113 results about "Leukemia inhibitory factor" patented technology

The present invention provides a non-mouse, including human, pluripotential embryonic stem cell which can:(a) be maintained on feeder layers for at least 20 passages; and(b) give rise to embryoid bodies and multiple differentiated cell phenotypes in monolayer culture.The invention further provides a method of making a pluripotential embryonic stem cell comprising culturing germ cells and germ cell progenitors in a composition comprising a growth enhancing amount of basic fibroblast growth factor, leukemia inhibitory factor, membrane associated steel factor, and soluble steel factor to primordial germ cells under cell growth conditions, thereby making a pluripotential embryonic stem cell.Also provided are compositions useful to produce the pluripotent embryonic stem cells and methods of screening associated with the method of making the embryonic stem cell.

Treating or preventing the early stages of degeneration of articular cartilage or subchondral bone in the affected joint of a mammal is accomplished by administering a chondroprotective compound of Formula (I):where A is hydroxy, (C1-C4)alkoxy, amino, hydroxy-amino, mono-(C1-C2)alkylamino, di-(C1-C2)alkylamino; X and Y are independently H or (C1-C2)alkyl; and n is 1 or 2; R6 is halogen, (C1-C3)alkyl, trifluoromethyl, or nitro; R9 is H; (C1-C2)alkyl; phenyl or phenyl-(C1-C2)alkyl, where phenyl is optionally mono-substituted by fluoro or chloro; -C(=O)-R, where R is (C1-C2)alkyl or phenyl, optionally mono-substituted by fluoro or chloro; or -C(=O)-O-R', where R1 is (C1-C2)alkyl.This treatment ameliorates, diminishes, actively treats, reverses or prevents any injury, damage or loss of articular cartilage or subchondral bone subsequent to said early stage of said degeneration. Whether or not a mammal needs such treatment is determined by whether or not it exhibits a statistically significant deviation from normal standard values in synovial fluid or membrane from the affected joint, with respect to at least five of the following substances: increased interleukin-1 beta (IL-1beta); increased tumor necrosis factor alpha (TNFalpha); increased ratio of IL-1beta to IL-1 receptor antagonist protein (IRAP); increased expression of p55 TNF receptors (p55 TNF-R); increased interleukin-6 (IL-6); increased leukemia inhibitory factor (LIF); decreased insulin-like growth factor-1 (IGF-1); decreased transforming growth factor beta (TGFbeta); decreased platelet-derived growth factor (PDGF); decreased basic fibroblast growth factor (b-FGF); increased keratan sulfate; increased stromelysin; increased ratio of stromelysin to tissue inhibitor of metalloproteases (TIMP); increased osteocalcin; increased alkaline phosphatase; increased cAMP responsive to hormone challenge; increased urokinase plasminogen activator (uPA); increased cartilage oligomeric matrix protein; and increased collagenase.

The invention provides a serum-free medium for umbilical cord mesenchymal stem cells and belongs to the technical field of cell culture. The serum-free medium for umbilical cord mesenchymal stem cells comprises a basal culture medium body and added ingredients, wherein the basal culture medium body is a DMEM culture medium; the added ingredients include a basic fibroblast growth factor hFGF, a epidermal growth factor hEGF, insulin hI, a leukaemia inhibitory factor hLIF and astragalus polysaccharide. The umbilical cord mesenchymal stem cells are subjected to subculture and amplification in the medium, and the surfaces of the cultured cells are marked and analyzed. The serum-free medium for umbilical cord mesenchymal stem cells overcomes the defect of exogenous pollution of serum and solves the problem of contradiction between the cell expansion number and cost reduction. The medium is free of serum, thereby preventing influence of animal-derived serum ingredients on cell culture. The medium can be used for studying the differentiation and proliferation adjusting mechanism of the umbilical cord mesenchymal stem cells.

The invention discloses an isolated culture method of human adipose-derived stem cells and a construction method of a stem cell bank. The isolated culture method comprises the following steps of: (1) collecting a human adipose tissue; (2) obtaining and separating the human adipose-derived stem cells; (3) culturing the stem cells; (4) detecting and cryopreserving; and (5) constructing the stem cell bank. According to the invention, the mixed collagenase prepared from D-Hanks balanced salt solution and containing type I and type VI collagenases is employed to digest the adipose tissue so that the tissue is digested more thoroughly, the number of parenchyma cells is obviously reduced and tissue blocks are removed; the MCDB-201 culture solution containing 10-15% of fetal calf serum and 10<3>-10<5>U / Ml LIF (Leukemia Inhibitory Factor) is used for culturing the isolated stem cells so that the cells proliferate quickly and are good in morphology; the differentiation of the stem cells can be effectively inhibited and the characteristics of the primary stem cells are ensured; and in the meantime, the human adipose-derived stem cells can be promoted to grow for a long time, and to keep the features such as self-renewal and multipotential differentiation; and moreover, the obtained stem cells are saved in the bank constructed so that better-quality seed cell source is guaranteed.

The invention discloses an adipose-derived stem cell separation culture method comprising the following steps of: separating and acquiring from adipose tissues; and culturing SVF (Stromal-vascular fraction) cells. The separation culture method is characterized by also comprising the following steps of: removing Lin+cells included in the SVF cells to obtain an Lin-cell mass by using a magnetic-activated cell sorting method; gathering CD271+Sca-1+cells from the obtained Lin-cell mass to obtain the adipose-derived stem cells by using a fluorescent-activated cell sorting method; and culturing theobtained adipose-derived stem cells by using a culture medium containing LIF (Leukemia Inhibitory Factor) and FGF2 (Fibroblast Growth Factor). The invention can efficiently and fast obtain the high-purity adipose-derived stem cells by combining FACS (Fluorescent-Activated Cell Sorting) with MACS (Magnetic-Activated Cell Sorting), wherein the high-purity adipose-derived stem cells have higher clone forming ability, self-renewal ability and multi-directional differentiation potentiality; P16 generation adipose-derived stem cells obtained by the separation culture method still have higher adipose forming ability and bone forming ability; in addition, the invention provides a new method for acquiring a large quantity of seed cells with dryness in vitro to apply to tissue regeneration.

The invention relates to a serum-free adipose tissue-derived mesenchymal stem cell culture medium, which consists of a basic culture medium and added ingredients, wherein the basic culture medium is DMEM-LG, and the added ingredients and the content of each added ingredient are shown as follows: 5 to 20ng / mL of alkaline fiberblast growth factors, 5 to 20ng / mL of epidermal growth factors, 100U / mL of penicillin, 100 micrograms / mL of streptomycin, 50 to 200 micrograms / mL of heparin, 2 to 8mM of L-glutamine, 100 to 300 microM of 2-mercaptoethanol, 500 to 2000U / mL of leukaemia inhibitory factors and 0.5 to 2mM of sodium pyruvate. The serum-free adipose tissue-derived mesenchymal stem cell culture medium does not contain the serum, so that the inter-batch difference and the influence of the serum component on the cell culture can be avoided; the exogenous pollution of the serum and the toxicity of the serum on the cells can be avoided; and the ingredients are clear, so that the research of the psychological regulation mechanism of the cells can be facilitated.

The present invention provides defined serum-free cell culture media useful in culturing fibroblasts, especially articular chondrocytes, that avoid problems inherent in the use of serum-containing media. The defined media comprise platelet-derived growth factor (PDGF), chemically defined lipids, oncostatin M (OSM), interleukin-6 (IL-6), leukemia inhibitory factor (LIF), 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.

The invention provides a novel culture medium useful for the proliferation and maintenance of avian primordial germ cells (PGCs) and encompassing a medium base, leukemia inhibitory factor, basic fibroblast growth factor, stem cell factor and insulin-like growth factor, and an avian serum. The invention also provides a method for the proliferation and maintenance of avian primordial germ cells for extended periods encompassing the steps of isolating a population of PGCs from an avian and culturing the PGCs in a culture medium containing the growth factors and avian serum. The invention further provides methods of producing chimeric avians by isolating and culturing PGCs in a culture medium containing avian serum, transferring the PGCs into a recipient avian embryo, and allowing the recipient avian to develop into a chimeric bird. Another aspect of the invention provides a culture of avian PGCs grown and maintained in the culture medium containing avian serum.

Leukemia Inhibitory Factor (“LIF”), an LIF analogue, an LIF mimetic and a product capable of stimulating the expression of endogenous LIF, and mixtures thereof are useful, for (i) promoting the multiplication, in vitro, of a population of human skin stem cells and / or epidermal progenitors while maintaining them in an undifferentiated state, and / or for (ii) maintaining and / or increasing their capacity to generate a pluristratified epithelium, in particular a pluristratified epidermis and / or all or some of the skin appendages; also a library or a culture of human undifferentiated skin stem cells and / or epidermal progenitors is obtained in the presence of LIF, as are reconstructed epidermides and / or reconstructed skin, and kits for producing libraries of cells or reconstructed epidermides, and stem cells or reconstructed epidermides are prepared for the treatment of individuals exhibiting damaged skin (individuals suffering from third degree burns and individuals suffering from genetic diseases affecting the skin).

The invention relates to a tumor tissue tumor infiltrating lymphocyte (TIL) cell preparation method and a dedicated culture medium. The method comprises the following steps of tumor surrounding tissue obtaining, cell digestion, cell primary culture, cell subculture and cell collection, wherein a primary culture medium is based on a RPMI 1640 culture medium and prepared from the following concentration ingredients of 10% volume of human-derived serum, 20 to 45ng / ml of basic fibroblast growth factor (bFGF), 1 to 5mg / ml of riboflavin, 70 to 90ng / ml of cortisol, 10 to 25mg / ml of sodium dihydrogen phosphate monohydrate, 47 to 62ng / ml of recombinant human leukaemia inhibitory factor (LIF) and 500 to 800U / ml of IL-2; a subculture medium is based on the RPMI 1640 culture medium and prepared from the following concentration ingredients of 10% volume of the human-derived serum, 20 to 40mmol / L of HEPES, 1000 to 2000U / ml of the IL-2, 0.03 to 0.07mmol / L of beta-mercaptoethanol and 5 to 15ng / ml of sodium phosphate. According to the preparation method, an existing culture medium is improved, different culture mediums are utilized to culture the TIL cells in pertinence, TIL cell expansion capacity is improved, meanwhile a culture period is reduced, a culture complexity degree is reduced, a use amount of the IL-2 is reduced, and toxic reaction is reduced.

The invention relates to a method for production of neurons from cells of a cell line which may be differentiated to produce neurons in particular, whereby said cells are cultivated in spheres, preferably by exposing the same to growth factors, such as, for example, EGF (epidermal growth factor) and / or bFGF (basic fibroblast growth factor) or LIF (Leukemia Inhibitory Factor), in a given growth medium, the differentiation in said spheres is induced on forcing the same to adhere to a substrate, after removal of the growth factors EGF and / or bFGF or LIF, and cultivating the same in the growth medium for an appropriate duration. Said method is characterised in that cells of the human embryonic teratocarcinoma NT2 are used.

The invention provides methods for obtaining neural stem cells from a mammalian embryonic or inducible pluripotent stem cell population comprising culturing mammalian embryonic or inducible pluripotent stem cells in a cell culture medium having a leukemia inhibitory factor (LIF), an inhibitor of glycogen synthase kinase 3 (GSK3), and an inhibitor of transforming growth factor β (TGF-β) under suitable conditions and obtaining isolated neural stem cells therefrom.

Human pre-formed xenoantibodies play an important role in the hyperacute rejection response in human xenotransplantation. Disclosed are materials and methods for removing or neutralizing such antibodies. Also disclosed are materials and methods for reducing or eliminating the epitopes in the donor organs that are recognized by such antibodies. Such epitopes are formed as the result of activity by the enzyme α-1,3 galactosyltransferase. The porcine gene encoding α-1,3 galactosyltransferase is disclosed, as are materials and methods for inactivating (“knocking out”) the α-1,3 galactosyltransferase gene in mammalian cells and embryos. Included are nucleic acid constructs useful for inactivating the α-1,3 galactosyltransferase gene in a target cell. Also disclosed is a novel leukemia inhibitory factor (T-LIF) that is useful for maintenance of embryonic stem cells and primordial germ cells in culture.

Leukemia inhibitory factor (“LIF”), an LIF analogue, an LIF mimetic, a product capable of stimulating the expression of endogenous LIF and mixtures thereof are useful for maintaining and / or stimulating the regenerative power of a population of human undifferentiated skin stem cells and / or epidermal progenitors, and are also useful for treating dry and / or chapped and / or aged skin and / or hair loss, as well as for treating skin burns and promoting skin regeneration and / or cicatrization.

The invention provides methods for obtaining neural stem cells from a mammalian embryonic or inducible pluripotent stem cell population comprising culturing mammalian embryonic or inducible pluripotent stem cells in a cell culture medium having a leukemia inhibitory factor (LIF), an inhibitor of glycogen synthase kinase 3 (GSK3), and an inhibitor of transforming growth factor β (TGF-β) under suitable conditions and obtaining isolated neural stem cells therefrom.

There are provided compositions and methods for treatment of neurodegeneative diseases and CNS injury. The compositions a pharmaceutically acceptable carrier solution; and a plurality of biodegradable nanoparticles, wherein the nanoparticles comprise a targeting moiety that is able to bind selectively to the surface of a neural stem cell and wherein the nanoparticles further comprise factors such as leukaemia inhibitory factor (LIF); XAV939 and / or one or more of : brain-derived neurotrophic factor (BDNF) or an agonist thereof; epidermal growth factor (EGF) or an agonist thereof; glial cell-derived neurotrophic factor (GDNF) or an agonist thereof; retinoic acid and derivatives thereof; ciliary neurotrophic factor (CTNF) or an agonist thereof; and Wnt5A. The biodegradable nanoparticles may deliver via controlled time release.

This document provides methods and materials for treating cardiovascular tissue. For example, stem cells, compositions containing stem cells, methods for obtaining stem cells, compositions for generating stem cells expressing particular markers (e.g., compositions comprising TGF-β1, BMP-2, FGF-4, and leukemia inhibitory factor), and methods for repairing cardiovascular tissue are provided.

The present invention relates to a pharmaceutical composition comprising of preparations of human embryonic stem (hES) cells and their derivatives and methods for their transplantation into the human body, wherein transplantation results in the clinical reversal of symptoms, cure, stabilization or arrest of degeneration of a wide variety of presently incurable and terminal medical conditions, diseases and disorders. The invention further relates to novel processes of preparing novel stem cell lines which are free of animal products, feeder cells, growth factors, leukaemia inhibitory factor, supplementary mineral combinations, amino acid supplements, vitamin supplements, fibroblast growth factor, membrane associated steel factor, soluble steel factor and conditioned media. This invention further relates to the isolation, culture, maintenance, expansion, differentiation, storage, and preservation of such stem cells.

The present invention provides a method of producing pluripotent stem cells, which comprises culturing testis cells using a medium containing glial cell derived neurotrophic factor (GDNF) or an equivalent thereto to obtain pluripotent stem cells. The medium can further contain leukemia inhibitory factor (LIF), epidermal growth factor (EGF), basic fibroblast growth factor (bFGF) and the like. Using the production method of the present invention, it is possible to produce pluripotent stem cells, which have conventionally been only obtainable from fertilized eggs, embryos and the like, from a postnatal individual. Using the pluripotent stem cells, it is possible to construct diverse tissues having histocompatibility for autotransplantation, and the pluripotent stem cells are useful in medical fields such as regeneration medicine and gene therapy. Also, the pluripotent stem cells are useful in the field of biotechnology because they can be used to prepare transgenic animals, knockout animals and the like.

The invention provides a composition for embryonic stem cell culture. The composition can guarantee that embryonic stem cells are in an undifferentiated state for a long term and have the ability of infinite proliferation. The composition comprises leukaemia inhibitory factors (LIF), basic fibroblast growth factor (bFGF), Rho kinase inhibitor Y27632 and bone morphogenetic protein inhibitor Noggin. The invention further provides cell culture liquid containing the composition and application of the composition.

This invention discloses a method for producing liver cells by inducing haemopoietic stem cells in vitro, comprising: inoculating the haemopoietic stem cells or CD34+ cells into the culture media containing embryonic bovine serum, human stem cell factor, human interleukins 3(IL-3), interleukins 6(IL-6), human liver cell growth factor, human epidermal growth factor(EGF), human acdic fibroblast growth factor, human basic fibroblast growth factor(bFGF), human leukemia inhibitory factor(LIF) and oncostatin M(OSM), and inducing the producing of liver cells in vitro. By applying the method of this invention to induce liver cells from the haemopoietic stem cells, it can acquire a high proportion of liver cells in the culture with good effects.

The invention provides a culture medium of umbilical cord mesenchymal stem cells. The culture medium comprises a DMEM (Dulbecco's Modified Eagle Medium) / F12 culture medium, an adding component added into the DMEM / F12 culture medium and ivy extract, wherein the adding component is prepared from the following components: human serum albumin, transferrin, a vascular endothelial growth factor, a basicfibroblast growth factor, arginine, a leukemia inhibitory factor, a stem cell factor, beta cytokine, parathyroid hormone, a platelet-derived growth factor, a tumor necrosis factor alpha, interleukin,erythropoietin, thrombopoietin, alpha-D-glucose, vitamin C, resveratrol, NaHCO3 and ganglioside; the ivy extract is liquid and is extracted from leaves of ivy through a conventional method; the finalconcentration is 20 to 50mg / L after the ivy extract is added. By adopting the culture medium, the problems in the prior art are solved, and the cells can keep good state and proliferation speed.

The present invention provides a method of growing spermatogonial stem cells of mammals and the like in vitro, which is characterized in that glial cell-derived neurotrophic factor (GDNF) or an equivalent thereto, and leukemia inhibitory factor (LIF) are contained in a medium (culture broth) for culturing spermatogonial stem cells. According to the method of the present invention, spermatogonial stem cells can be grown in vitro to the extent that enables use thereof for developmental engineering.

The invention relates to antibodies directed against human Leukemia Inhibitory Factor (LIF) and to a hybridoma cell line producing said antibodies. The invention also relates to a method for blocking / inhibiting the proliferation of stem cells, and to an in vitro method for the diagnosis of diseases associated with unwanted cell proliferation in a subject or for determining the predisposition of a subject to suffer from said disease associated with unwanted cell proliferation, or for prognosis of average life expectancy of a subject suffering from said disease. The therapeutic potential of said antibodies is based on observing that the inhibition of LIF can be used in therapeutic compositions for the treatment of diseases associated with unwanted proliferation.

The present invention relates to a method of producing radial glial cells from neural stem cells, particularly by contacting neural stem cells with epidermal growth factor (EGF), fibroblast growth factor 2 (FGF-2) and / or TGFα. Leukemia inhibitory factor (LIF) and ciliary neurotrophic factor (CNTF) can optionally be added to enhance the effect of EGF, FGF-1 or TGFα. Also provided are methods of producing radial glial cells from ependymal cells, as well as methods of proliferating ependymal cells.

The present invention provides a novel oocyte maturation medium or / and embryo culture medium with a chemically defined supplement to produce matured oocytes at high efficiency. The inventive medium or supplement comprises three growth factors, namely, fibroblast growth factor 2 (FGF2), leukemia inhibitory factor (LIF), and insulin-like growth factor 1 (IGF-1) in a synergistic combination. Methods for oocyte and embryo culture are also provided.

A composition for modulating the immune response in a mammal comprising a pharmaceutically acceptable carrier solution and a plurality of biodegradable nanoparticles, wherein the nanoparticles comprise a targeting moiety that is able to bind selectively to the surface of a T lymphocyte cell and / or of a vascular endothelial cell and wherein the nanoparticles further comprise leukaemia inhibitory factor (LIF). Nanoparticle-mediated targeted delivery of LIF can be used a means to guide tolerogenesis in a patient and has immediate clinical application for recipients of organ grafts and also for patients suffering from autoimmune disease.

The invention provides a mass culture method of umbilical cord mesenchymal stem cells. The method comprises the following steps: obtaining umbilical cord Wharton's jelly; carrying out P0 generation culture on the umbilical cord Wharton's jelly until the cell fusion degree is 40-50%, and carrying out P1 subculture until the cell fusion degree is 85-95%; carrying out inoculating culture on P1 generation cells in a plate medium, and separating suspension cells non-adhered to the plate medium; and carrying out P2 subculture on the suspension cells in a medium containing a leukemia inhibition factor and a fibroblast-like growth factor. The above tissue adherent culture method allows adherence screening to be carried out through using a tissue culture plate after direct culture of P0 cells with low polytropy in early stage into P1 generation in order to maintain the uniformity of the cell differentiation degree; and the leukemia inhibition factor and the fibroblast-like growth factor are used to increase the adherence performance of the cells in the P2 generation culture process and inhibit differentiation, so the obtained umbilical cord mesenchymal stem cells have the advantages of low differentiation degree, trending to uniformity of the differentiation degree, strong adherence capability and collection facilitation.

The present invention relates to methods, compositions, and kits for maintenance of the undifferentiated state and / or pluripotency in stem cells, including embryonic stem (ES) cells. It also relates to stem cells that have been maintained in an undifferentiated state. The present invention provides these aspects using culture medium enriched with a TGFbeta family member protein, an FGF family member protein, and / or nicotinamide, without using fibroblast feeder layers, conditioned media, or leukemia inhibitory factor.