Abstract: The present invention relates to a method for in vitro maturation of oocytes comprising the steps of: (a) culturing one or more GV oocytes in a culture medium, the culture medium comprising a nuclear maturation inhibiting substance and comprising one or more gonadotropins and/or one or more growth factors, the culturing taking place for a time period sufficient for cytoplasmatic maturation to occur; (b) washing the GV oocytes of step (a) to remove the nuclear maturation inhibiting substance; (c) culturing the washed oocytes of step (b) in a culture medium comprising one or more gonadotropins and/or one or more growth factors and/or MAS for a time period sufficient for nuclear maturation. The invention also relates to an oocyte culture medium comprising a nuclear maturation inhibiting substance and comprising one or more gonadotropins and/or one or more growth factors.

Abstract: The invention provides a method for using collagenase to dissociate neural stem cells in neural stem cell cultures. The collagenase treatment results in an increased cell viability and an increased number of proliferated neural stem cells over time.

Abstract: This invention relates to a hematopoietic stem cell proliferating agent comprising IGF-I, a hematopoietic stem cell proliferating agent comprising IGF-I and at least one protein selected from among SCF, M-CSF, and G-CSF, and a method of growing hematopoietic stem cells which comprises culturing hematopoietic stem cells in a medium containing IGF-I and at least one protein selected from the group consisting of SCF and M-CSF. The hematopoietic stem cell proliferating agent of the invention causes hematopoietic stem cells to proliferate in the undifferentiated state whether in vivo or in vitro and can, therefore, be used for amelioration of the cytopenia induced by radiotherapy or chemotherapy using anticancer drugs, prevention of infectious diseases associated with lymphopenia, or in vitro culture for multiplication of hematopoietic stem cells and extrasomatic culture of recombinant stem cells in gene therapy.

Abstract: We describe a new two-step culture method for mass production in vitro of erythroid cells from either CD34+ (105 cells/mL) or light-density (106 cells/mL) cells purified from the blood of normal donors and thalassemic patients. The method includes (i) culture of the cells in the presence of dexamethasone and estradiol (10?6 M each) and (ii) the growth factors SCF (50 ng/mL), IL-3 (1 ng/mL), and EPO (1 U/mL). In their proliferative phase, these cultures generated about 1-2×107 erythroblasts for each milliliter of blood collected from normal donors or thalassemic patients. They were composed mostly (90%) of CD45low/glycophorin (GPA)neg/CD71low cells at day 7, 50-60% of which became CD45neg/GPA+/CD71high by days 15-20. However, when cells from days 7 to 12 of the proliferative phase were transferred in differentiation medium containing EPO and insulin, they progressed to mature erythroblasts (>90% benzidinepos and CD45neg/GPA+/CD71medium) in 4 days.

Abstract: The present invention provides methods and compositions for providing graft recipients with epidermal melanocytes. Specifically, the methods and compositions of the invention provide for populations of epidermal melanocytes that may be isolated from a patient and cultured in vitro to generate a proliferating population of epidermal melanocytes that exhibit growth, migration and melanin production. The invention is based on the discovery of a culture medium composed of natural components, for culturing epidermal melanocytes that exhibit increased proliferative capacity, migratory behaviors and melanin production. The invention provides novel in vitro methods for culturing epidermal melanocytes, including those isolated from the skin of a healthy subject, to generate a proliferating population of epidermal melanocytes. The methods and compositions of the invention may be used for transplantation to treat patients having hypopigmentation or depigmentation skin disorders.

Abstract: The invention features methods of promoting hair growth in a subject. The methods include inducing or mimicking the effects of Wnt promoted signal transduction, e.g., by increasing the level of Wnt protein or administering an agent which mimics an effect of Wnt promoted signal transduction, e.g., by administering lithium chloride. Methods of inhibiting hair growth are also provided.

Abstract: TALL-104 cells, and other cytotoxic T cell lines, may be modified to increase the cytotoxicity thereof, to enhance growth properties, and/or to provide a preferred phenotype, e.g., expression of cell surface antigens, function, e.g., change in cytokine production profile, by culturing the cells in an effective amount of IL-15, optionally followed by gamma irradiation to halt proliferation.

Abstract: The present invention provides a serum-free supplement which supports the growth of hematopoietic cells in culture. Also provided are a medium comprising a basal medium supplemented with the serum-free supplement of the present invention. The present invention also provides methods for culturing and for differentiating hematopoietic cells.

Abstract: Surgically implantable tissue engineered intervertebral disc tissues that effectively replicate the physicochemical properties of the corresponding in vivo tissues are provided. Methods for producing such tissues can involve culturing the intervertebral disc cells to produce cells surrounded by an extracellular matrix and culturing the cells and matrix on a semipermeable membrane to form a cohesive tissue.

Abstract: The present invention relates to a substantially pure population of viable pancreatic progenitor cells, and methods for isolating such cells. The present invention further concerns certain therapeutic uses for such progenitor cells, and their progeny.

Abstract: The present invention relates to a medium for promoting the survival of islet cells, which comprises one or more growth factors in combination with FK506 in amounts having an anti-apoptotic effect on islet cells in a physiologically acceptable culture medium.

Abstract: A method for culturing Langerhans islets to obtain an amount sufficient for transplant and autotransplant is disclosed. The islets are cultured in a culture serum (rat/human) medium which is supplemented with radical scavengers, growth factors, a matrix material, nerve growth factor, cell migrating/scattering factors and anti-integrin &bgr;1 antibody at proper the time during the culturing process The medium is supplemented with radical scavengers and growth factors for the first time and then further supplemented with matrix material, radical scavengers, nerve growth factor and the growth factors around 12-24 hours after culturing. Thereafter, the medium is supplemented with growth factors, cell migrating/scattering factors and anti-integrin &bgr;1 antibody at 4-5 days into the culturing process.

Abstract: The present invention relates to a method for producing dendritic cells from human hematopoietic progenitor cells by obtaining a cell sample that includes human progenitor cells and culturing the cell sample under plasma-free and serum-free conditions in the presence of a combination of cytokines to produce dendritic cells, where the combination of cytokines include TGF-&bgr;1.

Abstract: This invention relates to a hematopoietic stem cell proliferating agent comprising IGF-I, a hematopoietic stem cell proliferating agent comprising IGF-I and at least one protein selected from among SCF, M-CSF, and G-CSF, and a method of growing hematopoietic stem cells which comprises culturing hematopoietic stem cells in a medium containing IGF-I and at least one protein selected from the group consisting of SCF and M-CSF. The hematopoietic stem cell proliferating agent of the invention causes hematopoietic stem cells to proliferate in the undifferentiated state whether in vivo or in vitro and can, therefore, be used for amelioration of the cytopenia induced by radiotherapy or chemotherapy using anticancer drugs, prevention of infectious diseases associated with lymphopenia, or in vitro culture for multiplication of hematopoietic stem cells and extrasomatic culture of recombinant stem cells in gene therapy.

Abstract: A method for preparing a preparation of mammalian pancreatic endocrine cells comprising the steps: dissociating intact pancreatic tissue into a cell suspension comprising single cells and cell aggregates; enriching said cell suspension with regard to the content in endocrine cells by separating single cells and cellular aggregates with size <100 &mgr;m; and removing contaminating non-endocrine cells by density centrifugation.

Abstract: A chemically defined mammalian cell culture medium is provided that supports maintenance and long term clonal growth of mammalian hepatocytes and other cells.

Abstract: A chemically defined mammalian cell culture medium is provided that supports maintenance and long term clonal growth of mammalian hepatocytes and other cells.

Abstract: A method of generating neural crest stem cells involves inducing neuroepithelial stem cells to differentiate in vitro into neural crest stem cells. Differentiation can be induced by replating the cells on laminin, withdrawing mitogens, or adding dorsalizing agents to the growth medium. Derivatives of the peripheral nervous system can be generated by inducing the neural crest stem cells to differentiate in vitro.

Abstract: Methods are provided for the establishment and maintenance in long term culture of hormone secreting cells. Cells are derived from tumorous or non-tumorous animal or human tissues, including ovary, endometrium, trophoblast, pituitary, thyroid, and pancreas. The cells secrete into the culture medium hormones such as estrogens, progestins, follicle-stimulating hormone, luteinizing hormone, human chorionic gonadotrophin, thyroxin, glucagon, and insulin, depending on the tissue of origin of individual cell cultures. Contact with an appropriate secretogogue causes the cells to respond with increased hormone secretion. For instance, ovarian follicular cells respond to follicle-stimulating hormone with increased estrogen and progesterone secretion. Pancreatic cells respond to elevated glucose with increased insulin secretion. The cells proliferate in in vitro for up to one year or longer, during which time they retain their hormone-secretion profile.

Abstract: The invention concerns a scaffold which is used as a growth supportive base for various cells and tissue explants from three-dimensional tissue comprising naturally derived connective or skeletal tissue into attached flakes having a very high porosity. Alternatively the scaffold is composed of fused epiphyses.

Abstract: Methods for treating treatment-naive as well as treatment-experienced patients having RCC to achieve at least a partial tumor response involving administering a therapeutically effective amount of pegylated interferon-alpha, e.g., pegylated interferon alpha-2b as monotherapy or in association with a therapeutically effective amount of IL-2 are disclosed.

Abstract: A chemically defined-serum free growth medium for the in vitro and ex vivo of cells and cell lines. The medium consists of about a one to one ratio (v/v) of two basal growth media containing &agr;-ketoglutarate, insulin, transferrin, selenium, bovine serum albumin, linoleic acid, ceruloplasmin, cholesterol, phosphatidyl-ethanolamine, &agr;-tocopherol acid succinate, reduced glutathione, taurine, triiodothyronine, hydrocortisone, parathyroid hormone, L-ascorbic acid 2-sulfate, &bgr;-glycerophosphate, PDGF, EGF and FGF. Chondrocytes, when cultured in this medium in the presence of a cartilage derived morphogenetic protein or bone morphogenetic protein, retain their cartilaginous phenotype.

Abstract: The present invention provides a module for cell culture comprising a structure comprising hollow fibers and at least two spacers, wherein the spacers have small pores regularly arranged therein and the hollow fibers pass through the small pores and are arranged regularly at a very small distance, a hybrid artificial liver module wherein hepatocytes are immobilized in this module, and a method of cell culture wherein cells are immobilized in the lumen or extra-fiber space of the module of this module by the use of a centrifugal force and wherein the immobilized cells are cultured. The present invention also provides a method for cell culture wherein hepatocytes are cultured in a culture medium containing DMEM as a basal medium.

Abstract: The invention includes compositions, kits, and methods for modulating survival and differentiation of mammalian multi-potential hematopoietic progenitor cells using a placental glycoprotein hormone of the murine prolactin family, namely either murine prolactin-like protein E or murine prolactin-like protein F. The compositions, kits, and methods described herein can be used, for example, for in vitro or ex vivo expansion of hematopoietic precursor cells or to treat a disorder associated with aberrant hematopoiesis (e.g., pre-eclampsia and thrombocytopenia).

Abstract: The invention relates to cell proliferation, cell differentiation, male infertility, male fertility and to compositions and methods involved therein. Also methods of culturing spermatogonial stem cells with bone morphogenetic protein 8 are disclosed.

Abstract: A culture medium for culturing chicken embryonic stem (ES) cells is disclosed. The culture medium is used for supporting avian ES cell cultures. The components of the avian ES cell media include cytokines, fibroblast growth factors, insulin-like growth factors and stem cell growth factors and anti-retinoic acid antibody. The culture medium is substantially free of active retinoic acid. A method for culturing avian ES cells and the resulting products are also disclosed.

Abstract: Isolation, characterization, proliferation, differentiation and transplantation of mammalian neural stem cells is disclosed.

Abstract: 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-1771) 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.

Abstract: The present invention provides a method of treating cancer comprising (a) obtaining a tumor cell line, (b) modifying the tumor cell line to render it capable of producing an increased level of a cytokine relative to the unmodified tumor cell line, and (c) administering the tumor cell line to a mammalian host having at least one tumor that is the same type of tumor as that from which the tumor cell line was obtained, wherein the tumor cell line is allogeneic and is not MHC-matched to the host. The present invention also provides a pancreatic tumor cell line, a method and medium for obtaining such a tumor cell line, and a composition comprised of cells of a purified pancreatic tumor cell line.

Abstract: A chemically defined mammalian cell culture medium is provided that supports maintenance and long term clonal growth of mammalian hepatocytes and other cells.

Abstract: A method is disclosed for the in vitro growth and proliferation of germ cells obtained from the ovarioles of an insect. By culturing these cells in a medium supplemented with soluble cytokines and mitogenic agents and independent of feeder-cells, they can be induced to proliferate. Cells are stored by cryogenic means for future use.

Abstract: A method for producing a neuroblast and a cellular composition comprising an enriched population of neuroblast cells is provided. Also disclosed are methods for identifying compositions which affect neuroblasts and for treating a subject with a neuronal disorder, and a culture system for the production and maintenance of neuroblasts.

Abstract: The present invention provides a cell culture medium useful for a biochemical analysis of antioxidant function in human lymphocytes, said medium comprising, a buffered, serum-free solution containing the following ingredients: a carbohydrate selected from the group consisting of glucose and a compound biologically capable of producing glucose in the cells, a biologically usable form of pantothenic acid, choline or a biological usable form of a substance capable of producing choline in the cells, inorganic ions comprising chloride, phosphate, calcium, magnesium, potassium, sodium, and iron in a biologically utilizable form, cumene hydroperoxide, deionized water, and a mitogen in an amount effective to stimulate the lymphocytes being assayed; said buffered, serum-free solution having a pH from about 6.8 to 7.6, said cell culture medium characterized by being effective to determine nutritional deficiencies, inadequacies, and imbalances and to biochemically analyze antioxidant function of the lymphocytes.

Abstract: Methods and compositions are provided for in vitro expansion of growth factor dependent cells. Expansion is effected through the use of growth factor conjugates that include a growth factor such as a steel factor and a polysaccharidase substrate binding region. The conjugates are immobilized by binding of the substrate binding region to a substrate of the polysaccharidase in a growth chamber for the cells.

Abstract: Human pancreatic endocrine cells are proliferated without loss of hormone function in a culture medium containing extracellular matrix from bladder carcinoma cell lines in the substantial absence of hepatocyte growth factor/scatter factor. Proliferation is preferably carried out in the substantial absence of any peptide growth factors and nicotinamide. The cells may be proliferated in a monolayer on a solid substrate. Islets and islet-like cell clusters are proliferated without loss of insulin-secreting function by incubation in a medium containing extracellular matrix from a human bladder carcinoma cell line, preferably cell line ATCC HTB-9.

Abstract: The present invention relates to a substantially pure population of viable bile duct progenitor cells, and methods for isolating such cells. The present invention further concerns certain therapeutic uses for such progenitor cells, and their progeny.

Abstract: A method for enhancing the survival and/or proliferation of Schwann cells (especially human Schwann cells) in cell culture is disclosed which involves culturing the cells in serum free culture medium comprising gas6 and other mitogenic agents, such as heregulin and forskolin. The culturing step is generally preceded by a pre-incubation period wherein nerve tissue comprising the Schwann cells is cultured under appropriate conditions and for a period of time such that demyelination occurs. The isolated Schwann cells can be used as cellular prostheses to treat patients with nervous system injuries. The invention also provides a cell culture medium for culturing Schwann cells.

Abstract: In vitro production of clinically useful quantities of single species of mature, differentiated human blood cells is carried out by a method in which human pluripotent hematopoietic stem cells, preferably from a universal donor, are incubated in a bioreactor in a growth medium also containing specific combinations of recombinant human growth and maturation promoting polypeptide factors that expand stem cell cultures and promote the maturation and differentiation of stem cells into single species of erythroid, thrombocytic or granulocytic human blood cells, and harvesting the mature cells. The growth and maturation promoting polypeptides employed include SCGF, Interleukins 1,3,4,5,6, and 11, GM-CSF, M-CSF, G-CSF and EPO. Stem cells may be preliminarily genetically modified so as to remove histocompatibility or blood group antigens with which a recipient may be incompatible, or the stem cells may be genetically altered by transfection with appropriate DNA-containing vectors, prior to addition to the bioreactor.

Abstract: Disclosed is a synthetic medium with PDGF, vitronectin, IL-1.beta. and BSA added to Eagle's minimum essential medium or medium with transferrin, insulin, progesterone and putrescine further added thereto. When cultivating the postnatal central neurons using the inventive medium, there are effects such that good attachment to substrate, extension of neuritic processes and maintenance of survival are achieved, that more stable sure cultivation becomes possible as well over the astrocyte-conditioned medium used hitherto, and the like.

Abstract: A method of culturing non-transformed mammalian respiratory epithelial cells in vitro. The method includes the steps of harvesting the epithelial cells from the respiratory tract of a mammalian donor; suspending the epithelial cells in a fluid culture medium to form a cell suspension; introducing the cell suspension into a first reservoir in communication with an adherence substrate; introducing additional fluid culture medium into a second reservoir separated from the first reservoir by a fluid permeable baffle; incubating the cells under conditions suitable for promoting epithelial cell growth for a period of at least 72 hours; and changing the fluid culture medium within the second reservoir during the incubation period at intervals of approximately 24-72 hours without disturbing the epithelial cells within the first reservoir. Preferably, the epithelial cells are harvested from the nasal mucosa of a human donor using a minimally-invasive brushing technique.

Abstract: A method for enhancing the survival and/or proliferation of Schwann cells (especially human Schwann cells) in cell culture is disclosed which involves culturing the cells in serum free culture medium comprising gas6 and other mitogenic agents, such as heregulin and forskolin. The culturing step is generally preceded by a pre-incubation period wherein nerve tissue comprising the Schwann cells is cultured under appropriate conditions and for a period of time such that demyelination occurs. The isolated Schwann cells can be used as cellular prostheses to treat patients with nervous system injuries. The invention also provides a cell culture medium for culturing Schwann cells.

Abstract: A biochemically defined culture medium for culturing engineered Chinese hamster ovary (CHO) cell lines, which is essentially free from protein, lipid and carbohydrate isolated from an animal source, having water, an osmolality regulator, a buffer, an energy source, amino acids including L-glutamine, an inorganic or recombinant iron source, and a synthetic or recombinant growth factor, and optionally non-ferrous metal ions vitamins and cofactors. Also cells adapted to grow in such a culture medium.

Abstract: The present invention relates to monoclonal antibodies having high affinity to human pancreatic PLA.sub.2, the production thereof, hybridomas producing them and an immunoassay for human pancreatic PLA.sub.2 using them.

Abstract: A biochemically defined culture medium for culturing engineered Chinese hamster ovary (CHO) cell lines, which is essentially free from protein, lipid and carbohydrate isolated from an animal source, having water, an osmolality regulator, a buffer, an energy source, amino acids including L-glutamine, an inorganic or recombinant iron source, and a synthetic or recombinant growth factor, and optionally non-ferrous metal ions vitamins and cofactors. Also cells adapted to grow in such a culture medium. REEXAMINATION RESULTS The questions raised in reexamination request no. 90/006656, filed Jun. 2, 2003 have been considered and the results thereof are reflected in this reissue patent which constitutes the reexamination certificate required by 35 U.S.C. 307 as provided in 37 CFR 1.570(e), for ex parte reexaminations, or the reexamination certificate required by 35 U.S.C. 316 as provided in 37 CFR 1.997(e) for inter partes reexaminations.