43 results about "Murine fibroblast" patented technology

We have made retrovirus particles displaying a functional antibody fragment. We fused the gene encoding an antibody fragment directed against a hapten with that encoding the viral envelope protein (Pr80env) of the ecotropic Moloney murine leukemia virus. The fusion gene was co-expressed in ecotropic retroviral packaging cells with a retroviral plasmid carrying the neomycin phosphotransferase gene (neo), and retroviral particles with specific hapten biding activities were recovered. Furthermore the hapten-binding particles were able to transfer the neo gene and the antibody-envelope fusion gene to mouse fibroblasts. In principle, the display of antibody fragments on the surface of recombinant retroviral particles could be used to target virus to cells for gene delivery, or to retain the virus in target tissues, or for the construction of libraries of viral display packages.

The invention discloses a method for preparing a nanosilver / nano silicon dioxide-containing antibacterial biological dressing, and relates to the field of antibacterial biological dressings. The nanosilver / nano silicon dioxide-containing antibacterial biological dressing is prepared by adding mixed solution of chitosan and polyvinyl alcohol and silver-carried silicon dioxide powder during the acetalation of the polyvinyl alcohol. The nanosilver / nano silicon dioxide-containing antibacterial biological dressing of the invention can accelerate the wound healing, slowly release the silver nanoparticles and continuously act on reproductive bacteria, has the characteristic of durable and high-efficiency sterilization and does not produce medicament resistance; and through the researches on the toxicity of the material to mouse fibroblast cells, the product does not produce obvious toxicity to the cells but promotes the growth of the cells.

The invention relates to a polyurethane material with protein adsorption resistance and cell adhesion resistance and a preparing method thereof. The material comprises a substrate and a modification layer. The substrate is made of polyurethane. The modification layer is obtained by polymerizing a vinyl pyrrolidone monomer on the surface of the substrate by utilization of an atom transfer radical polymerization technology. The material is prepared by steps of: subjecting the polyurethane to isocyanic acid esterification, performing amination, fixing a bromine initiator, and initiating graft polymerization of the vinyl pyrrolidone on the surface of the material. The surface of the polyurethane material prepared by the method has good hydrophilicity, good resistance to bovine serum fibrinogen adsorption, bovine serum albumin adsorption and lysozyme adsorption, and a function of resisting cell adhesion of mouse fibroblast cells (L-929), so that the polyurethane material is suitable for preparing biomedical catheter materials with complex shape structures.

The invention discloses a preparation method of a chitosan / polyvinyl alcohol sponge dressing containing nano-silver. The invention relates to an antibacterial dressing for therapy of trauma, and the chitosan / polyvinyl alcohol sponge dressing containing the nano-silver is prepared by adding nano-silver / polyvinyl alcohol solution and polyvinyl alcohol / chitosan blend solution. The obtained dressing has good biocompatibility and hemostatic and antibacterial effects, and can promote the wound healing; the product can accelerate the wound healing, release nano-silver particles in a sustained manner, be acted on regenerated bacteria constantly, have the lasting and high-efficient sterilization characteristics and avoid the drug resistance during the wound treatment process; the research on the toxic effect of the material to mouse fibroblasts shows that the product has no obvious toxicity to the cells and has the effect of promoting the growth of the cells; and preparation method has simple process equipment and low price of experimental raw materials and is conductive to industrialization.

The invention discloses a preparation method of chitosan / acetalized poval medical dressing, and in particular relates to a preparation technology of poval sponge. A dressing with bioactivity can be prepared by adding a poval / chitosan mixed solution in poval acetalation reaction. The chitosan / acetalized poval medical dressing has biocompatibility,the effects of inflammation, blood stopping, wound surface healing promotion, good breathability, water absorbability and tensile strength; in addition, proved by the search on toxic effect on mouse fibroblast cells, the chitosan / acetalized poval medical dressing has no obvious toxicity to the mouse fibroblast cells, but has the effect of promoting the growth of the mouse fibroblast cells.

This invention uses mesoporous silica nanoparticles and other nanostructured materials to formulate polyacrylate-based bone cement for achieving an enhanced and controlled elution of active ingredients such as antibiotics. This invention overcomes the limitation of low antibiotic release from commercial polyacrylate-based bone cements using for example, PMMA. In certain aspects, the formulation enables a sustained release of antibiotics from the bone cement over a period of 80 days and achieves 70% of total drug release, whereas the commercial antibiotic bone cement (e.g., SmartSet GHV) only releases about 5% of the antibiotics on the first day and subsequently an almost negligible amount. In addition, the mechanical properties of our formulated bone cements are well retained. The inventive bone cement exhibits good antibacterial properties and has very low cytotoxicity to mouse fibroblast cells.

The invention provides a sulfonic quantum dot / silk fibroin composite nanofiber membrane and a preparation method and application thereof. The preparation method comprises the following steps that degumming treatment is performed on natural silk, and silk fibroin is obtained; then, fibroin is dissolved in a lithium bromide water solution, and a silk fibroin sponge is obtained through dialysis, centrifugal and freeze drying; the silk fibroin sponge is dissolved into formic acid to be mixed to be uniform, then, a sulfonate graphene quantum dot water solution is ultrasonically and uniformly dispersed into a silk fibroin-formic acid solution, and a composite nanofiber membrane is prepared through electrostatic spinning equipment; mouse fibroblasts are used for performing cell viability evaluation on the composite nanometer member, and it shows that the composite nanometer membrane has the excellent biocompatibility. The prepared composite fiber membrane has the good biocompatibility and degradability, has the potential for promoting proliferation and differentiation of human bone marrow storm stem cells, and has the good biologic medical material application prospect.

The invention discloses a method for inducing mouse fibroblasts into cartilage by adopting a small-molecule composition. The method comprises the following steps: carrying out adherent culture on mouse embryo fibroblasts, removing a culture medium, slowly adding a chemical induction culture medium containing the small-molecule composition, carrying out culturing in the environment having the temperature of 37 DEG C and containing 3-8% of oxygen, 3-8 % of carbon dioxide and the balance of nitrogen, wherein the chemical induction culture medium containing the small-molecule composition is replaced every 2-3 days; carrying out continuous culturing for 4-12 days, thus obtaining intermediate state cells, wherein the small-molecule composition comprises an HDAC inhibitor, a GSK-3 inhibitor and aTGF-beta signal channel inhibitor; and transferring the intermediate state cells to a cartilage inducing medium, carrying out culturing in the environment having the temperature of 37 DEG C and containing 15-25% of oxygen, 3-8 % of carbon dioxide and the balance of nitrogen, wherein the cartilage inducing medium is replaced by the fresh cartilage inducing medium once every 3-4 days, and carryingout culturing for 14-28 days, thus obtaining a cartilage cell cluster. With the method provided by the invention, the problem that in the traditional methods, the fibroblasts can be induced to form cartilage cells through transdifferentiation only after an exogenous gene is introduced is solved, and the method provided by the invention is expected to be used for further solving the problem that seed cells of cartilage cells are in shortage or in-situ focus fibrosis exists.

The invention discloses a mouse fibroblast and an application thereof, and belongs to the field of cell biology. The cell is named mouse fibroblast MFC / HL-041 with the preservation number of CCTCC NO:C201714. The mouse fibroblast treated by mitomycin C or irradiated by radioactive rays is used for separation and subculture of normal primary epithelial cells and primary tumor cells. Obtained humanpulmonary normal primary epithelial cells and lung cancer primary epithelial cells are fresh and live in state, tight in arrangement, clear in cell boundary, high in stereoscopic impression and polygonal when examined under a microscope. The mouse fibroblast can be used for physiological study of human or animal normal cells, a virus infection model of in-vitro normal cells, drug sensitivity tests of the in-vitro normal cells for different drugs, drug screening for individualized treatment of cancer patients as well as research and development of new anti-cancer drugs.

The invention discloses a capsid protein of an adeno-associated virus (AAV) which has an amino acid sequence shown as SEQ ID NO: 1. The invention also discloses an isolated nucleic acid molecule whichencodes the capsid protein of the AAV. The invention further discloses a vector of the AAV which encodes the capsid protein of the AAV. The invention still further discloses a construction method andapplication of the vector of the AAV. According to the capsid protein of the AAV, modification is performed on the capsid protein of the AAV-DJ , so that the AAV-DJ has significantly improved transduction efficiency in cell lines, including HEK293, various liver cancer cells, mouse fibroblasts and the like, and mouse livers compared with wild-type AAV, so that immune response in subjects caused by high-dose injection of virus vectors is avoided; and moreover, production process of the AAV is optimized so as to reduce production cost. Thus, clinical progress of AAV gene therapy products is promoted

A method is disclosed for measuring the hydrogen and / or oxygen isotope ration of intracellular water in Escherichia coli cells and correlating the hydrogen and / or oxygen isotope ratio with the metabolic activity of the cell. A method is also disclosed for measuring the hydrogen and / or oxygen ratio of intracellular water via a probe, such as a fatty acid, and correlating the hydrogen and / or oxygen ratio with the metabolic activity of the cell. Methods for measuring the hydrogen and / or oxygen isotope ratio of water from eukaryotic organisms, such as cultured rat fibroblasts and whole mammals, and optionally relating the same to a metabolic rate, are also disclosed.

The present invention discloses a method for preparing induced pluripotent stem cells by using a Xist Tale inhibitory transcription factor. Synergism of the transcriptional activator-like effectors (Tale)-based inhibitory transcription factor combined to a first intron region of Xist, Oct4, Sox2, Klf4 and c-Myc enhances the efficiency of induction of mouse fibroblasts to the induced pluripotent stem cells. Obtained iPSCs have typical pluripotent stem cell characteristics including involvement in the development of chimeric mice. The method actively promotes the clinical application and researches of the iPSCs, and provides new technical and theoretical bases for studying how to obtain and improve the iPSCs in people, pigs, cattle and other mammals.

The invention discloses a tissue engineering lymph node model and a construction method thereof and belongs to the technical field of tissue engineering and regenerative medicine. The tissue engineering lymph node model comprises lymphocytes separated from lymphoid tissues and a collagen-Matrigel scaffold. The construction method for the tissue engineering lymph node model comprises the following steps of: preparing lymphocyte suspension and primary generation mouse fibroblast cell suspension; and compounding mixed cell suspension and the collagen-Matrigel scaffold, and culturing in a mold subjected to static stretching to obtain the tissue engineering lymph node model. The scaffold system subjected to static stretching can better simulate in-vivo environment and support the growth and proliferation of the lymphocytes, secretion of cell factors and the expression of surface markers, and has important significance for applying the tissue engineering lymph node model to immunotoxicity evaluation of biological materials, immune medicine screening and the like. The construction method has the advantages of simple operating process and mild implementation conditions.

The invention discloses a preparation method of antibacterial silicone rubber. The preparation method comprises the following step: inducing ring opening polymerization of amino acid N-carboxylic acid anhydride in situ by virtue of amino on the surface of silicone rubber, so as to obtain the silicone rubber with the surface containing a polyamino acid copolymer or a polyamino acid homopolymer. According to the antibacterial silicone rubber prepared by virtue of the preparation method, the problems that existing antibacterial silicone rubber easily generates drug resistance and is poor in biocompatibility are solved, and the antibacterial silicone rubber has good application prospects. The antibacterial rate of the prepared antibacterial silicone rubber to bacteria including escherichia coli, staphylococcus aureus, staphylococcus epidermidis, pseudomonas aeruginosa, Candida albicans, pneumococcus and the like exceeds 90%, and the cytotoxicity of mouse fibroblast does not exceed level 1.

Disclosed is a simple method of determining SPF based on the staining pattern of viable cells like mouse fibroblasts and human keratinocytes after exposure to ultraviolet radiations. The method provides results comparable to the standard methods of SPF measurement using human subjects.

The present invention provides a new method for gene point mutation repair. Main steps are as follows: in a target genome sequence, cell gene mutation sites are selected as repaired gene sites; according to the selected gene repair sites, sgRNA of a CRISPR / Cas 9 system is designed; the sgRNA is constructed to an expression vector carrying CRISPR / Cas9 proteins; a segment of correct sequence is selected near upstream or downstream of the mutation sites, and a designed Donor DNA is used as a template for the gene mutation repair; in vitro culture of cells carrying the mutation site genes is conducted; an electrotransfection method is used to transfect the sgRNA, CRISPR / Cas9 expression vector and Donor DNA into the cells. The method fully mixes a Cas9 expression vector, the sgRNA and the DonorDNA with mouse fibroblast cell line carrying Msh5 point mutation established in vitro, then the cell line is electrically transfected to increase transfection efficiency of the cells, and the CRISPR / Cas9 system is used to conduct the repair of the mutation sites, thereby providing a scientific basis for treatment of primary amenorrhea in women.

The present invention includes a method for preparing a three-dimensionally cultured skin model comprising dermis and epidermis, which comprises: a step of preparing the dermis using a composition comprising murine fibroblasts; and a combination of native collagen and atelocollagen; and a step of forming the epidermis using keratinocytes. Also, the present invention relates to a three-dimensionally cultured skin model which comprises: a dermis prepared by a composition comprising murine fibroblasts, native collagen, or a combination of native collagen and atelocollagen; and epidermis formed from keratinocytes.

The invention discloses a method for promoting reprogramming of mouse fibroblasts into myocardial cells under low oxygen conditions. A reprogramming factor is utilized to directly reprogram mouse fibroblasts into efficiently differentiated myocardial cells; on such basis, the efficiency of low oxygen on direct reprogramming to generate myocardial cells and the variation of the HIF-1 expression level in the reprogramming process from fibroblasts to myocardial cells are detected, and the effect of low oxygen on the cell direct reprogramming is systematically explained. The expression conditions of the multi-potential transcription factor and epigenetic regulation factor under low oxygen stimulation are detected. The individualized myocardial cells obtained by directly reprogramming body cells are used for repairing myocardial infarction, thereby laying a foundation for enhancing the clinical cellular transplantation efficiency and safety. The research of the relationship between the low oxygen microenvironment and reprogramming provides a new idea for researching the reprogramming mechanism under low oxygen conditions.

The invention specifically relates to a chloroquine cholesterol derivative, and a preparation method and application thereof, belonging to the field of the chemical pharmaceutical industry. The chloroquine cholesterol derivative provided by invention has a structure as shown in a formula I which is described in the specification. The invention also provides the preparation method and application of the chloroquine cholesterol derivative. The chloroquine cholesterol derivative provided by the invention can effectively inhibit the proliferation of rat fibroblasts and treat pulmonary fibrosis, exerts anti-inflammatory effect, and offers a novel option for preparation of anti-fibrotic, anti-inflammatory and anti-tumor drugs.

The invention discloses a mouse fibroblast tumor cell strain HXLyAF-KBM and application thereof.The cell strain is derived from the bone marrow microenvironment in a human lymphoma cell nude mouse transplantation tumor model, the cell name of the cell strain is mouse lymphoma related fibroblast tumor cells HXLyAF-KBM, the preservation number is CCTCC NO: C2021106, the preservation date is December 9, 2021, and the preservation number is CCTCC NO: C2021106. The preservation unit is China Center for Type Culture Collection. The cell strain is a bone marrow-derived lymphoma-related mouse fibroblast tumor cell strain established for the first time at home and abroad, and can be applied to construction of a mouse fibroblast tumor animal model, a malignant bone marrow microenvironment model, a 3D tumor culture system and organoid system model, and a bone marrow microenvironment and lymphoma progress and drug resistance relationship model.

The invention discloses Tibetan ganoderma lucidum polysaccharide GLP-1 with antioxidant efficacy, a preparation method and application; wherein the ganoderma lucidum polysaccharide GLP-1 is extractedfrom Tibetan ganoderma lucidum powder, the polysaccharide content is 73.36%, the uronic acid content is 2.27%, the weight-average molecular weight is 6.31 kDa, and the protein content is extremely low. The Tibetan ganoderma lucidum polysaccharide GLP-1 is separated and prepared from Tibetan ganoderma lucidum for the first time, wherein the GLP-1 comprises mannose, glucose, galactose, xylose, arabinose and the like, has characteristic absorption peaks of OH, CH, COH, alpha isomeric pyranose and alpha, beta glucosidic bonds, does not have a triple helix structure, and is good in thermal stability; besides, the obtained polysaccharide GLP-1 has chemical antioxidant activity, can protect mouse fibroblasts NIH3T3 from oxidative damage caused by tert-butyl hydroperoxide induction, and has good edible and medicinal values in the aspect of antioxidant activity.

The invention discloses a method for preparing a nanosilver / nano silicon dioxide-containing antibacterial biological dressing, and relates to the field of antibacterial biological dressings. The nanosilver / nano silicon dioxide-containing antibacterial biological dressing is prepared by adding mixed solution of chitosan and polyvinyl alcohol and silver-carried silicon dioxide powder during the acetalation of the polyvinyl alcohol. The nanosilver / nano silicon dioxide-containing antibacterial biological dressing of the invention can accelerate the wound healing, slowly release the silver nanoparticles and continuously act on reproductive bacteria, has the characteristic of durable and high-efficiency sterilization and does not produce medicament resistance; and through the researches on the toxicity of the material to mouse fibroblast cells, the product does not produce obvious toxicity to the cells but promotes the growth of the cells.

The present invention provides a method for preparing retinal ganglion cells, which comprises the following steps of: obtaining a somatic cell; constructing lentiviral particles, transfecting the somatic cell with the lentiviral particles, integrating an Ascl1 gene, a Islet1 gene, and a Brn3b gene into the somatic cell chromosome; and performing induction culture to obtain the retinal ganglion cells. The retinal ganglion cells are obtained by integrating the exogenous Ascl1 gene, the Islet1 gene, and the Brn3b gene into the mouse fibroblast chromosome for overexpression preparation. Compared with the prior art, the present invention creatively selects a combination of the Ascl1 gene, the Brn3b gene and the Islet1 gene, the three transcription factors are overexpressed in the somatic cellsby lentivirus-mediated means, and more than 10% of the somatic cells can be transdifferentiated into Brn3a-positive, Tuj1-positive retinal ganglion cells in 7 days of induction culture, and the efficiency is high.

The invention discloses a preparation method of a chitosan / polyvinyl alcohol sponge dressing containing nano-silver. The invention relates to an antibacterial dressing for therapy of trauma, and the chitosan / polyvinyl alcohol sponge dressing containing the nano-silver is prepared by adding nano-silver / polyvinyl alcohol solution and polyvinyl alcohol / chitosan blend solution. The obtained dressing has good biocompatibility and hemostatic and antibacterial effects, and can promote the wound healing; the product can accelerate the wound healing, release nano-silver particles in a sustained manner, be acted on regenerated bacteria constantly, have the lasting and high-efficient sterilization characteristics and avoid the drug resistance during the wound treatment process; the research on the toxic effect of the material to mouse fibroblasts shows that the product has no obvious toxicity to the cells and has the effect of promoting the growth of the cells; and preparation method has simple process equipment and low price of experimental raw materials and is conductive to industrialization.

The invention provides feeder-layer cells and a preparation method thereof. The preparation method comprises the following steps: (1) cell culture: inoculating a culture dish with fibroblasts of a mouse, adding a culture solution, carrying out culture, and stopping culture when cell density reaches 80 to 90% so as to obtain cultured cells; (2) treatment with methanol: taking out the culture dish, removing the culture solution, cleaning the cultured cells with a phosphate buffer, then removing the phosphate buffer, adding methanol, carrying out standing at room temperature for 5 min, then removing methanol, carrying out standing on an ultra-clean workbench for 3 to 6 min and sealing the culture dish with a sealing membrane so as to obtain the feeder-layer cells. The invention also provides application of the feeder-layer cells to culture of multi-potent stem cells. The method has the advantages of easiness, conservation of time and low cost; and the prepared feeder-layer cells can be repeatedly used, and cross contamination can be avoided.

The invention discloses a method for efficiently localizing exogenous mammal cytoplasmic protein in a cell nucleus and application thereof. According to the method, a pRK5-Sufu-GFP expression vector is digested by utilizing SalI single enzyme, then NLS is ligated into the vector by utilizing ligase, subsequently nuclear localization vectors pRK5-Sufu-nNLS-GFP (SnNG) with unequal numbers of inserted NLSs are obtained by transformation, and finally cytoplasmic protein is successfully localized in the cell nucleus. In the method disclosed by the invention, multiple NLSs can be inserted at one time, inserted n plasmid vectors (n being 1-5) are obtained simultaneously, and the exogenous cytoplasmic protein can be efficiently localized in the cell nucleus. The transfection of the plasmid into mouse fibroblasts (MEFs) is achieved by one-time construction of the cytoplasmic protein with 1-5 NLSs, so as to study the dose-effect relationship between the localization of the cytoplasmic protein incytoplasm / karyon and the biological activity and function of the cytoplasmic protein in cells.

The invention specifically relates to a chloroquine cholesterol derivative, and a preparation method and application thereof, belonging to the field of the chemical pharmaceutical industry. The chloroquine cholesterol derivative provided by invention has a structure as shown in a formula I which is described in the specification. The invention also provides the preparation method and application of the chloroquine cholesterol derivative. The chloroquine cholesterol derivative provided by the invention can effectively inhibit the proliferation of rat fibroblasts and treat pulmonary fibrosis, exerts anti-inflammatory effect, and offers a novel option for preparation of anti-fibrotic, anti-inflammatory and anti-tumor drugs.

The invention relates to porous cuprous oxide nano crystals, and a preparation method and application thereof. The preparation method comprises following steps: ultrasonically dissolving polyethyleneglycol in a copper acetate monohydrate aqueous solution, adding a sodium hydroxide aqueous solution, adding an ascorbic acid aqueous solution after the reaction is finished, and continuously reactingan obtained mixed solution under the protection of nitrogen after the addition is finished; and then centrifuging the mixed solution, washing with deionized water and ethanol respectively, drying a solid obtained by centrifugation in a vacuum drying oven, and storing in nitrogen. The preparation method of the porous cuprous oxide nano crystals is simple; the raw materials are widely available andcheap. The prepared cuprous oxide nano crystals have an obvious pore channel structures. The prepared porous cuprous oxide nano crystals have a remarkable inhibition effect on tumor cells A549, and have low cytotoxicity on normal cell mouse fibroblasts L929.

The present invention provides a cordyceps sinensis-containing plant composition extraction method, a cordyceps sinensis-containing plant composition is prepared from 70 parts of cordyceps sinensis, 50 parts of neolepisorus ovatus, 60 parts of amaranthus tricolor, 40 parts of Sorbus pohuashanensis and 80 parts of Abelia engleriana (Graebn. ) Rehd as medicinal raw materials by microwave extraction method, the content is greatly improved, the use amount is reduced, and the present invention also provides application of the cordyceps sinensis-containing plant composition in manufacture of rat fibroblast cell line NIH / 3T3 proliferation suppressing drugs.