Abstract: The present invention provides neuronal progenitor cells which have been identified in histological sections of the adult human brain. The present invention also provides methods to localize, characterize, harvest, and propagate neuronal progenitor cells derived from human brain tissue. Additional methods are provided for introducing and expressing genes in the brain.

Abstract: The present invention provides a method for enhancing the immunogenicity of weakly immunogenic or non-immunogenic cells, resulting in a cellular vaccine that can stimulate T cell activation, which in turn leads to an effective immune response. The cellular vaccines of the present invention are useful for the prevention and treatment of diseases which develop and/or persist by escaping the immune response triggered by T cell activation. Such diseases include, for example, all cancers, natural and induced immune deficiency states, and diseases caused by infections with a variety of pathogens.

Abstract: Methods of production of desired recombinant proteins, polypeptides and peptides are disclosed which utilize mammalian host cells engineered for autonomous and regulated growth in low cost protein/serum-free culture media. Preferred host cells express insulin or an insulin-like growth factor, and/or transferrin and are engineered so that addition of an inducer (e.g., ZnCl2+CdCl2) halts growth and simultaneously induces the expression of the desired recombinant protein, polypeptide or peptide.

Abstract: The subject invention pertains to compositions and methods for culturing nerve tissue in vitro and nerve grafts produced using such methods. The compositions and methods of the subject invention can be employed to restore the continuity of nerve interrupted by disease, traumatic events or surgical procedures. The invention also concerns methods for promoting repair of damaged nerve tissue using the present compositions and nerve tissue treated according to such methods.

Abstract: The invention provides a method of culturing cells which includes a proliferating step in which the number of precursor cells is expanded and a differentiating step in which the expanded precursor cells develop into neuronal cells. The proliferating step includes the step of incubating the precursor cells in proliferating medium which includes basic fibroblast growth factor (bFGF). The differentiating step includes incubating the precursor cells in differentiation media in a manner effective to form a cellular aggregate that is not adhered to any surface of the incubation vessel. In a preferred embodiment, the cells arc incubated in a roller tube. The differentiation media can also include at least one differentiating agent. The invention also provides a method for treating a neurological disorder, such as Parkinson's disease, a method of introducing a gene product into a brain of a patient, an assay for neurologically active substances, and a cell culture.

Abstract: Methods of treating and forming biocompatible microcapsules that contain living cells are provided, to improve the function of the microcapsules. In particular, methods of treating islet cells or microcapsules containing islet cells are provided. Culture of isolated islet cells prior to encapsulation, culture of encapsulated cells, and cryopreservation of islet cells prior to encapsulation, are described. Methods for harvesting viable islets that incorporates an anti-oxidant in the digestion medium are also disclosed.

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: The present invention provides cell culture media formulations which support the in vitro cultivation of animal epithelial cells. The media comprise at least one fibroblast growth factor (FGF) and at least one agent that induces increased intracellular cAMP levels, and optionally comprise ascorbic acid. The present invention also provides methods of cultivating animal epithelial cells in vitro using these cell culture media formulations, kits comprising the media, cell culture compositions comprising the culture media and an animal epithelial cell, and compositions that may be used as replacements for organ or gland extracts in animal cell culture media.

Abstract: A chondrogenesis promoter comprising as an active ingredient a compound represented by general formula (I) or a salt thereof: wherein R1 represents a halogen atom, a lower alkyl group, a lower alkoxy group, etc.; R2 represents a hydrogen atom, a lower alkyl group, a lower alkenyl group, a lower alkynyl group, a lower alkoxy group, an acyl group, an aryl group, a heterocyclic group, etc.; R3 represents a lower alkyl group, a cycloalkyl group, an aryl group, a heterocyclic group, etc.; R4 represents a hydrogen atom, a lower alkyl group, an aryl group, a heterocyclic group, etc.; X and Y represent —CH2—, —NH— or —O—; and n represents an integer of 0-4.

Abstract: Methods, including culture media conditions, which provide for in vitro human stem cell division and/or the optimization of human hematopoietic progenitor cell cultures and/or increasing the metabolism or GM-CSF secretion or IL-6 secretion of human stromal cells and/or a method for assaying the effect of a substance or condition on a human hematopoietic cell population, and/or depleting the malignant cell or T-cell and B-cell content of a human hematopoietic cell population are disclosed. The methods rely on culturing human stem cells and/or human hematopoietic progenitor cells and/or human stromal cells in a liquid culture medium which is replaced, preferably perfused, either continuously or periodically, at a rate of 1 ml of medium per ml of culture per about 24 to about 48 hour period, and removing metabolic products and replenishing depleted nutrients while maintaining the culture under physiologically acceptable conditions. Optionally, growth factors are added to the culture medium.

Abstract: A novel co-culture system using human brain endothelial cells (HUBEC) which promotes the expansion of human CD34+CD38− cells consistent with the PMVEC system is disclosed. HUBEC were isolated from cadaveric donors, passed in primary culture, cloned and found to be Von Willebrand Factor positive. Cultivation of purified bone marrow CD34+ cells on HUBEC monolayers supplemented with GM-CSF+IL-3+IL-6+SCF+flt-3 ligand caused a 14.5-fold increase in total cells, an 6.6-fold increase in CD34+ cells, and, most remarkably, a 440-fold increase in CD34+CD38− cells after 7 days. Further, CFU-GM production increased 15.1-fold, BFU-E increased 8-fold, and CFU-Mix increased 5.2-fold. Optimal generation was dependent upon the continued presence of exogenous supplied cytokines. Moreover, we found that non-brain human endothelial cells isolated from the same donors supported neither the expansion nor the maintenance of human CD34+CD38− cells.

Abstract: This invention provides media that support the growth of primate pluripotent stem cells in feeder-free culture, and cell lines useful for producing such media and other purposes. Conventionally, it has been necessary to grow pluripotent embryonic cells on feeder layers of primary embryonic fibroblasts, in order to prevent them from differentiating. It has now been discovered that standard culture media conditioned by special cell lines can be used to support proliferation of pluripotent stem cells while inhibiting differentiation in an environment free of feeder cells. This invention includes mesenchymal and fibroblast-like cell lines obtained from embryonic tissue or differentiated from embryonic stem cells. Methods for deriving such cell lines, processing media, and growing stem cells using the conditioned media are described and illustrated in this disclosure.

Abstract: Using a novel culture approach, previously unknown populations of neural progenitor cells have been found within an adult mammalian brain. By limiting cell-cell contact, dissociated adult brain yields at least two types of cell aggregates. These aggregates or clones of stem/precursor cells can be generated from adult brain tissue with significantly long postmortem intervals. Both neurons and glia arise from stem/precursor cells of these cultures, and the cells can survive transplantation to the adult mammalian brain.

Abstract: Provided are methods and compositions for increasing the efficiency of nuclear transfer using apoptosis inhibitors, and for the production of transgenic and non-transgenic mammals from cultured cells or cell lines. Methods for cloning mammals, and for producing transgenic and chimeric mammalian tissues and mammals, and chimeric cell lines are also provided.

Abstract: A monolayer sheet structure is prepared containing a confluent monolayer of primary hepatocytes cultured on a substrate in sheet form. The monolayer sheet structure is formed by treating the hepatocytes in the monolayer formation stage with a protein-phosphorylation inhibitor having an indolo [2,3-a] carbazole structure, such as staurosporine, or other similar structure.

Abstract: Tissue engineering is the development of biological substitutes to restore, maintain, or improve tissue function. One strategy that has been created to regenerate new tissue comprising the steps of providing cells, expanding the isolated cells in a first culture medium in which the cells lack differentiated functions and redifferentiating the expanded cells in a second cell culture medium. The present invention provides an improved method for tissue engineering. In particular, the method provides specific biochemical factors to supplement cell culture medium during the redifferentiation process with the goal of regenerating tissue equivalents that resemble natural tissues both structurally and functionally. These specific chemical factors induce and/or accelerate and/or promote the redifferentiation of the previously expanded cells.

Abstract: The present invention provides a method for ameliorating &bgr;-globin disorders in a mammal. In one aspect of the invention, the treatment involves ex vivo treatment of early erythroid progenitor cells that leads to an increase in the relative amounts of cells subsequently expressing and accumulating HbF. The cell treatment is to be followed by transplantation of the modified cells. In another aspect of the invention, the same modification of progenitor cells occurs in vivo. Both treatments are based on the novel discovery that the modification can be performed very early in the erythroid maturation process, without disturbance of the subsequent proliferation and maturation of the erythroid precursor. The present invention also provides a procedure for the monitoring of &bgr;-globinopathies and the response of a patient to treatment.

Abstract: The invention relates to compositions for in vitro fertilization, or for the in vitro culture of follicles, male germinal cells or embryos, said compositions comprising at least two growth factors in association with a compound of the family of corticoids involved in energetic production in mammals. The invention also relates to the culture media obtained with these compositions, these latter being preferably in the form of a lyophilizate. The invention also has for its object processes for the in vitro fertilization and processes for maturation of follicles, of male germinal cells, or embryos using such culture media, as well as kits for the practice of these processes and comprising a lyophilizate as mentioned above and an aqueous solution, if desired commercial, containing other constituents of said culture medium.

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: Disclosed are compositions for mammalian, avian or piscian reproductive cells and methods for the collection, holding, processing, in vitro fertilization, sexing culturing, or storing (including long-term cryopreservation) of mammalian, avian, or piscian reproductive sperm cells. The compositions comprise a suitable reproductive cell media and a transforming growth factor, an insulin-like growth factor, or zinc, and, optionally, inositol, transferrin, or fructose, or combinations thereof.

Abstract: Disclosed is a medium for the propagation of early stage embryos to blastocyst stage. The medium contains an effective amount of GM-CSF to increase the percentage of pre-blastocyst embryos which develop to transfer ready blastocysts. Also disclosed is a method of growing early stage human embryos to transfer ready blastocysts. The method includes the step of incubating the embryos in vitro in a culture medium containing an effective amount of human GM-CSF for a time and under conditions to increase the proportion of transfer ready blastocysts. An IVF program that includes the method of growing early stage human embryos to transfer ready blastocysts is also disclosed.

Abstract: Production of fully differentiated, optionally immortalized, neural cells—by enhancing replication then inducing differentiation by mimicking cell's natural environment in vitro. The cells are useful for transplantation or drug screening.

Abstract: A process of using a fish plasma component as a nutrient medium component for tissue culture includes obtaining blood from a fish, separating plasma from the blood, and extracting one or more specific components of the plasma. The tissue is cultured using the extracted plasma components, and none of any remainder of the plasma, in a nutrient medium. The tissue cultured using the extracted plasma component is other than fish tissue, such as mammalian tissue or insect tissue.

Abstract: A novel medium for maintaining and growing pluripotent and germ line competent mammalian stem cells. The medium is conditioned by a fibroblast cell clone that produces leukemia inhibitory factor. The cells are placed in the medium so that, over a period of time, the leukemia inhibitory factor accumulates in the medium. A portion of the medium is then removed, and the quantity removed is replaced with unconditioned medium.

Abstract: A method is described whereby dendritic cells derived from the CD34+ and CD 34− hematopoietic cell lineages are directed to become programmable antigen presenting cells. The programmed cells may be pulsed with tumor cell RNA or tumor cell RNA expression products. The protocol provides for directing the maturation of dendritic cells to become antigen presenting cells. The protocol further provides for isolating tumor cell RNA from biopsy material that has been prepared in paraffin block storage. The directed dendritic cell is provided with a plurality of tumor markers by using tumor RNA in toto, the poly A+RNA fraction or the expression product of such RNA. Once activated the dendritic cells are incubated with T4 and T8 lymphocytes to stimulate and sensitize the T lymphocytes which upon introduction either into a donor host or a nondonor recipient will provide immune response protection.

Abstract: A culture system for producing PGCs or EG cells by culturing PGCs for long periods in tissue culture is provided. This culture system uses LIF, bFGF, IGF and SCF. The resultant EG cells are useful for the production of transgenic and chimeric avians, in particular, chickens and turkeys, and also for cloning purposes.

Abstract: The present invention is directed generally to cell culture media (particularly serum free, non animal derived, and/or chemically defined media) which are useful for introducing macromolecules and compounds (e.g., nucleic acid molecules) into cells (e.g., eukaryotic cells). According to the invention, such introduction can take place in the presence of said medium. Cells containing such introduced materials can then be cultured in the medium and the effect of the introduced materials on the cells can be measured or determined. In particular, the invention allows introduction of nucleic acid molecules (e.g., vectors) into cells (particularly eukaryotic cells) and expression of proteins encoded by the nucleic acid molecules in the cells. The invention obviates the need to change the cell culture medium each time a different procedure is performed with the cells (e.g., culturing cells vs. transfecting cells).

Abstract: The nerve growth factor (NGF) is used for the storage of corneas in culture, for the production and the storage in vitro of single cell populations of the corneal morphological and functional unit (i.e. epithelium, stroma/keratocytes and endothelium) and of the conjunctival epithelium, and for the production and the storage of corneal and conjunctival tissues, in particular for transplantation purposes. The NGF is also proposed for use in the therapy and/or the prophylaxis of diseases of the corneal surface, wherein a lack of integrity of the corneal and conjunctival morphological and functional unit occurs, in particular for pathologies having a dystrophic or neurodystrophic basis, both congenital and acquired.

Abstract: A method of inducing the maturation of dendritic cells by stimulating immature dendritic cells with an imidazoquinoline type immune response modifying compound. Dendritic cells that have been matured in this manner display increased antigen presenting ability and may be used as immunotherapeutic agents.

Abstract: Methods and compositions for treating diabetes mellitus in a patient in need thereof are provided. The methods include administering to a patient a composition providing a gastrin/CCK receptor ligand, e.g., a gastrin, and/or an epidermal growth factor (EGF) receptor ligand, e.g., TGF-&agr;, in an amount sufficient to effect differentiation of pancreatic islet precursor cells to mature insulin-secreting cells. The composition can be administered systemically or expressed in situ by cells transgenically supplemented with one or both of a gastrin/CCK receptor ligand gene, e.g., a preprogastrin peptide precursor gene and an EGF receptor ligand gene, e.g., a TGF-&agr; gene. The methods also include transplanting into a patient cultured pancreatic islets in which mature insulin-secreting beta cells are proliferated by exposure to a gastrin/CCK receptor ligand and an EGF receptor ligand.

Abstract: Disclosed is a method of stimulating adult corneal endothelial cell proliferation, which includes incubation of corneal endothelium to a mitogen-containing medium and then to an agent that interrupts cell-cell contacts in the adult corneal endothelial cells, and subsequently, as needed, further incubation in the mitogen-containing medium. The cell-cell contact-interrupting agent is preferably a calcium chelator.

Abstract: The nerve growth factor (NGF) is used for the storage of corneas in culture, for the production and the storage in vitro of single cell populations of the corneal morphological and functional unit (i.e., epithelium, stroma/keratocytes and endothelium) and of the conjunctival epithelium, and for the production and the storage of corneal and conjunctival tissues, in particular for transplantation purposes. The NGF is also proposed for use in the therapy and/or the prophylaxis of diseases of the corneal surface, wherein a lack of integrity of the corneal and conjunctival morphological and functional unit occurs, in particular for pathologies having a dystrophic or neurodystrophic basis, both congenital and acquired.

Abstract: The method for obtaining and expanding TcR&ggr;&dgr;+ T cells in culture is described. The method involves: 1) culturing cells from a sample containing TcR&ggr;&dgr;+ T cells or precursors thereof in a first culture medium comprising a T cell mitogen and at least two cytokines and 2) culturing the cells obtained in step 1) in a second culture medium comprising at least two cytolines. Preferably, the method comprises 1) culturing the cells in a first culture medium comprising (a) a T cell mitogen, (b) interleukin-2 and (c) interleukin-4, and 2) culturing the cells obtained in step 1) in a second culture medium comprising (i) interleukin-2 and (ii) interleukin-4 to obtain TcR&ggr;&dgr;+ T cells. The TcR&ggr;&dgr;+ T cells obtained by the method can be used in a variety of experimental, therapeutic and commercial applications.

Abstract: The stem cell-augmenting material of the present invention being capable of augmenting stem cells consisting of somatic stem cells, particularly hematopoietic stem cells of bone marrow, and embryonic stem cells by being absorbed in vivo by oral ingestion, drops, etc. and capable of augmenting stem cells using isoflavone aglycone, which has estrogen-like activity and does not block the enzyme activity of enzymes that act on cell proliferation factor. There have been no stem cell-augmenting materials like the present invention whatsoever. The main characteristic of the present invention is that a stem cell-augmenting material that has the excellent activity is obtained because it has isoflavone aglycone that has a stem cell-augmenting activity.

Abstract: Nucleic acids may be obtained from neural cell cultures produced by using growth factors to induce the proliferation of multipotent neural stem cells. The resultant progeny may be passaged repeatedly to produce a sufficient number of cells to obtain representative nucleic acid samples. Clonal cultures may be produced. Nucleic acids may be obtained from both cultured normal and dysfunctional neural cells and from neural cell cultures at various stages of development. This information allows for the identification of the sequence of gene expression during neural development and can be used to reveal the effects of biological agents on gene expression in neural cells. Additionally, nucleic acids derived from dysfunctional tissue can be compared with that of normal tissue to identify genetic material which may be the cause of the dysfunction. This information could then be used in the design of therapies to treat the neurological disorder.

Abstract: Cell based screens for studying drug transport are described and improved methods for the preparation of cell monolayers for use in such screens are disclosed.

Abstract: A method for culturing and/or multiplying lymphocytes in cell culture medium which contains as lymphocyte growth factor and additionally, aurin tricarboxylic acid, ciclosporin, tacrolimus, and/or ascomycin.

Abstract: The invention relates to a culture medium for culturing cells, in particular human cells in a process for tissue engineering bone. The medium comprises glucose, a mineral, a vitamin, a growth factor and L-glutamine, wherein the L-glutamine is present in a concentration of at least 300 mg/L.

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: A scalable method for the production of highly purified plasmid DNA in Escherichia coli is described, which method includes growing plasmid-containing cells to a high biomass in exponential growth and lysing the cells by raising the pH of the culture to a carefully controlled pH value in which chromosomal DNA is denatured but plasmid DNA is reversibly renatured. The method has been developed for the production of pharmaceutical grade DNA for use in in vivo and ex vivo gene therapy.

Abstract: A culture medium for avian embryonic cells and an avian cell culture medium is disclosed. The culture medium is characterized in that it has elements complementary to avian embryonic cells. The complementary elements are cytokines, fibroblast growth factors, insulin-like growth factors or stem cell growth factors. The medium is substantially free of active retinoic acid. A method for culturing avian embryonic cells, and the resulting products, are also disclosed.

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: A culture medium for avian totipotential embryonic cells comprising an avian cell culture medium is disclosed. The culture medium is characterised in that it comprises elements complementary to said avian cell culture medium, the complementary elements being selected from the group which comprises cytokines, fibroblast growth factors, insulin-like growth factors and stem cell growth factors, and in that it is substantially free of active retinoic acid. A method for culturing avian totipotential embryonic cells, and the resulting products, are disclosed.

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. The culturing process is conducted for an extended period of time, so that any latent red blood cells are eliminated from the islet culture.

Abstract: An embryonic stem cell which may be induced to differentiate homogeneously into a desired primary cell line. The embryonic stem cell may be engineered with DNA, which encodes a protein or polypeptide which promotes differentiation of the stem cell into a specific cell line, such as, for example, a neuronal cell line, a muscle cell line, or a hematopoietic cell line. The DNA may encode a transcription factor found in the particular cell line. In another alternative, a desired cell line is produced from embryonic stem cells by culturing embryonic stem cells under conditions which provide for a three-dimensional network of embryonic stem cells, and then stimulating embryonic stem cells with an agent, such as retinoic acid, or dimethylsulfoxide, which promotes differentiation of the embryonic stem cells into the desired cell line, such as, for example, a neuronal cell line, or a muscle cell line.

Abstract: Compositions containing clinically relevant numbers of immune cells that have been isolated from a patient differentiated and/or expanded ex vivo. Methods for treating or preventing disease or otherwise altering the immune status of the patient by reinfusing such cells into the donor are also provided. Methods for expanding and/or immune cells, including effector cells, in the absence of exogenous IL-2, and for administering the cells in the absence of co-infused IL-2 are also provided.

Abstract: The present invention features methods of modulating primary stem cell differentiation in culture by altering the endogenous activity of an insulin-like growth factor.

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: The use of professional antigen presenting cells genetically modified to enhance expression of an immunostimulatory cytokine is disclosed for the treatment of individuals having tumors or infections. The genetically modified professional antigen presenting cells are injected directly at or near the site of the tumor or infection. Preferred professional antigen presenting cells include dendritic cells, and preferred immunostimulatory cytokines include interleukins such as IL-12.

Abstract: The invention features methods for expansion and packaging of cells. Expansion is achieved by providing cells that include neural progenitor cells; plating the cells in culture vessels at an average density of 1×105 to 7×105 cells/cm2; and culturing said cells in culture medium and under conditions permissible for proliferation of said neural progenitor cells, wherein the volume of medium results in an initial cell density of between 5×104 and 1.5×105 cells per milliliter of medium. Preparation of cells for transplantation includes providing a cell suspension that includes single cells, aggregates of fewer than two hundred cells, or a combination thereof; and re-aggregating the cells in said cell suspension, wherein greater than 50% of reaggregates consist of between 25 and 500 cells/reaggregate.