Abstract: The present invention fulfills a need in the art for methods that promote hematopoietic and mesenchymal stem and lineage-specific cell proliferation and differentiation by growth in the presence of angiotensinogen, angiotensin I (AI), AI analogues, AI fragments and analogues thereof, angiotensin II (AII), AII analogues, AII fragments and analogues thereof and AII AT2 type 2 receptor agonists.

Abstract: Bone marrow stromal cells (BMSC) differentiate into neuron-like phenotypes in vitro and in vivo, engrafted into normal or denervated rat striatum. The BMSC did not remain localized to the site of the graft, but migrated throughout the brain and integrated into specific brain regions in various architectonic patterns. The most orderly integration of BMSC was in the laminar distribution of cerebellar Purkinje cells, where the BMSC-derived cells took on the Purkinje phenotype. The BMSC exhibited site-dependent differentiation and expressed several neuronal markers including neuron-specific nuclear protein, tyrosine hydroxylase and calbindin. BMSC can be used to target specific brain nuclei in strategies of neural repair and gene therapy.

Abstract: The present invention provides methods for generating antigen-reactive T cells in vitro comprising priming immune cells and incubating the primed immune cells in vitro with a non-covalent complex of an heat shock protein and an antigenic molecule. The present invention further relates to methods for generating antigen-reactive CD4+ T cells for immunotherapy. Methods and compositions are also disclosed for the treatment and prevention of cancer or infectious disease in a subject comprising administering to the subject MHC matched antigen-reactive T cells that are generated in vitro by the present methods.

Abstract: Treatment of tumors, including their metastases, is described. Retrieved cytokines and other molecules from the growth medium of human monocytes stimulated ex vivo with gamma globulin, or other immune stimulators are employed for cancer therapy.

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: A peptide containing a sequence for the first fifteen amino acids from the N-terminal of Asn-Leu-Val-Glu-Phe-Gly-Lys-Met-Ile-Glu-Cys-Ala-Ile-Arg-Asn is used in a cell culture medium to promote cell growth in vitro and in the treatment of wounds. A method of preparation of the peptide is also claimed.

Abstract: Disclosed are optimized methodologies for isolating and propagating stem cells from biopsies and postmortem tissues. Specifically disclosed are methods of culturing neural stem cells in the presence of a cocktail of trophic factors/co-factors for enhanced propagation.

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: This disclosure provides an improved system for culturing human pluripotent stem (pPS) cells in the absence of feeder cells. The role of the feeder cells can be replaced by supporting the culture on an extracellular matrix, and culturing the cells in a conditioned medium. Permanent cell lines are provided that can produce conditioned medium on a commercial scale. Methods have also been discovered to genetically alter pPS cells by introducing the cells with a viral vector or DNA/lipid complex. The system described in this disclosure allows for bulk proliferation of pPS cells for use in studying the biology of pPS cell differentiation, and the production of important products for use in human therapy.

Abstract: This invention relates to the discovery that an intermediate, differentiated stage of pancreatic stem cells exist that can be propagated in a stable manner in successive serial passaging while maintaining insulin production in response to glucose. These cells are advantageous in that they are both expandable and stable in culture and can driven to late stage development, i.e. prototype islet cells. This invention further provides for culturing techniques that select for these intermediate differentiated stage cells and selectively eliminates early or late stage pancreatic cells.

Abstract: The present invention relates generally to growth factors and more particularly to growth factors which are capable of stimulating or otherwise facilitating formation of insulin-secreting cells. The identification of these growth factors permits the development of protocols to culture cells in vitro for transplantation into mammalian and in particular human subjects with insulin-dependent type 1 diabetes or related conditions. It is further contemplated that the endogenous expression of growth factors required for the development of insulin-producing cells may be manipulated in vivo, by the appropriate administration of agents including genetic agents capable of regulating the expression of growth factors in pancreatic duct epithelial cells. The growth factors ray also be administered to subjects with type 1 diabetes to stimulate the proliferation and differentiation of pancreatic cells into insulin-secreting cells.

Abstract: The present invention provides a process of biosynthesizing papillomavirus by inducing complete differentiation of an epithelial cell that contains papillomavirus DNA. Complete differentiation is induced by exposing epithelial cells to a protein kinase C inducer. Assays for screening agents that modify papillomavirus biosynthesis, determining the papillomavirus infectivity of epithelial cells, detecting the presence of anti-papillomavirus antibodies and vaccinating against papillomavirus infection are also provided.

Abstract: The present invention relates generally to nutritive medium, medium supplement, media subgroup and buffer formulations. Specifically, the present invention provides powder nutritive medium, medium supplement and medium subgroup formulations, particularly cell culture medium supplements (including powdered sera such as powdered fetal bovine serum (FBS)), medium subgroup formulations and cell culture media comprising all of the necessary nutritive factors that facilitate the in vitro cultivation of cells. The invention further provides powder buffer formulations that produce particular ionic and pH conditions upon reconstitution with a solvent.

Abstract: Disclosed are compositions and methods for accelerating the differentiation of human MSCs into the osteogenic lineage with resultant enhanced de novo bone formation. This makes possible improved methods for bone defect repair. Thus, one aspect of the invention is a method for accelerating the differentiation of ALCAM-bearing hMSCs into the osteogenic lineage by contacting such hMSCs with a ligand that binds to activated leukocyte-cell adhesion molecule (ALCAM) on a single hMSC. Preferably, the ligand is a Fab fragment of a monoclonal antibody expressed by the hybridoma of ATCC Accession No. NB 11789.

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: Disclosed is a process for preparing agents containing virus-inactivated vitamin K-dependent plasma components as well as protein C, protein S, factors II, VII, IX and/or X as well as combinations thereof, such as, for example, PPSB preparations, wherein a source containing these components is subjected to appropriate separation procedures, especially by using membrane-chromatographic methods.

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: A serum-free medium which supports the proliferation and differentiation of CD34+ 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+ cells for use in human therapeutic protocols. The cells CD34+ cultured in a the medium of the present invention are used in a method for repopulating human host bone marrow.

Abstract: A process is provided for expanding the population of endothelial cells obtained from peripheral blood which can be transformed with a vector comprising a DNA sequence encoding a preselected bioactive polypeptide. The resulting transgenic endothelial cells are useful to biocompatibilize implantable medical devices or can be used directly, as for gene therapy.

Abstract: Disclosed are methods for the isolation of primordial germ cells, culturing these cells to produce primordial germ cell-derived cell lines, methods for transforming both the primordial germ cells and the cultured cell lines, and using these transformed cells and cell lines to generate transgenic animals. The efficiency at which transgenic animals are generated by the present invention is greatly increased, thereby allowing the use of homologous recombination in producing transgenic non-rodent animal species.

Abstract: Endothelial cells have the natural property of releasing soluble factors into the fluid surrounding them, said factors altering the behavior of immune cells. In cell systems containing at least endothelial cells, the release of this type of factor, which promotes the proliferation of resting and weakly activated lymphocytes and at the same time inhibits the proliferation of highly activated lymphocytes and transformed lymphoblasts without impairing their other vital functions, is induced by administration of pentacyclic oxindole alkaloids but inhibited by the concurrent administration of tetracyclic oxindole alkaloids.

Abstract: Cells, preferably isolated pancreatic islet cells, are treated with a compound or a combination of compounds selected from an antioxidant, an anti-cytokine, an anti-endotoxin and an antibiotic. The compound or combination of compounds is in a medium for culturing the cells before microencapsulation, in a medium for cryopreserving the cells by freezing followed by thawing and microencapsulation, in a medium for culturing the cells after microencapsulation, or in a medium for culturing the cells before microencapsulation and in a medium for culturing the cells after microencapsulation. Microcapsules containing cells may be incubated with a physiologically acceptable salt to increase durability of the microcapsules. In a preferred method, isolated pancreatic islet cells are cultured for about 12 to about 36 hours in a medium containing an antioxidant as the compound, cryopreserved by freezing in a medium containing the compound or combination of compounds, thawed and microencapsulated.

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.

Abstract: A method for obtaining lineage committed human cells imbued with enhanced proliferative potential, biological function, or both, comprising culturing lineage committed human cells under physiologically acceptable liquid culture conditions, where the liquid culture medium is replaced at a rate and for a time sufficient to obtain the human lineage committed cells imbued with enhanced proliferative potential, biological function, or both; and isolating the cultured cells.

Abstract: The present invention relates to methods for using various forms of a novel receptor expressed by hematopoietic and endothelial cells. An additional variant form of this receptor has been detected in brain cells and shown to bind to the obese gene product, leptin. Therefore, leptin may be used to stimulate the growth and development of receptor-positive hematopoietic and endothelial cells in vitro and in vivo. In addition, this receptor is selectively expressed in hematopoietic progenitor cells with long-term repopulating potential. Thus, agents that specifically bind to this receptor may be used to identify and isolate progenitor cells for a variety of clinical applications.

Abstract: Methods, compositions and devices are provided for the growth of hematopoietic cells in culture. Bioreactors are provided in which diverse cell types are simultaneously cultured in the presence of appropriate levels of nutrients and growth factors substantially continuously maintained in the bioreactor while removing undesirable metabolic products. This simultaneous culture of multiple cell types is required for the successful reconstruction of hematopoietic tissue ex vivo. At least one growth factor is provided through excretion by transfected stromal cells, particularly heterologous cells. Means are provided for maintaining the stromal cells and hematopoietic cells separately, to allow for early removal of the hematopoietic cells.

Abstract: A method of pretreating healthy donor's myoblast cultures with growth or trophic factors like basic fibroblast growth factor (bFGF) and with concanavalin A on transplantation to subjects suffering of myopathy like muscular dystrophy is disclosed and claimed. Recipient muscles show a higher percentage of functional cells, a four-fold increase, demonstrated by the higher incidence of dystrophin-positive fibers, and does not require previous preconditioning of recipient muscles by irradiation or toxin administration. The recipient subjects were immunosuppressed with FK 506. When growing myoblasts with 20 &mgr;g/ml concanavalin A or 100 ng/ml TPA for two to four days, migration of donor cells in recipient tissue was increased by 3-4 fold.

Abstract: Methods for clearing Chlamydia from biological materials, e.g., cells and animals, infected therewith are described. Methods for maintaining Chlamydia-free cells and animals are also described.

Abstract: The present invention relates generally to nutritive medium, medium supplement, media subgroup and buffer formulations. Specifically, the present invention provides powder nutritive medium, medium supplement and medium subgroup formulations, particularly cell culture medium supplements (including powdered sera such as powdered fetal bovine serum (FBS)), medium subgroup formulations and cell culture media comprising all of the necessary nutritive factors that facilitate the in vitro cultivation of cells. The invention further provides powder buffer formulations that produce particular ionic and pH conditions upon reconstitution with a solvent.

Abstract: Growth of keratinocytes is provided by subjecting a basal medium, insulin, pituitary compound and keratinocytes to cell growth conditions. Cell growth can be provided in the absence of exogenous epidermal growth factor. A conditioned medium results after the keratinocytes are subjected to cell growth conditions, treated keratinocytes are provided, and the treated keratinocytes are subjected to cell growth conditions in a basal medium. The conditioned medium contains autocrine factor. The conditioned medium then is contacted with further keratinocytes to provide a cell growth mixture. The cell growth mixture is subjected to incubation in order to provide replicative DNA synthesis and hence cell proliferation. Cells so provided can be used for dermatological research, toxicological testing and skin grafts.

Abstract: The present invention relates to methods and compositions for eliciting an immune response and the prevention and treatment of primary and metastatic noeplastic 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.

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. The compounds further delay senescence, optimize growth and maturation, and increase population doublings.

Abstract: The present invention provides a modified rapid expansion method (termed “low-PBMC-REM” or “modified-REM”), for quickly generating large numbers of T lymphocytes, including cytolytic and helper T lymphocytes, without using the large excesses of peripheral blood mononuclear cells (PBMC) or EBV-transformed lymphoblastoid cells (LCL) characteristic of high-PBMC-REM. Clonal expansions of greater than 500-fold can be achieved within a single stimulation cycle of about 8-14 days.

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 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: Cells, preferably isolated pancreatic islet cells, are treated with a compound or a combination of compounds selected from an antioxidant, an anti-cytokine, an anti-endotoxin and an antibiotic. The compound or combination of compounds is in a medium for culturing the cells before microencapsulation, in a medium for cryopreserving the cells by freezing followed by thawing and microencapsulation, in a medium for culturing the cells after microencapsulation, or in a medium for culturing the cells before microencapsulation and in a medium for culturing the cells after microencapsulation. In a preferred method, isolated pancreatic islet cells are cultured for about 12 to about 36 hours in a medium containing an antioxidant as the compound, cryopreserved by freezing in a medium containing the compound or combination of compounds, thawed and microencapsulated. The microcapsule contains a hydrogel core such gelled alginate and a semipermeable outer membrane such as formed with polylysine.

Abstract: A novel growth factor, artemin, which belongs to the GDNF/neurturin/persephin family of growth factors, is disclosed. The human and mouse amino sequences have been identified. Human and mouse artemin genomic DNA sequences have been cloned and sequenced and the respective cDNA sequences identified. In addition, methods for treating degenerative conditions using artemin, methods for detecting artemin gene alterations and methods for detecting and monitoring patient levels of artemin are provided.

Abstract: Cell compositions from which cancer cells have been selectively eliminated by using apoptosis inducers specific to cancer cells.

Abstract: Disclosed are methods for the isolation of primordial germ cells, culturing these cells to produce primordial germ cell-derived cell lines, methods for transforming both the primordial germ cells and the cultured cell lines, and using these transformed cells and cell lines to generate transgenic animals. The efficiency at which transgenic animals are generated by the present invention is greatly increased, thereby allowing the use of homologous recombination in producing transgenic non-rodent animal species.

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: A method is described whereby dendritic cells derived from the CD34+ and CD34− 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: 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: &bgr;-alethine is employed in the differentiation, phenotypic expression, and vitalization of cells, for both in vivo and in vitro applications. Particular applications include the use of &bgr;-alethine in the treatment of immune disorders and diseases, and in the promotion of cell cultures.

Abstract: Primordial germ cells isolated from human embryonic tissue, such as from the gonadal ridges of human embryo, are disclosed. The primordial germ cells are cultured resulting in cells that resemble embryonic stem cells or embryonic germ cells in morphology and pluripotency. The cells are maintained several months in culture and can be genetically manipulated using transgenic technology to insert heterologous genetic material.

Abstract: A serum-free medium which supports the proliferation and differentiation of CD34+ 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+ cells for use in human therapeutic protocols. The cells CD34+ cultured in a the medium of the present invention are used in a method for repopulating human host bone marrow.

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: The invention relates, inter alia, to a process for in vitro culture of any cells whatsoever, of human or animal origin. The process's characterizing feature is that it involves at least a stage of transitional incubation of the said cells, isolated in an incubation medium in the form of a solution whose constituents are chosen in such a manner that the final calcic concentration of the solution is between [0-1 mM], and preferably [0-100 &mgr;M], to induce rapid and massive adherence of the isolated cells to any given support in contact with the said incubation medium. Applications to gastrointestinal epithelium cell culture and vaccine production in fermenter units.

Abstract: A method of treatment using immune system suppressor cells and immune system stimulator cells comprises providing stem cells; combining the stem cells with lymphoid-derived cells to produce a co-culture; adding lipopolysaccharide and a factor selected from the group consisting of granulocyte macrophage-colony stimulating factor, macrophage colony stimulating factor, granulocyte-colony stimulating factor and mixtures thereof to the co-culture; obtaining immune system suppressor cells and immune system stimulator cells from the co-culture; and introducing and the cells into a host.

Abstract: One object of the present invention is based upon the development and use of a serum-free defined cell culture medium comprising a supplement mixture, a component mixture, a vitamin mixture, an inorganic salt mixture and amino acid mixture that avoids the problems inherent in the use of serum. In particular, the defined medium is useful in culturing fibroblasts, especially chondrocytes. Another object of the present invention is to claim a method of enhancing the differentiation of chondrocytes and enhancing the synthesis of a cartilage specific matrix using tumor growth factor beta (TGF-.beta.). Another object of the present invention is to claim a method of enhancing the differentiation of chondrocytes using the combination of TGF-.beta.and IGF.

Abstract: The present invention provides a method for cultivation of hepatocytes efficiently by using a hepatocyte growth promoting factor derived from 3T3 cells, which comprises culturing hepatocytes isolated from the liver of a matured mammal in a medium containing pleiotrophin, fetal bovine serum, ascorbic acid or analogues thereof and nicotinamide or an analogue thereof, thereby forming primary colonies of the hepatocytes; and a method for subculture of the primary hepatocytes in the same medium.