Abstract: The invention relates to stem cells isolated from the retina of mammals and retinal cells differentiated from these stem cells. The invention also relates to a method of isolating retinal stem cells and inducing retinal stem cells to produce retinal cells. Retinal stem cells may also be induced in vivo to produce retinal cells. The invention also includes pharmaceuticals made with retinal stem cells or retinal cells which may be used to restore vision lost due to diseases, disorders or abnormal physical states of the retina. The invention includes retinal stem cell and retinal cell culture systems for toxicological assays, for isolating genes involved in retinal differentiation or for developing tumor cell lines.

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: Compounds termed "vitaletheine modulators" which include beta-alanyl-taurine and carbobenzoxy beta-alanyl-taurine are synthesized and added to culture media for in vitro culture of cells such as mammalian or plant cells. The compounds support cell vitality, and provide increased cellular life span, increased cellular bioproductivity, improved cellular function, and adaption of resistant cells to culturing. Carbobenzoxy beta-alanyl-taurine is produced by coupling .beta.-alanine, which has it's terminal amine protected with a carbobenzoxy (CBZ) group, to N-hydroxysuccinimide to produce an active ester of .beta.-alanine, coupling two of the active esters to cystamine to produce a CBZ-protected .beta.-alethine having an internal disulfide bond, isolating and purifying the CBZ-protected .beta.-alethine, and reacting the CBZ-protected .beta.-alethine with iodine to oxidize the disulfide bond to obtain carbobenzoxy beta-alanyl-taurine.

Abstract: Methods of obtaining enriched populations of osteoclast precursor cells which can be released from tissue culture dishes and used for biochemical studies are described. Osteoblastic cells and bone marrow cells are co-cultured. Next a .alpha..sub.v .beta..sub.3 receptor ligand, such as echistatin is used for cell detachment. The result is a 75-95% pure enriched population of tartrate resistant acid phosphatase (TRAP.sup.+) cells, in high yields (2-3.times.10.sup.6 cells per experiment) can be obtained. These cells are mosty mononucleated and based on their characteristics are considered to be pre-fusion osteoclasts (pOC cells). The precursor osteoclasts can be reseeded onto osteoblasts to obtain an enriched population of mature, multinucleated osteoclast cells.

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: Methods are provided for eluting peptides that are bound to major histocompatibility complex ("MHC") molecules expressed on the cell surfaces of viable cells that have at least one MHC-peptide complex on the surfaces of the cells. Methods are provided for using such acid-eluted T cell epitopes, preferably obtained from a patient's tumor, and autologous dendritic cells as the basis for antitumor vaccines.

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: The present invention provides a rapid expansion method (termed "REM"), for quickly generating large numbers of T lymphocytes, including cytolytic and helper T lymphocytes. REM involves culturing the T cells in association with a disproportionately large concentration of nondividing feeder cells, preferably .gamma.-irradiated peripheral blood mononuclear cells ("PBMC") present at an excess of at least 40-fold (relative to the number of target T cells), more preferably at an excess of at least about 200-fold. Cultures grown under REM exhibit dramatically enhanced expansion rates that can be even further elevated by the use of appropriate concentrations of an additional feeder cell, an anti-CD3 monoclonal antibody and IL-2, as described herein. Clonal expansions in the range of 500-fold to 3000-fold can be achieved within a single stimulation cycle of about 10-13 days, which is more than 100-fold more efficient than currently employed methods of culturing human T cell clones.

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 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 perfusion device such as a liver assist device containing a housing defining a perfusion inlet and a perfusion outlet, a porous membrane structure mounted within said housing to define a perfusion compartment and an adjacent hepatocyte compartment, and porcine hepatocytes isolated from a porcine liver by retrograde perfusion.

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 subject invention concerns new methods which make it possible, for the first time, to grow functional islets in in vitro cultures. The subject invention also concerns the use of the in vitro grown islet-like structures for implantation into a mammal for in vivo therapy of diabetes. The subject invention further concerns a process using the in vitro grown islet implants for growing an organ in vivo that has the same functional, morphological and histological characteristics as those observed in normal pancreatic tissue. The ability to grow these cells in vitro and organs in vivo opens up important new avenues for research and therapy relating to diabetes.

Abstract: A method for stimulating chondrocyte proliferation and inhibiting chondrocyte differentiation along the endochondral developmental pathway is provided comprising contacting condrocytes with an effective amount of Platelet-Derived Growth Factor (PDGF) such as PDGF-BB, PDGF-AA OR PDGF-AB in the substantial absence of growth factors which promote cell differentiation. This allows such cells to be multiplied in culture for loading onto a scaffolding material and implanting into a cartilage or bone wound.

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: The invention relates to methods of enriching for hematopoietic cell populations enriched in myeloid and/or lymphoid progenitor cells based on cell specific markers. The methods also provide an enriched population of prethymic lymphoid-committed progenitor population lacking long-term hematopoietic reconstitution potential. Compositions enriched for the cells and populations of cells obtained therefrom are also provided by the invention. Methods of use of the cells are also included. Methods of genetically modifying the cells are provided as are cells obtained thereby.

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 method is described for inducing in vivo proliferation of precursor cells located in mammalian neural tissue by administering to the mammal a fibroblast growth factor and at least one additional growth factor selected from the group consisting of epidermal growth factor, transforming growth factor alpha, and amphiregulin. The method can be used to replace damaged or missing neurons and/or glia. Another method is described for transplanting multipotent neural stem cell progeny into a mammal. The method comprises the steps of administering growth factors to a mammal to induce in vivo proliferation of neural precursor cells, removing the precursor cell progeny from the mammal, culturing the removed cells in vitro in the presence of one or more growth factors that induces multipotent neural stem cell proliferation, and implanting the multipotent neural stem cell progeny into the mammal.

Abstract: The present invention relates to the treatment of skin defect by organotypically cultured autologous keratinocytes isolated from the outer root sheath of hair follicles. Methods for primary as well as subsequent organotypic cultures (epidermal equivalents) in fully defined media eventually supplemented by autologous human serum and substances isolated from blood components, with minimal allogeneic biological supplements, are disclosed. Techniques to prepare epidermal equivalents for transplantation are included, as well as a method for the transport of the epidermal equivalents.

Abstract: The invention features a method for the selection and expansion of bone marrow stromal cells. The method includes the steps of obtaining bone marrow stromal cells; introducing the stromal cells into a vessel pre-coated on an inner surface with a gelatin, and containing a culture medium including an acidic fibroblast growth factor ("aFGF") polypeptide; and expanding the stromal cells in the culture medium under conditions and for a time sufficient to obtain an increased number of bone marrow stromal cells. The culture medium additionally can include heparin, and the vessel additionally can be precoated with fetal bovine serum.

Abstract: A method is disclosed for gene transfer into a culture of primitive stem cells which comprises a prestimulation step of adding a blocking agent to block at least one inhibitor of a cell cycle of said primitive stem cells. The prestimulation is time-limited for a period of less than approximately 36 hours so that said culture of primitive stem cells retains hematopoietic potential.

Abstract: A serum-free medium which supports the proliferation and differentiation of CD34.sup.+ cells purified from normal bone marrow, peripheral blood of patients treated with cytokines, and umbilical cord blood is described. The recipe for the formulation is given, which provides a medium suitable for the proliferation and differentiation of CD34.sup.+ cells for use in human therapeutic protocols.

Abstract: The present invention relates to novel pluripotent embryonic cell populations derived from embryonic stem cell populations and methods to produce such pluripotent embryonic cell populations. Disclosed is an embryonic stem cell-derived pluripotent embryoid body cell population having one or more cells capable of developing into cells of hematopoietic and/or endothelial lineage. Also disclosed is an embryoid body cell population-derived mixed population of endothelial and erythroid cells. Also disclosed is an embryoid body cell population-derived embryonic blast cell population capable of developing into a variety of hematopoietic cell types. The invention is additionally directed to embryonic stem cell population-derived T and B cell populations. Methods to identify embryonic cell compounds are also disclosed for therapeutic and experimental use.

Abstract: Media and methods are disclosed for the in vitro formation of a histologically complete human epithelium. The media are serum-free, companion cell or feeder layer free and organotypic, matrix free solutions for the isolation and cultivation of clonally competent basal epithelial cells. The media and methods of the invention are useful in the production of epithelial tissues such as epidermis, cornea, gingiva and ureter.

Abstract: A composition and method for maintaining the viability of human mesenchymal precursor cells in a serum-free environment which composition includes (1) a minimum essential medium; (2) serum albumin; (3) an iron source; (4) insulin or an insulin-like growth factor; and (5) at least one amino acid selected from the group consisting of glutamine, arginine and and cysteine, and is free of serum. Also, a composition and method for culture expanding human mesenchymal precursor cells in a serum-free environment. This composition further includes a mitogen, paricularly a serotonergic agonist. The cells are preferably isolated human mesenchymal stem cells.

Abstract: The effect of GDNF on kidney morphogenesis is disclosed. Methods for stimulating budding and branching of the ureteric epithelium, for stimulating axonal outgrowth, for maintaining ureteric epithelial cells in culture, for preventing apoptosis of ureteric epithelial cells, and for treating diseases using GDNF are also disclosed.

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: 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: An assay for identifying compounds with antiandrogenic activity employing a hamster ductus deferens cell line (DDT1) ATCC CRL-1701 and ATCC CRL-12051 is disclosed, as well as antiandrogenic compounds identified from this assay, particularly the nonsteroidal compound N-(4-chlorophenyl)-(Z,Z)-2,3-bis(cyclopropylmethylene)cyclopentane carboxamide of structural formula (I): ##STR1## This compound is an antiandrogen useful in the treatment and prevention of diseases of the prostate including prostatitis, benign prostatic hyperplasia (BPH) and prostatic carcinoma.

Abstract: The present invention relates to a milk protein mixture useful for promoting growth of animal cells, for treating a surface wound, or for treating a gastrointestinal injury, disease, or ulcer, methods for preparing the milk protein mixture and methods employing the milk protein mixture. The milk protein mixture is prepared from a milk product such as cheese whey employing a cation exchange resin suitable for absorbing the milk protein mixture, filtering, and concentrating the product of cation exchange. The milk protein mixture for promoting growth of animal cells also includes a liquid culture medium. The milk protein mixture for treating a surface wound or for treating a gastrointestinal injury, disease, or ulcer also includes a pharmaceutically or veterinarily acceptable diluent, carrier, or excipient.

Abstract: Stem cell factor in combination with soluble interleukin-6 receptor, interleukin-6, for gp130 signaling, supports the proliferation, differentiation and terminal maturation of blood cells from normal human hematopoietic multipotential cells.

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 model utilizing cells for assessing oral bioavailability and potential drug-drug interactions of pharmacological agents is described. The model subjects cells (e.g., Caco-2 cells) to conditions that result in reliable expression of catalytically-active CYP3A4 at levels that appear to be comparable to levels present in mature enterocytes. These conditions include plating of selected clones on an extracellular matrix, exposure to 1.alpha.,25-(OH).sub.2 -D.sub.3 for a defined period of time, and the presence of serum in the medium. The model is useful for defining the role of CYP3A4 in limiting the oral bioavailability of many pharmacological agents and in drug-drug interactions involving CYP3A4 substrates that are believed to occur largely at the level of the intestine.

Abstract: A method for the in vitro proliferation and differentiation of neural stem cells and stem cell progeny comprising the steps of (a) isolating the cells from a mammal, (b) exposing the cells to a culture medium containing a growth factor, (c) inducing the cells to proliferate, and (d) inducing the cells to differentiate is provided.

Abstract: A method for producing proliferating cultures of dendritic cell precursors is provided. Also provided is a method for producing mature dendritic cells in culture from the proliferating dendritic cell precursors. The cultures of mature dendritic cells provide an effective means of producing novel T cell dependent antigens comprised of dendritic cell modified antigens or dendritic cells pulsed with antigen, including particulates, which antigen is processed and expressed on the antigen-activated dendritic cell. The novel antigens of the invention may be used as immunogens for vaccines or for the treatment of disease. These antigens may also be used to treat autoimmune diseases such as juvenile diabetes and multiple sclerosis.

Abstract: This invention provides a method for culturing dermal papilla cells with at least either of the mammalian epidermal cells from the sole or other portions of a mammal and the conditioned medium thereof, in order to permit long stable subculture of dermal papilla cells while keeping the original function thereof intact.

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: Methods for activating cytotoxic T lymphocytes (CTL) in vitro are presented in conjunction with methods for using the activated CTL for therapy in vivo. Additionally, a method for killing specific CTL in vivo is presented using antigen presenting cells which were modified in vitro.

Abstract: Novel media and methods are disclosed for the in vitro formation of a histologically complete human epithelium. The media are serum-free, companion cell or feeder layer free and organotypic, matrix free solutions for the isolation and cultivation of clonally competent basal epithelial cells. The media and methods of the invention are useful in the production of epithelial tissues such as epidermis, cornea, gingiva and ureter.

Abstract: The present invention relates to methods and compositions for eliciting an immune response and the prevention and treatment of primary and metastatic neoplastic diseases and infectious diseases. The methods of the invention comprise administering a composition comprising an effective amount of a complex, in which the complex consists essentially of a heat shock protein (hsp) noncovalently bound to an antigenic molecule in combination with administering antigen presenting cells sensitized with complexes of hsps noncovalently bound to an antigenic molecule. "Antigenic molecule" as used herein refers to the peptides with which the hsps are endogenously associated in vivo as well as exogenous antigens/immunogens (i.e., with which the hsps are not complexed in vivo) or antigenic/immunogenic fragments and derivatives thereof. In a preferred embodiment, the complex is autologous to the individual. In a specific embodiment, the effective amounts of the complex when administered intradermally are in the range of 0.

Abstract: All lines have been prepared from growth suppressor gene deficient animals. The cells include immortalized precursor cells and differentiated cells such as osteoclast precursors, osteoblast precursors, megakaryocytes, osteoclasts, osteoblasts, pancreatic .alpha.-cells, pancreatic .beta.-cells, pancreatic .delta.-cells, adipocytes, macrophages, chondrocytes, dendritic cells, hepatocytes, myocytes and prostatic cells. The cells are useful for constructing cDNA and protein libraries, screening agonists and antagonists of compounds and factors that affect metabolic pathways of specific cells and generating cell-specific antibodies.

Abstract: The present invention provides a rapid expansion method (termed "REM"), for quickly generating large numbers of T lymphocytes, including cytolytic and helper T lymphocytes. REM involves culturing the T cells in association with a disproportionately large concentration of nondividing feeder cells, preferably .gamma.-irradiated peripheral blood mononuclear cells ("PBMC") present at an excess of at least 40-fold (relative to the number of target T cells), more preferably at an excess of at least about 200-fold. Cultures grown under REM exhibit dramatically enhanced expansion rates that can be even further elevated by the use of appropriate concentrations of an additional feeder cell, an anti-CD3 monoclonal antibody and IL-2, as described herein. Clonal expansions in the range of 500-fold to 3000-fold can be achieved within a single stimulation cycle of about 10-13 days, which is more than 100-fold more efficient than currently employed methods of culturing human T cell clones.

Abstract: A method is disclosed for culturing animal cells in a medium characterized n that said medium contains a compound represented by the general formula ##STR1## wherein, R denotes COOM or CHO, herein M denotes hydrogen, an alkali metal or a C.sub.1 to C.sub.3 alkyl group, and n is an integer of 1 to 3.According to the present method, proliferation of animal cells can be enhanced with good reproducibility.

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: The invention provides a method for inhibition of proliferation of primitive hematopoietic progenitor cells and hematopoietic stem cells, by incubating preparations containing said cells with IFN-.gamma.. The method is especially useful for protecting said cells during purging techniques using cytotoxic treatment, and for protection of said cells during in vivo cytotoxic treatment.

Abstract: The present invention relates to a population of blood borne mammalian cells that express a unique profile of surface markers that includes certain markers typical of connective tissue fibroblasts, and are referred to herein as "blood-borne mesenchymal cells." In particular, it relates to the isolation, characterization and uses of such blood-borne mesenchymal cells. The cells of the present invention can be distinguished from peripheral blood leukocytes by their distinct size, morphology, cell surface phenotype and biologic activities, and are likewise distinguishable from connective tissue fibroblasts by other surface phenotypic markers. These cells proliferate in culture, and in vivo, as demonstrated in animal models, are capable of migrating into wound sites from the blood. Therefore, such blood-borne mesenchymal cells may have a wide range of applications, including, but not limited to, the promotion of wound healing, tissue remodeling, and for gene therapy.

Abstract: The present invention provides culture methods of T lineage precursor cells comprising a step of culturing the T lineage precursor cells under a condition that a dissolved oxygen concentration in the nutrient medium is higher than a dissolved oxygen concentration in the nutrient medium when the medium is in contact with normal atmospheric air. According to the methods of the present invention, by using a submerged (suspension) culture which is a general method for culturing cells or tissues, the derivation of matured T cell(s) differentiated from T precursor cells in terms of their phenotypes of differentiation antigens and of their functions is realized to be accomplished.

Abstract: Provided are methods and compositions for the repair of articular cartilage defects in a mammal. Denuded chondrogenic cells are proliferated ex vivo as monolayer cultures in order to expand the pool of available chondrogenic cells. During proliferation the chondrogenic cells stop secreting the extracellular matrix components, type II collagen and sulfated proteoglycans. The proliferated cells then are seeded into a pre-shaped well having a cell contacting, cell abhesive surface. The cells cultured in the well redifferentiate and begin to secrete cartilage-specific extracellular matrix again. Accordingly, essentially unlimited amounts of synthetic cartilage may be prepared from small samples of biopsy tissue. Also provided are methods for surgically repairing articular cartilage defects in mammals using the synthetic cartilage prepared in accordance with the invention.