456 results about "Serum free media" patented technology

Disclosed and claimed is a new insect cell line, Sf900+, ATCC CRL-12579. The insect cell line was established from Lepidoptera, Noctuidae, Spodoptera frugiperda Sf-9 (ATCC CRL-1711) through multiple rounds of limiting dilution and selection in a serum-free insect medium supplemented with added human insulin. The insect cell line is useful in BEVS or as an adjuvant and has many characteristics and advantages. Also disclosed and claimed are recombinant proteins from recombinant baculovirus expression in insect cells such as Sf900+ cells, for instance, HA, NA, EPO, CD4, CEA, and thrombospondin.

The present invention relates to a process for producing a desired polypeptide using rat cells. Specifically, the present invention relates to a process for producing the polypeptide which comprises culturing rat cells such as YB2 / 3HL.P2.G11.16Ag.20 (hereinafter referred to as YB2 / 0), preferably rat cells to which a recombinant DNA comprising DNA encoding a desired polypeptide such as an immunologically functional molecule is introduced, in a medium which does not contain serum (hereinafter referred to as a serum-free medium). Among the desired polypeptides obtained by the process of the present invention, an antibody obtained by using a transformant of YB2 / 0 has a high antibody-dependent cell-mediated cytotoxic activity (hereinafter sometimes referred to as ADCC activity) and is useful as a pharmaceutical agent.

The present invention provides defined serum-free cell culture media useful in culturing fibroblasts, especially articular chondrocytes, that avoids problems inherent in the use of serum-containing media. The defined media comprise platelet-derived growth factor (PDGF), and chemically defined lipids, or combinations of these compounds. In another aspect, the present invention also provides tissue culture methods that comprise incubating chondrocytes in the defined serum free media. The methods enhance attachment and proliferative expansion of chondrocytes seeded at low density while maintaining their redifferentiation potential.

Disclosed are compositions and methods for increasing the longevity of a cell culture and permitting the increased production of proteins, preferably recombinant proteins, such as antibodies, peptides, enzymes, growth factors, interleukins, interferons, hormones, and vaccines. Cells transfected with an apoptosis-inhibiting gene or vector, such as a triple mutant Bcl-2 gene, can survive longer in culture, resulting in extension of the state and yield of protein biosynthesis. Such transfected cells exhibit maximal cell densities that equal or exceed the maximal density achieved by the parent cell lines. Transfected cells can also be pre-adapted for growth in serum-free medium, greatly decreasing the time required to obtain protein production in serum-free medium. In certain methods, the pre-adapted cells can be used for protein production following transfection under serum-free conditions. In preferred embodiments, the cells of use are SpESF or SpESF-X cells.

The present invention relates to methods for modulating the glycosylation profile of recombinantly-expressed proteins. In particular, the present invention relates to methods of controlling the galactosylation profile of recombinantly-expressed proteins by supplementing production medium, e.g., a hydrolysate-based or a chemically defined medium, with manganese and / or D-galactose.

Serum-free media for growth and proliferation of chondrocytes and mesenchymal stem cells in culture are provided. A serum-free medium for growth of chondrocytes includes a serum-free composition comprising FGF-2, linoleic acid, ascorbic acid, B-mercaptoethanol, transferrin and dexamethasone. Further, the composition comprises EGF, PDGFbb, insulin and albumin. A method for growing chondrocytes in a serum free medium comprising the compostion is also provided. Also provided for mesenchymal stem cell growth, is a serum-free medium which includes a composition comprising FGF-2, LIF, SCF, pantotenate, biotin and selenium and method, therefore.

Synthetic surfaces capable of supporting culture of eukaryotic cells including stem cells and undifferentiated human embryonic stem cells in a chemically defined medium include a swellable (meth)acrylate layer and a polypeptide conjugated to the swellable (meth)acrylate layer. The swellable (meth)acrylate layer may be formed by polymerizing monomers in a composition that includes a carboxyl group-containing (meth)acrylate monomer, a cross-linking (di- or higher-functional) (meth)acrylate monomer, and a hydrophilic monomer capable of polymerizing with the carboxyl group-containing (meth)acrylate monomer and the cross-linking (meth)acrylate monomer. The swellable (meth)acrylate layer has an equilibrium water content in water of between about 5% and about 70%. The conjugated peptide may include an RGD amino acid sequence.

Synthetic surfaces capable of supporting culture of undifferentiated human embryonic stem cells in a chemically defined medium include a swellable (meth)acrylate layer and a peptide conjugated to the swellable (meth)acrylate layer. The swellable (meth)acrylate layer may be formed by polymerizing monomers in a composition that includes hydroxyethyl methacrylate, 2-carboxyehylacrylate, and tetra(ethylene glycol) dimethacrylate. The conjugated peptide may include an amino acid sequence of XaanProGlnValThrArgGlyAspValPheThrMetPro, where n is an integer from 0 to 3 and where Xaa is any amino acid. Further, disclosed herein is a swellable (meth)acrylate synthetic surface which can be sterilized by gamma irradiation.

Methods and devices for culturing human pluripotent stem cells to produce cells of the pancreatic lineage are disclosed. The methods include steps of culturing the stem cells under conditions that induce the expression of mesendoderm / primitive streak and definitive endoderm markers in a chemically defined medium including an effective amount of i) fibroblast growth factor, ii) Activin A, and iii) bone morphogenetic protein. The methods further include the steps of culturing cells under conditions favoring the formation of at least one of intact embryoid bodies and pancreatic progenitor PDX1+ Ins− cells.

The invention is a cell culture medium that can include reduced or no serum and that enhances the performance of serum-free media for cell culture. The medium supports the growth of cells for both small scale and large scale propagation of cells. The invention also includes a method of cultivating cells using the cell culture medium of the present invention.

The present invention belongs to the field of biotechnology, and discloses a serum-free culture medium with specific chemical compositions for in vitro culture and amplification of bone marrow mesenchymal stem cells. By adding insulin, transferrin, ethanolamine, sodium selenite, growth factors, adherent factors, hormone, putrescine, inorganic salt, vitamin, albumin and antioxidant into a basic culture medium, the bone marrow mesenchymal stem cells can attach to the culture medium under a serum free condition, so the in vitro culture and amplification are realized, the potential of multi-directional differentiation is maintained, and the amplified cells can be induced to be osteoblast and lipocyte in vitro. The serum-free culture medium has the advantages that the clinic level cell products for human produced by the serum free culture medium can effectively avoid the potential risk of producing cell products by serum culture medium. The drawing appended is a photo of the confluence of the bone marrow mesenchymal stem cells cultured by the serum-free culture medium.

Disclosed is a cervico-vaginal tissue equivalent comprised of vaginal epithelial cells and immune cells, cultured at the air-liquid interface. The tissue equivalent is capable of being infected with a sexually transmitted pathogen such as a virus (e.g., HIV), a bacteria, a helminthic parasite, or a fungus. The tissue equivalent is also capable of undergoing an allergic-type reaction or an irritant-type reaction. The tissue equivalent is characterized as having nucleated basal layer cells and nucleated suprabasal layer cells, and further as having cell layers external to the suprabasal layer progressively increasing in glycogen content and progressively decreasing in nuclei content. Immune cells of the tissue equivalent are primarily located in the basal and suprabasal layers. Also disclosed are methods for producing the tissue equivalent. The methods involve providing vaginal epithelial cells and immune cells, seeding the cells onto a porous support, and co culturing the seeded cells at the air-liquid interface under conditions appropriate for differentiation. One such method disclosed is for generation of the tissue equivalent in serum free medium. Specific cells from which the tissue equivalent is generated, and also specific preferred components of the medium in which the tissue equivalent is generated are provided. Also disclosed is a cervico-vaginal tissue equivalent produced by the methods disclosed herein.

The present invention provides an attenuated virus, which is derived from Modified Vaccinia Ankara virus and characterized by the loss of its capability to reproductively replicate in human cell lines. It further describes recombinant viruses derived from this virus and the use of the virus, or its recombinants, as a medicament or vaccine. A method is provided for inducing an immune response in individuals who may be immune-compromised, receiving antiviral therapy, or have a pre-existing immunity to the vaccine virus. In addition, a method is provided for the administration of a therapeutically effective amount of the virus, or its recombinants, in a vaccinia virus prime / vaccinia virus boost innoculation regimen. The present invention relates to a method of virus amplification in primary cells which are cultivated in a serum free medium. Viruses produced by this method are advantageously free of any infectious agents comprised in animal sera.

The invention relates to the culture medium research and development technical field of modern biological technology and provides a serum-free medium for MDCK cell large-scale adherent culture and single-cell suspension culture, which comprises 21 amino acids, 6 vitamins, 8 salts, 8 lipids, 4 trace elements, 2 buffers, 1 protein hydrolysate, 1 acid-base indicator and 6 other additives. The serum-free medium can be prepared by the conventional preparation method, and an application method thereof is the conventional method. The serum-free medium has the beneficial effects that: the serum-free medium does not contain serum, has clear components, is beneficial for separating and purifying the product and improves the product quality; the serum-free medium supports long-term subculture of MDCK cells and does not require long-term and complex adaptation process; and the serum-free medium can well support the adherent growth and single-cell suspension growth of the MDCK cells, has clear components and easy preparation and utilization, and is suitable for mass production of biological products.

The present invention provides a clinically applicable method of inducing differentiation of embryonic stem cells, particularly a method of inducing differentiation of embryonic stem cells into forebrain neurons. More specifically, the present invention provides a method of inducing differentiation of embryonic stem cells, comprising culturing the embryonic stem cells as a floating aggregate in a serum-free medium, particularly a method of inducing differentiation of the embryonic stem cells into nervous system cells such as forebrain neurons and cerebellar neurons and sensory organ cells; a floating aggregate of embryonic stem cells obtained by culturing the embryonic stem cells as a floating aggregate in a serum-free medium; and cells derived from a floating aggregate of embryonic stem cells, particularly nervous system cells such as forebrain neurons and cerebellar neuron, sensory organ cells such as retinal precursor cells, and the like.

A method of establishing a clonal embryonic stem cell line capable of sustaining a phenotype of normal embryonic stem cells following at least eight months of in vitro culture is disclosed. The method is effected by culturing an individual embryonic stem cell for at least eight months in a serum-free medium, thereby establishing the clonal embryonic stem cell line capable of sustaining said phenotype of normal embryonic stem cells following at least eight months of in vitro culture.

This invention relates to human adipose tissue-derived multipotent adult stem cells. More particularly, the invention relates to human adipose tissue-derived multipotent stem cells, which can be maintained in an undifferentiated state for a long period of time by forming spheres and have high proliferation rates, as well as methods for isolating and maintaining the adult stem cells, and methods for differentiating the multipotent adult stem cells into nerve cells, fat cells, cartilage cells, osteogenic cells and insulin-releasing pancreatic beta-cells. Also, the invention relates to cellular therapeutic agents for treating osteoarthritis, osteoporosis and diabetes and for forming breast tissue, which contain the differentiated cells or the adult stem cells. Although the multipotent stem cells are adult stem cells, they have the ability to differentiate into osteogenic cells, nerve cells, astrocytes, fat cells, chrondrogenic cells or insulin-releasing pancreatic beta-cells, and so are effective in treating osteoporosis, osteoarthritis, nerve disease, diabetes, etc. Also, the stem cells form spheres in a serum-free medium containing CORM-2, and thus can be maintained in an undifferentiated state for a long period of time. Also, the stem cells have very high proliferation rates. Accordingly, the stem cells are useful as cellular therapeutic agents.

The present invention provides a method of efficiently expanding non-adhesive adult stem cells in vitro. More specifically, the present invention provides a method of expanding non-adhesive adult stem cells, comprising culturing non-adhesive adult stem cells in the co-presence of adhesive feeder cells and non-adhesive non-stem cells in serum-free medium, wherein the ratio of non-adhesive adult stem cell count to total cell count in the medium is kept low; non-adhesive adult stem cells obtained by the expansion method and differentiated cells thereof, and a composition comprising them; and a serum-free medium comprising a specified factor, and a kit for its preparation and the like.

The invention relates to the preparation and application method of a medium, which is specifically related to a medium which is used for Vero cell and a culture method of the Vero cell. The invention belongs to the biological product area. The culture method of the invention is that the adhering Vero cells are suspend and cultured in SFM serum-free medium for the domestication and adaptation; the growing pattern of the Vero cell changes from adherence growth to suspension growth; the Vero cells are further amplified in Vero amplification CHEMICALLY DEFINED MEDIUM. The method of the invention has good effects; the growing pattern of the Vero cell is transformed from adherence growth to suspension growth, which changes the growth pattern of the cell and greatly increases the yield of the cell.

This invention relates to human adipose tissue-derived multipotent adult stem cells. More particularly, the invention relates to human adipose tissue-derived multipotent stem cells, which can be maintained in an undifferentiated state for a long period of time by forming spheres and have high proliferation rates, as well as methods for isolating and maintaining the adult stem cells, and methods for differentiating the multipotent adult stem cells into nerve cells, fat cells, cartilage cells, osteogenic cells and insulin-releasing pancreatic beta-cells. Also, the invention relates to cellular therapeutic agents for treating osteoarthritis, osteoporosis and diabetes and for forming breast tissue, which contain the differentiated cells or the adult stem cells. Although the multipotent stem cells are adult stem cells, they have the ability to differentiate into osteogenic cells, nerve cells, astrocytes, fat cells, chrondrogenic cells or insulin-releasing pancreatic beta-cells, and so are effective in treating osteoporosis, osteoarthritis, nerve disease, diabetes, etc. Also, the stem cells form spheres in a serum-free medium containing CORM-2, and thus can be maintained in an undifferentiated state for a long period of time. Also, the stem cells have very high proliferation rates. Accordingly, the stem cells are useful as cellular therapeutic agents.

Adipose tissue-derived stromal cells induced to form multicellular aggregates can be formed reliably and consistently, they can be maintained for prolonged periods in adherent or suspension culture, and they are able to survive, grow, and / or differentiate in serum-free media conditions. The present invention provides compositions and methods for the use of such aggregates in tissue repair. This culture platform provides a controlled and defined system in which to study and standardize adult stem cell biology, and begets an instinctive “modular” approach to the predictable replacement and regeneration of adipose tissue.

The invention relates to a method for in-vitro amplification of NK cells, and in particular relates to a method for massive in-vitro amplification of NK cells, wherein the method comprises the following steps of: a, inoculating a peripheral blood mononuclear cell in a CD3McAb and CD226McAb pre-coated culture bottle for coculture; b, adding 1L-2 and 1L-18, coculturing for 72hours to stimulate amplification of NK cells; c, transferring the NK cells, K562 cells after lethal treatment and a serum-free medium containing 1L-2 and 1L-18 in a cell culture bag for coculture; and d, collecting the NK cells. According to the method for in-vitro amplification of the NK cells, two antibodies CD3McAb and CD226McAb are simultaneously coated, so the cell factor synthesis and ADCC effect are promoted, and killing toxicity of the NK cells is remarkably improved; the activation and amplification on the NK cells are achieved just by the 1L-2 and 1L-18 cell factors, so the amplification multiple and cell toxicity of the NK cells are guaranteed, and the cost of cell culture is reduced.

The invention discloses a method for simultaneous and efficient amplification of CD<3+>CD<56+>CIK cells and CD<3->CD<56+>NK cells. The method comprises the steps as follows: the concentration of separated PBMC (peripheral blood mononuclear cells) is adjusted by a serum-free medium containing autologous plasma, an Anti-CD16 antibody, IL-2 and IL-15 are added, and then the mixture is transferred into a T175 culture flask for culture; an Anti-CD3 antibody and an Anti-CD137 antibody are added; a serum-free medium containing the autologous plasma, IL-2 and IL-15 is supplemented every two days according to the cell growth condition; the cell concentration is controlled to be about 1.5*10<6> / ml; and after culture is performed for 14-21 days, large quantities of high-purity CD<3+>CD<56+>CIK cells and CD<3->CD<56+>NK cells can be obtained simultaneously, and the total cell quantity can reach an effective value of the cell quantity required for adoptive cellular immunotherapy clinically for tumor. The method for simultaneous and efficient amplification of the CD<3+>CD<56+>CIK cells and the CD<3->CD<56+>NK cells is simple, convenient, effective and high in cell killing activity.

The present invention describes a serum-free medium that promotes the growth and differentiation of erythroid cells, cells that are highly transducible by a non-viral method and genes which increase the growth of erythroid cells and decrease their dependency on Epo. This invention can be used in the expansion of hematopoietic stem cells to produce cultures of erythroid cells as a source of erythroid-specific proteins such as hemoglobin. Hematopoietic stem cells are cultured ex vivo in a serum-free culture medium with the addition of IL-3, SCF and EPO. Cells transfected with the gene described in the present invention can be cultured in the serum-free culture medium with decreased dependency on Epo and other cytokines, thereby reducing the cost of the production of hemoglobin.

The invention discloses an efficient multiplication CTL preparation method killing tumors in a targeted mode. The CTL preparation method comprises the following steps: (a) removing CD4+CD25+Treg cells through immunomagnetic bead negative sorting; (b) arranging mixed cells in a serum-free medium for cultivation, and obtaining suspension cells and adherent cells; (c) adding GM-SCF and IL-4 in the adherent cells, culturing the cells for five days; in the sixth day, adding a tumour cell holoantigen, and in the seventh day, adding TNF-alpha and IL-27; (d) transferring the suspension cells to a culture flask wrapped by a CD3 monoclonal antibody and recombinant human fibronectin, adding IFN-gamma, in the second day, adding IL-2, IL-12 and the IL-27, and culturing the mixture till the eighth day to obtain CIK cells; (e) mixing the CIK cells and mature DC cells, and adding the IL-12, IL-7 and an anti-CD 28 monoclonal antibody for cultivation; in the third day, adding an anti-CTLA-4 monoclonal antibody, and then culturing the mixture for four days. According to the efficient multiplication CTL preparation method killing tumors in the targeted mode, efficiency of in-vitro CTL cell proliferation is improved, activity of killing the tumor cells in the targeted mode is improved, transformation of peripheral blood mononuclear cells to the CD4+CD25+Treg cells is inhibited.

Chemically defined media compositions for the culture of eukaryotic cells are disclosed. The compositions are useful for eukaryotic cell culture in perfusion bioreactors and other vessels.