also provided.

A mouse monoclonal antibody for antagonizing and inhibiting binding of a vascular endothelial cell growth factor (VEGF) and its receptor (VEGF-R), and a heavy chain variable region and light chain variable region amino acid sequence thereof. Also disclosed are a humanized preparation process of the antibody and a heavy chain variable region and light chain variable region amino acid sequence of the humanized antibody. The humanized antibody or its derivative can act as an ingredient of a pharmaceutical composition or be prepared into a suitable pharmaceutical preparation, is administered alone or in combination with a chemotherapy drug or other treatment means, and is used in broad-spectrum treatment of various solid tumors such as colon cancer, breast cancer and rhabdomyosarcoma.

The present invention relates to histoengineering bone comprising porous histoengineering bone rack material, composite heterogene seed cell and / or bioactive factor. The histoengineering bone is constituted through treating heterogene cancellous bone via hypotonic solution and ultrasonic cleaning, complete or partial decalcification, defatting, antigen eliminating, etc to obtain high porosity rack material; compounding mesenchyme stem cell, osteoblast and other seed cell and / or bone morphogonetic protein of the heterogene, vascular endothelial growth factor, antibacterial medicine and other bioactive factors; and applying human serum or no serum culture medium following culturing in ox serum culture medium to reduce heterologous serum residue so as to constitute histoengineering bone product ultimately. The histoengineering bone has high performance and is used as bone repairing material clinically.

The invention provides a human anti-vascular endothelial cell growth factor (VEGF) antibody as well as a coding gene and an application thereof. According to the invention, by virtue of genetic engineering means and a phage surface display technology, an anti-VEGF genetic engineering single-chain antibody is screened from a complete synthesis single-chain human antibody library, a variable region gene sequence of the anti-VEGF genetic engineering single-chain antibody is obtained, amino acids in a CDR (complementary determining region) of the anti-VEGF genetic engineering single-chain antibody are transformed by virtue of bioinformatics means, and finally a series of high-affinity anti-VEGF antibodies are obtained by constructing and screening mutation libraries and utilizing a chain exchange method, wherein affinity of the anti-VEGF antibodies with human VEGF is 10<-9>-10<-11>M. According to the invention, identification on immunocompetence and biological activity of the antibody is completed, the anti-VEGF antibody is verified to have the characteristic of being antagonistic to combination of VEGF and VEGFR2 and can effectively inhibit proliferation of vascular endothelial cells induced by VEGF. The invention provides a new specific candidate antibody molecule for resisting tumour and senile macular degeneration targeted by VEGF in future.

The disclosure provides a novel anti-angiogenesis fusion protein. The present invention combines a chimeric vascular endothelial cell growth factor (VEGF) receptor or a fragment thereof with a multimerizing component, which have a superior binding capacity with human VEGF and placental growth factor (PIGF). The fusion protein has improved stability, prolonged half-life and the ability to form multivalent interactions with VEGF, and can be used for anti-angiogenesis, treating VEGF related diseases and inhibiting tumor growth.

The present invention provides vascular endothelial cell growth factor (VEGF) antagonists and methods of using VEGF antagonists. VEGF antagonists contemplated by the invention include VEGF antibodies and VEGF receptor fusion proteins. Methods of treating edema and stroke using VEGF antagonists are also provided.

The present invention involves the preparation of vascular endothelial growth actor (VEGF) variants which provide materials that are selective in respect to binding characteristics to the kinase domain region and the FMS-like tyrosine-kinase region, respectively KDR and FLT-1. The respective KDR and FLT-1 receptors are bound by corresponding domains within the VEGF compound domains. The variants hereof define those two binding regions and modify them so as to introduce changes that interrupt the binding to the respective domain. In this fashion the final biological characteristics of the VEGF molecule are selectively modified.

The invention discloses a stem cell culture medium. The stem cell culture medium comprises an improved DMEM (Dulbecco's Modified Eagle Medium) / F12 basal culture medium and an additive, wherein the additive comprises sodium bicarbonate, selenium amino acid chelate, recombinant human insulin growth factors, recombinant human basic fibroblast growth factors, recombinant human lactoferrin, ascorbic acid, recombinant human platelet-derived growth factors, recombinant human vascular endothelial cell growth factors, octacosanol, polyvinyl alcohol, polyvinylpyrrolidone and recombinant human epidermal growth factors. According to the stem cell culture medium, the potential of stem cells is not influenced while the stem cells can be proliferated rapidly, the proliferation speed of the stem cells is increased by 3-5 times compared with a common culture medium, and further, the stem cell culture medium can be used for culturing the stem cells of various kinds of tissue and has excellent applicability; the cultured stem cells have high differentiation capability, can be differentiated into multiple functional cells and have very high scientific research and medical application values, culture medium components are exact, the quality is stable, and accordingly, the cultured stem cells are not likely to generate human body rejection reaction after transplanting.

The invention belongs to the technical field of genetic engineering antibodies and particularly relates to a whole human source single chain antibody which is designed and expressed by genetic engineering and has specific affinity towards human vascular endothelial growth factor receptor 2 (VEGFR-2). The invention also provides a nucleotide sequence of heavy chain variable region and light chain variable region immunoglobulin molecules, a nucleotide sequence which comprises the heavy chain variable region and light chain variable region immunoglobulin molecules, an amino acid sequence which comprises variable heavy chain and variable light chain immunoglobulin molecules, and a sequence which corresponds to CDR1, CDR2 and CDR3 of a complementary determining region. The invention also provides a method for generating and expressing the anti-VEGFR-2 single chain antibody, by cycling operations of gathering-elution-gathering, an antibody which has specific affinity towards the human vascular endothelial growth factor is screened, and then the single chain antibody is obtained through a prokaryotic cell secretion and expression system and an affinity purification system. The antibody can be coupled with a detectable substance and therapeutic agent.

The invention belongs to the field of biotechnology-monoclonal antibody. The present invention discloses a monoclonal antibody for antagonistic inhibition of combination of vascular endothelial cell growth factor (VEGF) and a receptor of the VEGF, and a hybridoma cell line secreting the monoclonal antibody, wherein the preservation number of the hybridoma cell line is CGMCC No.5889. The present invention further discloses a preparation method and a use of the monoclonal antibody, wherein the use comprises application of the monoclonal antibody in VEGF protein detection and application of the monoclonal antibody in preparation of a drug preparation for treatment of angiogenesis-related diseases.

The invention discloses a preparation method of a surface coating of intravascular stents or cardiac valves with excellent biocompatibility, which includes the steps: A. the preparation of a coating solvent: heparin is dissolved in deionized water, the pH value thereof is adjusted to 7 to 8 and then an endothelial cell growth factor is added to form a heparin / growth factor solution; collagens are dissolved in acetum to form a collagen solution; B. dip-coating: the intravascular stents or the cardiac valves are immersed in the heparin / growth factor solution and a layer of heparin / growth factor is dipped and coated on the surface of the intravascular stents or the cardiac valves, and then the intravascular stents or the cardiac valves are taken out for being cleaned by the deionized water and being dried by nitrogen gas; and the intravascular stents or the cardiac valves are immersed in the collagen solution, coated with a layer of collagen, and taken out for being cleaned by the deionized water and being dried by nitrogen gas; the steps of dip-coating by the use of the heparin / growth factor and the collagen are repeated in sequence for 1 time to 60 times; finally the step of dip-coating by the use of the heparin / growth factor is carried out again to obtain the surface coating with excellent biocompatibility of the intravascular stents or the cardiac valves. The intravascular stents or the cardiac valves prepared have excellent biocompatibility and low preparation cost.

A novel growth factor specific for vascular endothelial cells has been identified in conditioned medium of bovine pituitary derived folliculo stellate cells. This factor, named folliculo stellate derived growth facto (FSdGF) or vascular endothelial growth factor (VEGF), was purified to homogeneity by a combination of heparin sepharose affinity chromatography, Bio Gel P-60 exclusion chromatography, Mono S ion exchange chromatography and hydrophobic chromatography on a C4 reverse phase HPLC column. The factor is also found in the murine AtT-20 cell line.

A novel growth factor specific for vascular endothelial cells has been identified in conditioned medium of bovine pituitary derived folliculo stellate cells. This factor, named folliculo stellate derived growth facto (FSdGF) or vascular endothelial growth factor (VEGF), was purified to homogeneity by a combination of heparin sepharose affinity chromatography, Bio Gel P-60 exclusion chromatography, Mono S ion exchange chromatography and hydrophobic chromatography on a C4 reverse phase HPLC column. The factor is also found in the murine AtT-20 cell line. Alternatively, the growth factor is purified by a first reverse phase HPLC using acetonitrile gradient followed by a second reverse phase HPLC using an isopropanol gradient. FSdGF, having a molecular weight of about 43,000 da, was characterized as a glycoprotein composed of two homologous sub units with MW of about 23 kDa. FSdGF was a potent mitogen for vascular endothelial cells with activity detectable at 10 pg / ml and saturation at 500 pg / ml. It did not stimulate the proliferation of other cell types such as bovine corneal endothelial cells, adrenal cortex cells, granulosa cells, BALB / MK cells or BHK-21 cells. Microsequencing revealed an amino terminal sequence containing no significant homology to any known protein. The release of FSdGF by pituitary cells and its unique target cell specificity indicate that FSdGF is useful in angiogenesis.

The invention discloses a fusion protein capable of inducing and activating cancer targeting T-cells as well as a preparation method and application of the fusion protein. The fusion protein contains peptide acting with cancer cells as well as costimulatory molecules B7.1, wherein the peptide acting with the cancer cells is selected from a transforming growth factor-alpha, an epidermal growth factor, a vascular endothelial growth factor, gonadotropinreleasing hormone or gastrin-releasing peptide. The fusion protein has a targeting function; the fusion protein can act with VEGFR, EGFR, GnRH-R, or GRP-R respectively on one hand and interact with corresponding receptors CD28 and CTLA-4 expressed on the T-cells, therefore the targeting of the T-cells is positioned at the periphery of the cancer cells greatly expressing VEGFR, EGFR, GnRH-R, or GRP-R. Proven by experiments, the fusion protein can restrain the growth of tumors and cause cancer cells to apoptosis.

The invention provides a double-targeted fusion protein of target programmed death-1 (PD-1) or target programmed death-1 ligand (PD-L1) and a target vascular endothelial cell growth factor (VEGF) family. The double-targeted fusion protein comprises (i) an anti-PD-1 antibody or an anti-PD-L1 antibody and (ii) a VEGF family inhibiting domain (VID) effectively connected to the C end of each of two heavy chains of the anti-PD-1 antibody or the anti-PD-L1 antibody. The invention further provides polynucleotide coding the double-targeted fusion protein, a carrier containing the polynucleotide, a host cell containing the polynucleotide or the carrier, and application of the double-targeted fusion protein to therapy, prevention and / or diagnosis of diseases related to PD-1 activity, PD-L1 activityand VEGF family activity in an individual.

The invention discloses an inhibition method for induced differentiation of hair follicle stem cells into vascular endothelial cells, comprising: (1), isolating rat hair follicle stem cell; (2), culturing the rat hair follicle stem cells; (3), purifying the rat hair follicle stem cells; (4), inhibiting the induced differentiation of the rat hair follicle stem cells into vascular endothelial cells. In the inhibition method provided herein, vascular endothelial cell growth factor 165 is used for the first time as an inducing factor so that the rat hair follicle stem cells efficiently and directionally differentiate into vascular endothelial cells; the formation and growth of vascular endothelial cells generated by induced differentiation of the hair follicle stem cells can be adjusted, an effective healing of a wound is promoted; it is also possible to provide seed cell sources for solving the problems in vascularization of tissue engineering skin, cell-transplant therapy of ischemic disease and the like.

The invention relates to a method for preparing an in-situ tissue engineering blood vessel by a composite process. According to the invention, a three-layer composite intravascular stent which is similar to a natural vascular structure and is used for in-situ tissue engineering is prepared by combining rapid prototyping, electrostatic spinning and wet spinning processes; wherein the inner layer isa tubular stent provided with an axial groove by utilizing 3D printing and electrostatic spinning technologies; a middle layer is a tubular stent which is prepared by wet spinning and is provided with circumferentially arranged fibers; and the outer layer is the tubular stent which is prepared by the coatable inner layer obtained by electrostatic spinning and the disordered fibers at the middle layer. Through heparinization, the vascular endothelial cell growth factor is loaded, so that cells can be collected in a host, the infection opportunity of in-vitro culture is reduced, the preparationof in-situ tissue engineering blood vessels is realized, and the method has a wide prospect in the field of tissue engineering blood vessels.

The invention discloses a nerve repairing material and a preparation method thereof. The nerve repairing material comprises a combination of cell factors capable of promoting nerve growth, and a controlled-release carrier of the cell factors, wherein the carrier is mainly composed of fibrinogen, fibronectin, heparin, fibrin stabilizing factor, thrombin and calcium chloride, wherein the combination of the cell factors capable of promoting nerve growth is optimally at least two of nerve growth factor, brain-derived neurotrophic factor, basic fibroblast growth factor and vascular endothelial cell growth factor, and is embedded in the carrier. According to the nerve repairing material, the speed of releasing cell factors can be regulated according to the nerve repairing progress by utilizing 'intelligent release' of the controlled-release carrier, so that the functions of cell factors can be progressively played; furthermore, the nerve repairing can be further promoted by utilizing the synergy of the cell factors.

A novel growth factor specific for vascular endothelial cells has been identified in conditioned medium of bovine pituitary derived folliculo stellate cells. This factor, named folliculo stellate derived growth facto (FSdGF) or vascular endothelial growth factor (VEGF), was purified to homogeneity by a combination of heparin sepharose affinity chromatography, Bio Gel P-60 exclusion chromatography, Mono S ion exchange chromatography and hydrophobic chromatography on a C4 reverse phase HPLC column. The factor is also found in the murine AtT-20 cell line. Alternatively, the growth factor is purified by a first reverse phase HPLC using acetonitrile gradient followed by a second reverse phase HPLC using an isopropanol gradient. FSdGF, having a molecular weight of about 43,000 da, was characterized as a glycoprotein composed of two homologous sub units with MW of about 23 kDa. FSdGF was a potent mitogen for vascular endothelial cells with activity detectable at 10 pg / ml and saturation at 500 pg / ml. It did not stimulate the proliferation of other cell types such as bovine corneal endothelial cells, adrenal cortex cells, granulosa cells, BALB / MK cells or BHK-21 cells. Microsequencing revealed an amino terminal sequence containing no significant homology to any known protein. The release of FSdGF by pituitary cells and its unique target cell specificity indicate that FSdGF is useful in angiogenesis.

The invention discloses a mimic short peptide 7B of endothelial cell growth factor VEGF antigen epitope and an application thereof. The amino acid sequence of the mimic short peptide 7B is FKPSCVPLMRCGGCCN and the nucleotide sequence is TTCAAACCGTCCTGCGTTCCGCTGATGCGTTGCGGTGGTTGTGCAACG. In vitro experiments show that the mimic short peptide 7B of the invention has obvious inhibiting effect during the processes of endothelial cell proliferation and tube formation and the tumor cell proliferation and migration. Therefore, the mimic short peptide 7B of the invention can be used for preparing peptide vaccines, tumor angiogenesis inhibitor or tumor-oriented drugs, and has great reference value on diagnosing and predicting tumorigenesis by utilizing VEGF receptor as a target, discussing development of small molecular peptides on the tumor angiogenesis inhibitors, and developing and researching tumor-oriented drugs.

The present invention provides VEGF variants having one or more amino acid mutations in the KDR and / or FLT-1 receptor binding domains in the native VEGF sequence and selective binding affinity for either the KDR receptor or the FLT-1 receptor. Methods of making the VEGF variants and methods of using the VEGF variants are also provided.

The invention belongs to the field of biotechnology and provides a stem cell induced culture solution used for eyes .The stem cell induced culture solution comprises a vascular endothelial cell growth factor (VEGF), a basic fibroblast growth factor (bFGF) and an epidermal growth factor (EGF); the content of the vascular endothelial cell growth factor (VEGF) is 0.5-2.5 ng / ml, the content of the basic fibroblast growth factor (bFGF) is 0.25-0.4 ng / ml, and the content of the epidermal growth factor (EGF) is 0.5-2.0 ng / ml .The stem cell (ASCs) in vitro conditioned culture medium from human eyepit fat is adopted, and a significant reinforcement function is achieved for regeneration and repair of corneal injury.

The invention discloses an individualized 3D (three-dimensional) printed bone tissue engineering stent with a mosaic structure. The stent is characterized in that a shape and a structure of a large area of bone defect of a jaw are rebuilt through 3D modeling. The stent comprises two parts, wherein one part is an outer outline stent used for restoring the shape and the structure of a cranio-facial bone defect, and the other part is an inner tubular stent used for promoting formation of blood vessels. The bone tissue engineering stent with the maintained shape and structure on the outer part and promoted formation of blood vessels in the inner part is formed by inoculating factors for promoting ossification, such as mesenchymal stem cells (MSCs) and / or a loaded bone morphogenetic protein (BMP), onto the outer stent, and inoculating factors for promoting the formation of the blood vessels, such as endothelial cells (ECs) and / or a loaded vascular endothelial cell growth factor (VEGF), onto the inner stent, so that stem cell proliferation, ossification differentiation, and angiopoiesis of a tissue engineering compound are facilitated, and quality and speed of osteanagenesis are improved.

The invention discloses a culture medium for rapid proliferation of neural stem cells. The culture medium is prepared from an RPMI-1640 culture medium, trehalose, recombinant human insulin, insulin growth factors, endothelial cell growth factors, potassium chloride, amino acid chelated selenium, glutamine, progesterone, coenzyme I, lipoic acid, vitamin, polygahatous polysaccharides, polysaccharide from radix codonopsis and ginseng saponin. The culture medium for rapid proliferation of the neural stem cells is good in cell wall adherence, fast in proliferation and free of animal source ingredients and does not produce residues or pollution, meanwhile the anti-pathogen microbial effect of the culture medium can be improved, and the cost of the culture medium can be reduced.

This invention relates to a fusion protein that possess action of selectively kill tumour rebirth blood vessel endotheliocyte, and its application. This fusion protein through connecting peptide connect VEGF121 with amido end of sponge gourd seed ribosome inactivating protein(RIP) Luffin Alpha. The vascular endothelial cell growth factor VEGF121 act as means of delivery, make this fusion toxin idiosyncratic incorporate with vascular endothelial cell growth factor receptor F1k1 / KDR, thereby optionally ingress tumour rebirth vascular endothelial cell; the polypeptide possess amphipathic molecule character, by destroy lysosome membrane to promote the release of lysosome inside dissociate toxin molecule; the toxin carboxyl terminal endoplasmic reticulum loacting signal led toxin molecule( sponge gourd seed RIP Luffin Alpha) to target site to exert toxic effect, so to destroy rebirth blood vessel of tumour organization, cut off tumour blood supply, achieve the end of restraining tumor.

The invention discloses a cryoprotectant for umbilical cord mesenchymal stem cells and application of the cryoprotectant. The cryoprotectant comprises a human serum albumin solution, a dimethyl sulfoxide solution, trophic factors and a pharmaceutically-acceptable carrier, and the trophic factors are one or more of a vascular endothelial cell growth factor VEGF, a transforming growth factor TGF, interleukin 6IL-6, a fiber bud cell growth factor bFGF and a hepatocyte growth factor HGF. The invention discloses the application of the cryoprotectant for the umbilical cord mesenchymal stem cells incryopreservation of the umbilical cord mesenchymal stem cells. Through the above mode, according to the application, the specific cryoprotectant is adopted to cryopreserve the umbilical cord mesenchymal stem cells, allows the cryopreserved umbilical cord mesenchymal stem cells to keep activity and differentiation stemness, and has a good cryopreservation effect.

The invention relates to a stem cell combined growth factor injection for promoting angiogenesis in ischemic tissue, a preparation method and a use method thereof. The body’s protective response to peripheral tissue ischemia is relatively complex. Although a single growth factor intervention is effective, it cannot complete this process; although stem and progenitor cell transplantation can improve the vascular density and blood perfusion of ischemic tissue, the main source of cells is bone marrow. There are serious limitations in the acquisition of umbilical cord blood, umbilical cord blood, etc., and the in vivo viability of cells is extremely limited, which seriously affects its therapeutic effect. The invention utilizes human basic fibroblast growth factor to intervene in adipose mesenchymal stromal cells, and cooperates with vascular endothelial cell growth factor to promote angiogenesis in ischemic tissue. The present invention overcomes the benefit limitation of single factor or cell therapy, ensures sufficient stem cell acquisition, and improves the survival ability and angiogenesis effect of transplanted cells, thereby further improving blood perfusion of ischemic lower limbs and improving limb survival Rate.