Abstract: The present invention relates to a growth factor preparation derived from a mixed lymphocyte culture, and a process for its production. The invention also relates to a cell culture medium containing said growth factor preparation. The invention further relates to a process for culturing plasma cells and producing antibodies by using said cell culture medium.

Abstract: A procedure for carrying out T cell differentiation of CD34+ stem cells in an in vitro culture of thymic epithelial fragments whereby the differentiated T cells achieve full immunocompetence. The invention also includes the procedure for differentiation of stem cells from HIV seropositive individuals or genetically modified stem cells. The invention broadly relates to the culture of cultured thymic epithelial fragments and provides procedures for verifying true immunocompetence of the resulting T cells and for analyzing the effects of various compounds on the differentiation process. The invention also comprises several novel applications for utilizing the procedure of the invention, including grafting fortified cultured thymic epithelial fragments and infusing immunocompetent T cells into patients with compromised immune systems.

Abstract: A method for producing genetically modified neural cells comprises culturing cells derived from embryonic, juvenile, or adult mammalian neural tissue with one or more growth factors that induce multipotent neural stem cells to proliferate and produce multipotent neural stem cell progeny which include more daughter multipotent neural stem cells and undifferentiated progeny that are capable of differentiating into neurons, astrocytes, and oligodendrocytes. The proliferating neural cells can be transfected with exogenous DNA to produce genetically modified neural stem cell progeny. The genetic modification can be for the production of biologically useful proteins such as growth factor products, growth factor receptors, neurotransmitters, neurotransmitter receptors, neuropeptides and neurotransmitter synthesizing genes.

Abstract: The use of individual or combinations of cytokines, particularly IL-3, GM-CSF, and c-kit ligand are employed for long-term hematopoiesis in serum free culture in the absence of stromal cells. The cultures can be used for evaluating compounds and their effect on hematopoiesis, particularly as to lifetime and nature of differentiation. In addition, the expanded cells may be used for engraftment in a mammalian host or enhancement of particular cell lineages in a mammalian host. The subject systems may be used with any mammalian hemopoietic cells, but finds particular application with primates, more particularly humans.

Abstract: The present invention provides methods and bioreactors for expanding stem cells in a population of cells substantially enriched in hematopoietic stem cells and substantially free of stromal cells. The method comprising the steps of inoculating the population of cells in an expansion container in a volume of suitable medium such that the cell density is at least about 5,000 cells/1 mL and at an initial oxygen concentration of less than 8%; adding an effective amount of at least one cytokine to cause stem cell expansion; culturing the cells under suitable conditions such that the cells condition the medium; increasing the oxygen concentration to about 20%; exchanging the medium at a rate which allows expansion of the stem cells; and culturing the cells under conditions such that the stem cells are expanded. The present invention also provides a bioreactor constructed to accomodate the operational requirements for stroma-free stem cell expansion.

Abstract: Non-carboxylated sulfated polyanions have been successfully used to rapidly obtain and maintain stable single-cell suspension of BTI-TN5B1-4 cells, a cell line which has a high intrinsic capacity for the expression of recombinant protein, but which clumps severely in suspension reducing its effectiveness as a host for foreign protein production with the baculovirus expression vector system. The three most effective polyanions for inducing a single-cell suspension were dextran sulfate, polyvinyl sulfate, and pentosan sulfate. The cost of dextran sulfate treatment is low compared to heparin treatment, which required a 20-fold higher lever to induce single-cell suspension. More importantly, dextran sulfate does not block vital infection at MOI.gtoreq.1 whereas heparin is known to seriously inhibit infection.

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: The present invention provides systems, methods and chemically defined media for the cultivation of cells, particularly epithelial cells. Cells may be cultured with a varied calcium concentration. Furthermore, a calcium concentation in excess of 1.00 mM may be used in the practice of the present invention without loss of a proliferative cell population and maintianing high colony forming efficiencies. Population doubling times range from about 16 to about 33 hours. Cells may serially cultivated to achieve from about 20 to about 50 population doublings.

Abstract: Immortal avian T lymphocyte cell lines which produce and secrete immune lymphokines are disclosed. These cell lines may be produced from T cells recovered from fowl which have been hyperimmunized in vivo. The activated T cells are first exposed in vitro to a mitogen effective for secondary stimulation thereof, and then virally transformed to produce an immortal cell line. When cultured in vitro, the cell lines produce and secrete immune lymphokines which may be administered to fowl to increase their resistance to infections.

Abstract: Methods and formulations are disclosed for the in vitro formation of a histologically complete human epidermis in a serum-free, companion cell or cell feeder layer-free, and organotypic matrix-free culture system commencing with the isolation and cultivation of a unique population of clonally-competent basal epidermal cells and ending with the formation of a functional, histologically complete, human squamous epithelium. The formation of a histologically complete human epidermis is accomplished in a serum-free medium, without companion-cells or feeder layer cells or any organotypic support using a multi-step process that is controlled by manipulating the growth and differentiation factors requisite to the sequential development of a usable, functional, and completely differentiated epidermis.

Abstract: Provided are serum-free media for tissue culture containing tissue inhibitor of metalloproteinases and methods for culturing animal cells using these media.

Abstract: Methods, including culture media conditions, which provide for ex vivo 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 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: This invention is directed to a method for stimulating the proliferation of V.gamma.2V.delta.2 T cells comprising contacting V.gamma.2V.delta.2 T cells with a V.gamma.2V.delta.2 T cell proliferation stimulating amount of a compound selected from the group consisting of a monoalkyl phosphate and an alkenyl pyrophosphate.

Abstract: Methods for increasing the number of human hematopoietic precursor cells in vitro are provided. The methods generally comprise (a) separating human hematopoietic precursor cells from mature hematopoietic cells present in a blood product; (b) inoculating the separated precursor cells into a culture vessel containing a culture medium comprising a nutritive medium and a source of growth factors at a density of between 1.times.10.sup.3 cells/ml and 4.times.10.sup.6 cells/ml; and (c) culturing the cells under conditions and for a time sufficient to increase the number of precursor cells relative to the number of such cells present in the blood product. The culture medium may also include a suitable amount of microcarrier beads. Suitable blood products include bone marrow, umbilical cord blood, and peripheral blood. A device for carrying out such methods is also provided.

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: Fibroblast growth factors are used in vivo, in situ and in vitro to stimulate stem cells, hemopoiesis, the immune system, transplant donor cells, culture and/or engraftment, wherein the use of fibroblast growth factors is disclosed for the stimulation of stem cells or hemopoietic cells, supporting cells and their progeny, in vitro, in situ and in vivo, as well as corresponding engrafting sites in vivo.

Abstract: Stem cell factor in combination with interleukin-6 and soluble interleukin-6 receptor supports proliferation, differentiation and terminal maturation of erythroid cells from normal human hematopoietic stem cells.

Abstract: In vitro production of clinically useful quantities of mature, differentiated human blood cells 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 recombinant human growth and maturation promoting polypeptide factors in combinations that expand stem cell cultures and promote the maturation and differention of stem cells into 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 modified so as to remove histocompatibility and/or blood group antigens, or may be genetically altered by transfection with appropriate DNA-containing vectors, prior to addition to the bioreactor.