Abstract: The present invention relates to methods of constructing an integrated artificial immune system that comprises appropriate in vitro cellular and tissue constructs or their equivalents to mimic the normal tissues that interact with vaccines in mammals. The artificial immune system can be used to test the efficacy of vaccine candidates in vitro and thus, is useful to accelerate vaccine development and testing drug and chemical interactions with the immune system.

Abstract: Devices, in vitro cell cultures, systems, and methods are provided for microscale cell culture analogous (CCA) device.

Abstract: Disclosed herein is a system and method for producing T cells from stem cell populations. Specifically exemplified herein is a culture system and method that produces CD4 cells and/or T cell subtypes from a CD4 lineage using a sample of hematopoietic stem cells. Adult hematopoietic precursor/stem cells (HPC) are progenitors to all lineages of immune cells. There has been limited success in generating functional CD4 T cells with this convenient culture system. Also disclosed herein is a novel stromal cell line expressing DL1, interleukin-7 (IL-7), and FMS-like tyrosine kinase 3 ligand (Flt3-L). This improved culture system can greatly facilitate the study of late T cell development and enables immunotherapeutic applications.

Abstract: Provided are an iPS cell derived from a somatic cell such as an NKT cell, having the ?-chain region of the T cell antigen receptor gene rearranged to uniform V?-J? in an NKT cell receptor-specific way, NKT cells differentiated from the iPS cell, a method of creating the same, and an immune cell therapy agent prepared using cells differentiated from the iPS cell.

Abstract: A method of producing a restorative material used to restore a tooth-deficient area in an oral cavity, the method comprising: positioning, in a support carrier, a first cell mass formed from either mesenchymal cells or epithelial cells and a second cell mass formed from the other of the mesenchymal cells or epithelial cells, one of the mesenchymal cells or epithelial cells being derived from a tooth germ and the first and second cell masses being not mixed with each other but made to closely contact each other; culturing the first and second cell masses to form a reconstructed tooth germ or tooth; and confirming directionality of the reconstructed tooth germ or tooth formed by the culturing so as to enable the reconstructed tooth germ or tooth to be embedded in the tooth-deficient area such that a tip of the tooth faces an interior of the oral cavity, the tooth germ or tooth whose directionality has been confirmed being used as a restorative material to obtain an equivalent of a missing tooth in the tooth-def

Abstract: The present invention incorporates germinal centers (GCs) into three-dimensional (3D) engineered tissue constructs (ETCs). In an embodiment, we have incorporated the GC in the design of an artificial immune system (AIS) to examine immune responses to vaccines and other compounds. Development of an in vitro GC adds functionality to an AIS, in that it enables generation of an in vitro human humoral response by human B lymphocytes that is accurate and reproducible, without using human subjects. The invention also permits evaluation of, for example, vaccines, allergens, and immunogens, and activation of human B cells specific for a given antigen, which can then be used to generate human antibodies. In an embodiment of the present invention the function of the in vitro GC is enhanced by placing FDCs and other immune cells in a 3D ETC; FDCs appear more effective over a longer time (antibody production is sustained for up to about 14 days.

Abstract: Methods for the diagnosis of visceral, cutaneous and canine leishmaniasis in a subject suspected of being infected with the parasitic protozoa Leishmania is disclosed. Disclosed are antibody-capture enzyme-linked immunosorbent assays (ELISAs) for the detection of antibodies to Leishmania parasite soluble antigens and antigen-capture ELISAs for the detection of Leishmania parasite soluble antigens in host samples. Also disclosed are immunodiagnostic kits for the detection of Leishmania parasite circulating antigens or IgM and IgG antibodies in a sample from subject having visceral, cutaneous or canine leishmaniasis. In these methods and kits, detection may be done photometrically or visually. The methods and kits also allow the visualization of Leishmania amastigotes or promastigotes in a sample.

Abstract: Novel antigen-presenting cells, including but not limited to dendritic cells, that are loaded with antigens from dead or dying cells including allogenic cell lines, and the methods for making such antigen-presenting cells are described. These loaded antigen-presenting cells induce therapeutic immune responses in humans. Such loaded antigen-presenting cells are useful in the management of cancer. Antigen-loaded dendritic cells prepared as described here can prime naïve T cells to differentiate into effector cells able to recognize multiple and/or shared tumor antigens that are expressed either on the tumor cells that are used to load the dendritic cells and/or on other tumor cells. The cytotoxic T cells generated by exposure to antigen-loaded dendritic cells prepared as described here can be used in adoptive therapy. This induction of responses against multiple antigens shared between different cells, for instance tumor cells, as described here is important as it leads to broad immune responses.

Abstract: Polynucleotides associated with differentiation states of stem cells are provided. Also provided are methods and kits for detecting, identifying and/or discriminating differentiated stem cells from undifferentiated ones by measuring an expression level of one or more genes, such as CBARA1 and LHX 6, in the stem cells.

Abstract: The present invention discloses novel dendritic cell maturation-inducing cytokine cocktails, and methods for inducting type-1 polarized dendritic cells in serum-free conditions which enhance the desirable properties of DC1s generated in serum-supplemented cultures. The invention further discloses methods and systems using IFN? and other ligands of the IFN?receptor, in combination with IFN? (or other type I interferons), poly I:C, and other IFN? (and IFN?) inducers to enhance the IL-12-producing properties of dendritic cells. More specifically, the present invention discloses type-1 polarized dendritic cells that have a unique combination of a fully-mature status and an elevated, instead of “exhausted”, ability to produce IL-12p70 allows for the generation of fully-mature DC1s in serum-free AIM-V medium.

Abstract: Purified totipotent stem cells and pluripotent stems cells derived by somatic cell nuclear transfer are disclosed herein, as well as cell lines, multipotent cells and differentiated cells produced from these stem cells. The stem cells are produced from an enucleated host cell from a first donor and nuclear genetic material from a somatic cell of a second donor. Methods for making and using such compositions of such stem cells are also provided.

Abstract: The present invention provides a screening method for somatic cell nuclear reprogramming substances, which comprises (a) a step for bringing into contact with each other a somatic cell comprising a gene wherein a marker gene is present at a position permitting expression control by the expression control region of an ECAT gene, and a test substance, and (b) a step following the aforementioned step (a), for determining the presence or absence of the emergence of cells expressing the marker gene, and selecting a test substance allowing the emergence of the cells as a somatic cell nuclear reprogramming substance candidate, and the like.

Abstract: Strains of Lactobacillus which have been selected for their capability of reducing gastrointestinal inflammation, such as that due to Helicobacter pylori, and products derived from these strains, including agents for treatment or prophylaxis of inflammation associated with Helicobacter pylori for administration to humans and include conditioned media in which the selected strains have grown and protein-containing extracts of the conditioned media.

Abstract: It is an object of the present invention to provide an immunodeficient animal capable of generating human-derived lymphoid cells, a human-derived lymphoid cell, and a method for producing a human antigen-specific antibody. The means for solving the aforementioned object is: an immature immunodeficient mammal into which human-derived hematopoietic precursor cells have been transplanted, and which is able to generate said human-derived hematopoietic cells or immunocompetent cells; and a method for producing a human-derived antibody, which is characterized in that it comprises recovering immunocompetent cells from the above-described mammal, culturing the immunocompetent cells, and collecting a human-derived antibody from the obtained culture product.

Abstract: An improved method of culturing cells for cell therapy applications that includes growing desired cells in the presence of antigen-presenting cells and/or feeder cells and with medium volume to surface area ratio of up to 1 ml/cm2 if the growth surface is not comprised of gas permeable material and up to 2 ml/cm2 if the growth surface is comprised of gas permeable material. The desired cells are at a surface density of less than 0.5×106 cells/cm2 at the onset of a production cycle, and the surface density of the desired cells plus the surface density of the antigen presenting cells and/or feeder cells are at least about 1.25×105 cells/cm2.

Abstract: Methods for obtaining pluripotent (embryonic stem) cells from parthenogenetic embryos, especially primates, are provided. These cells are useful for producing differentiated cells, tissues and organs, especially human and non-human primate cells, tissues and organs.

Abstract: This invention provides a co-culture method using 3-dimensional feeders to support single cell in microwells of microarray chips. Microbeads are utilized as carrier to manipulate feeders into 3-dimensional layers in a microwell, to retain feeders at desired location, to keep feeders away from single cell at a desired distance, to revive feeders for optimized co-culture, and to eliminate feeders from image background in post imaging analysis.

Abstract: A method is disclosed for making an artificial micro-gland having a continuous anisotropic membrane of two or more types of living cells. A first step includes forming a carrier fluid in a microchannel in a laminar flow of two distinct fluid flows. Another step includes introducing a template, which may itself be anisotropic, into the microchannel in a manner such that the template straddles the interface between the first fluid-flow and the second fluid-flow. In some embodiments two types of living cells within the template are separately attracted one of the fluid flows by the presence of an agent of taxis. In other embodiments, cells within one or the other of the fluid flows are attracted to agents within the template. Membranes form on the template and join together to form a complete cellular membrane around a reservoir.

Abstract: A perfusable bioreactor is used for the production of human tissues, animal tissues or tissue equivalents. Their production is based on a structure cultivated in the interior, the interior being enclosed by an envelope and containing at least one inlet and one outlet for a liquid nutrient medium. Accordingly, the bioreactor can be connected to a unit for producing a perfusion pressure of the nutrient medium. This tissue replacement is particularly suitable for use in clinical/therapeutical applications.

Abstract: A well-based flow system for cell culture is described which provides for flow of culture containing compounds for drug screening to be exposed to cells seeded on a membrane. The flow of medium may be planar or radial and means are provided for the removal of waste media through fluid outlets in fluid communication with the assay well plates through conduits. Methods of using the system for cell culture and drug toxicity screening are also provided including coculturing cells such as hepatocytes, stem cells, fibroblasts and smooth muscle cells and selectively exposing cells to test compounds.

Abstract: The present invention relates to a method for producing hair microfollicles comprising the steps of: a) providing de novopapillae, b) providing other cell populations selected from the group of fibroblasts, keratinocytes and melanocytes, and co-culturing the de novopapillae with at least one other cell population in non-adherent culture vessels. The present invention relates also to methods of producing de novo papillae usable in said method for producing hair microfollicles.

Abstract: Microfluidic bioreactor devices may feature intercommunicating microchannels defined in two polymer layers separated by a membrane. A geometric parameter associated with the microchannels in one layer may vary along a length of those channels.

Abstract: Biocompatible synthetic or natural scaffolds are provided for the reconstruction, repair, augmentation or replacement of organs or tissue structures in a patient in need of such treatment. The scaffolds are shaped to conform to at least a part of the organ or tissue structure and may be seeded with one or more cell populations. Inserts, receptacles and ports are also provided for the attachment of tubular vessels to the neo-organ scaffolds. The seeded scaffolds are implanted into the patient at the site in need of treatment to form an organized organ or tissue structure. The scaffolds may be used to form organs or tissues, such as bladders, urethras, valves, and blood vessels.

Abstract: The invention provides means and methods for providing a cell with a spontaneous electrical activity and means and methods for increasing the depolarization rate of a cell having a spontaneous electrical activity. Means and methods are provided comprising: providing a cell with a compound capable of providing and/or increasing a pacemaker current If, and diminishing electrical coupling between said cell and surrounding cells and/or reducing the inward rectifier current IK1 of said cell, and or increasing the availability of INa at depolarized potentials, preferably using overexpression of additional sodium channels.

Abstract: The present invention provides methods for the culture of animal pluripotent stem cells and their differentiated progeny cells, tissues, and organs, and nonhuman animal embryos and fetuses.

Abstract: The present invention is a method for inducing cytotoxic T cell having an antigen-specific cytotoxic activity, a method for maintaining the cell, a method for continuously culturing the cell or a method for expanding the cell, comprising the step of culturing a cytotoxic T cell in the presence of at least one substance selected from the group consisting of (A) a substance having a binding activity to CD44; (B) a substance capable of regulating a signal emitted by binding a CD44 ligand to CD44; (C) a substance capable of inhibiting binding of a growth factor to a growth factor receptor; (D) a substance capable of regulating a signal emitted by binding of a growth factor to a growth factor receptor; and (E) fibronectin, a fragment thereof or a mixture thereof.

Abstract: This invention provides a system for obtaining cells of the chondrocyte lineage by differentiating primate pluripotent stem cells. The process involves culturing the cells as a micromass or other aggregate form in a cocktail of differentiation agents that facilitates outgrowth of the desired cell type. Progeny are capable of synthesizing Type II collagen or aggrecan, or other products that are characteristic of the chondrocyte lineage. Chondrocytes and chondrocyte precursor cells obtained according to this disclosure are suitable for use in both research and clinical therapy.

Abstract: Methods and devices for applying hemodynamic patterns to human/animal cells in culture are described. Hemodynamic flow patterns are measured directly from the human circulation and translated to a motor that controls the rotation of a cone. The cone is submerged in fluid (i.e., cell culture media) and brought into close proximity to the cells. Rotation of the cone creates time-varying shear stresses. This model closely mimics the physiological hemodynamic forces imparted on endothelial cells in vivo. A TRANSWELL coculture dish (i.e., a coculture dish comprising an artificial porous membrane) may be incorporated, permitting two, three, or more different cell types to be physically separated within the culture dish environment. In-flow and out-flow tubing may be used to supply media, drugs, etc. separately and independently to both the inner and outer chambers. The physical separation of the cell types permits each cell type to be separately isolated for analysis.

Abstract: The present invention discloses an ex vivo method for producing a preparation containing CD4+ T cells specific for EBV structural antigens for use in the prophylaxis and treatment of patients with a reduced T cell activity in order to prevent or treat growth of EBV infected B cells.

Abstract: A continuous flow bioreactor system that includes a bioreactor, an optional post-bioreactor preparation chamber, a cell sorter, and an optional pre-bioreactor preparation chamber in a closed loop, useful for enriching a heterogeneous cell population growing in the bioreactor with an isolated subpopulation of cells.

Abstract: The present invention provides a mechanism for obtaining an almost unlimited source of stem cells. A method for the expansion of a population of stem cells that retain their developmental capacity comprises co-culture of a stem cell population with a trophic support, for example, endothelial cells or conditioned medium derived from endothelial cell culture. The method also provides a method for enhancing neurogenesis when neural stem cells are co-cultured with the endothelial cell-derived trophic support to retain their neurogenic potential.

Abstract: An object of the invention is to provide artificial skin that does not contain any animal-derived material or pathogen and has excellent biocompatibility. The invention provides, as a solution means, a method for producing artificial skin comprising the steps of: (A) forming a dermal layer by solidifying a mixture of dermal fibroblasts and a peptide hydrogel having a fibrous structure; and (B) forming an epidermal layer by seeding skin keratinocytes onto the dermal layer obtained in Step (A), and culturing the epidermal keratinocytes.

Abstract: The present invention relates to methods and kits for expanding a stem cell population. More particularly, the invention relates, inter alia, to methods, kits, and compositions for expanding a stem cell population, particularly a hematopoietic stem cell population.

Abstract: The invention relates to a purified, novel anti-inflammatory population of macrophage and methods of making and using such macrophage.

Abstract: Disclosed is a method and compositions for the differential expansion of fetal cells over maternal cells. In the method, cells from a sample of maternal blood containing CD34+ cells of both maternal and fetal origin are incubated in the presence of Stem Cell Factor in serum free media. It has been discovered that incubation of fetal cells in the presence of SCF will preferentially expand the fetal cells relative to adult cells. Fetal cells can also be identified, enriched or obtained by differential expansion of the fetal cells during colony formation. It has been discovered that differential expansion of fetal cells can result in colonies of fetal cells that are larger than colonies of adult cells. The fetal CD34+ cells can be expanded without generation of significant clonal genetic artifacts during expansion. Also disclosed is a method and compositions for producing differentiated fetal cells.

Abstract: Disclosed are methods for detecting antibody in a sample, where the antibody targets an antigen expressed by red blood cells or red blood cell ghosts. Rather than detecting the binding events between a particular antigen antibody pair (as in traditional agglutination based assays) the methods herein allow for multiplexed detection of clinically important allo-immune antibodies to blood group antigens. Specifically the method involves generating fluorescently encoded red blood cells or red blood cell ghosts with known antigen presentation and using them to detect the presence of antibody in serum/plasma with a fluorescent sandwich type immunoassay. The assay results can be read using flow cytometric or fluorescent microscope based imaging techniques.

Abstract: A production method of T cells is disclosed which includes generating iPS cells from immune cells and differentiating the iPS cells into desired immune cells. In this method, 4 different genes Oct4, Sox2, Klf4 and c-Myc are introduced into immune cells for generation of iPS cells, and the iPS cells are then differentiated into immune cells by coculture with OP9 cells. Source immune cells are taken from a patient, and the produced desired immune cells are injected into the patient for medical treatment.

Abstract: An article is provided herein for use in seeding cells on at least one filter extending across at least one well of an assay device. The article includes an elastomeric body having spaced-apart first and second surfaces, and at least one channel extending between, and through, the first and second surfaces. The channel is formed to sealingly, and detachably, engage an outer surface of the well with the filter being at least partially encompassed by the channel. Advantageously, with the subject invention, a cell monolayer can be formed on the exterior surface of the filter. The assay device may be a multiwell plate, an insert plate, a column, a test tube, or a pipette.

Abstract: A method for providing a urinary tract tissue graft composition includes providing a segment of small intestinal submucosa and isolating and culturing at least one multipotent cell type from a tissue specimen of a subject. The at least one multipotent cell type is then seeded upon a surface of the segment of small intestinal submucosa and allowed to differentiate into a unipotent cell type, thereby forming a urinary tract tissue graft. Methods for repairing a damaged urinary tract tissue of a subject are also disclosed.

Abstract: Disclosed are various methods and bioreactor devices for culturing retinal cells and/or tissues. The bioreactor devices may, in certain embodiments, include a microchannel network, a scaffold for culturing neuroretinal cells, and a porous membrane separating the microchannel network from the scaffold.

Abstract: The present invention relates to methods of constructing an integrated artificial immune system that comprises appropriate in vitro cellular and tissue constructs or their equivalents to mimic the normal tissues that interact with vaccines in mammals. The artificial immune system can be used to test the efficacy of vaccine candidates in vitro and thus, is useful to accelerate vaccine development and testing drug and chemical interaction with the immune system.

Abstract: A process for producing a tissue transplant construct for reconstructing a human or animal organ. The process may include the steps of: (a) isolation and two-dimensional cultivation of organ-specific tissue cells; (b) application of the organ-specific tissue cells to a biocompatible, collagen-containing membrane; and, (c) cultivation of the organ-specific tissue cells on the membrane with biochemical and mechanical stimulation of the organ-specific tissue cells. Tissue transplant constructs and methods for using tissue transplant constructs are also taught.

Abstract: Process of incorporating active and aggressive molecules into a microalga, with targeted recognition of cancer cells. The invention relates to the incorporation of active molecules into a microalga having first developed a plasmid, and/or a gene with resistance to the specific molecules. The process of the invention is particularly intended for human and veterinary medicine.

Abstract: The invention relates to the direct selection of metabolic pathways having a determined function in the transformation of a substrate {Ai} into a target product {B}, which is of interest in the industrial, pharmaceutical or agri-food sectors. More specifically, the invention relates to the detection, within metagenomic libraries, of novel biosynthesis pathways involved in a biochemical reaction having a known product {B}. The selection and characterization of said novel metabolic pathways enables {B} to be produced enzymatically. The invention provides an alternative to the chemical synthesis of the molecule in question {B}. Moreover, and above all, the invention can be used specifically to target and exploit the only metabolic pathways enabling the transformation of {Ai} into {B}, while eliminating the associated metabolic pathways that can catabolise the target product {B}.

Abstract: The invention provides for a method for aggregating dermal papilla cells or dermal sheath cells or a combination thereof, the method comprising: growing dermal papilla cells or dermal sheath cells or a combination thereof in suspension culture; and contacting the culture with an effective amount of an enzyme, wherein a substrate of the enzyme is an extracellular matrix molecule in the suspension culture, so as to aggregate dermal papilla cells or dermal sheath cells. The culture may be a hanging drop culture and the enzyme may be a hyaluronidase.

Abstract: Methods of inhibiting proliferation of a plurality of proliferating cells are disclosed. Methods of inhibiting cell overgrowth on compositions that are in an animal's body are disclosed. Methods of inhibiting cell overgrowth on a device that is in an animal's body are disclosed. Devices that have on their exterior surface an alginate matrix that comprises Strontium are disclosed. Compositions comprising an alginate body and alginate sheets that each comprise a single layer of cells coating the exterior surface of the alginate body are disclosed. Methods of preparing an artificial tissue are disclosed. Devices comprising cells encapsulated within an alginate matrix and/or maintained as a monolayer on an alginate body, and methods of making and using the same are disclosed. Methods of coating compositions and devices are disclosed.

Abstract: A new method for selecting clones and recloning mammalian cells which are of importance for the production of biopharmaceuticals, preferably hamster or mouse myeloma cells, with a high degree of automation and throughput. The invention relates to methods of depositing and replicating single cell clones of the cells in question. The invention also relates to methods of preparing proteins using cells which have been obtained and replicated by single cell deposition as well as compositions which allow the replication of single cells.

Abstract: The invention relates to a method for infiltration of polypeptides in cells. The invention further relates to the use of the cells and urea-adjuvated polypeptides for the diagnosis, treatment or prevention of diseases. The invention further relates to the detection of polypeptide-specific immune cells.

Abstract: Disclosed is a method for in vitro growing of connective tissue substitute, said connective tissue substitute being populated with fibroblasts, a connective tissue substitute obtainable by such a method, as well as a method for closing of a wound, wherein such connective tissue substitute is applied onto a wound.

Abstract: The invention relates to a method of excysting and growing protozoal oocysts by in vitro tissue culture resulting in production of a continuous culture of merozoites. The invention also provides an economical and reliable supply of cultured Eimeria sp. for vaccine production, assays and research. Domesticated avians that have been vaccinated using the provied Eimeria sp. are also provided.