Abstract: The present invention relates to methods for preparing an artificial immune system. The artificial immune system comprises a cell culture comprising T cells, B cells and antigen-primed dendritic cells. The artificial immune system of the present invention can be used for in vitro testing of vaccines, adjuvants, immunotherapy candidates, cosmetics, drugs, biologics and other chemicals.

Abstract: Compositions and methods for creating a laminar construct for tissue-engineered dermal equivalent are provided. One composition provided herein comprises a hydrogel matrix comprising two or more hydrogels layers and a population of stem cells. Associated methods are also provided.

Abstract: Provided are methods of the production of patterned 3-dimensional biopolymer scaffolds containing living cells. The methods include selective photopolymerization of biopolymers to create patterned structures and the patterning of cells within relatively homogenous slabs of biopolymer using dielectrophoresis. Also provided are patterned 3-dimensional biopolymer scaffolds generated by the methods and their use.

Abstract: The present invention features microgels and microtissues for use in tissue engineering. Featured is a microencapsulation device for making microgels and/or microtissues via an emulsion technology. Also featured are methods of making higher ordered structures that mimic in vivo tissue structures. Methods of us are also featured.

Abstract: A microfiber showing improved mechanical strength, which comprises a micro gel fiber consisting of collagen gel or the like covered with high strength hydrogel such as alginate gel.

Abstract: A microfluidic device for culturing cells, termed a microscale cell culture analog (?CCA), is provided. The microfluidic device allows multiple cell or tissue types to be cultured in a physiologically relevant environment, facilitates high-throughput operation and can be used for drug discovery. The microfluidic device uses gravity-induced fluidic flow, eliminating the need for a pump and preventing formation of air bubbles. Reciprocating motion between a pair of connected reservoirs is used to effect the gravity-induced flow in microfluidic channels. Bacterial contamination is reduced and high throughput enabled by eliminating a pump. The microfluidic device integrates a pharmacokinetic-pharmacodynamic (PK-PD) model to enable PK-PD analyses on-chip. This combined in vitro/in silico system enables prediction of drug toxicity in a more realistic manner than conventional in vitro systems.

Abstract: The invention relates to the use of encapsulates of cancer cells, in agarose coated, agarose containing beads, for isolating chemotherapeutic resistant cells which have at least one stem cell property, such as expression of OCT4. The cells thus isolated are also a feature of the invention, as is a method for screening for potential therapeutic agents.

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: Provided herein are microfluidic devices that can be used as a 3D bioassay, e.g., for drug screening, personalized medicine, tissue engineering, wound healing, and other applications. The device has a series of channels {e.g., small fluid channels) in a small polymer block wherein one or more of the channels can be filled with a biologically relevant gel, such as collagen, which is held in place by posts. As shown herein, when the device is plated with cells such as endothelial cells, new blood vessels grow in the gel, which is thick enough for the cells to grow in three dimensions. Other channels, e.g., fluid channels, allow drugs or biological material to be exposed to the 3D cell growth. Cells, such as endothelial cells, can be cultured and observed as they grow on the surface of a 3D gel scaffold, where e.g., rates of angiogenesis can be measured, as well as intervascularization and extravascularization of cancerous cells.

Abstract: The present disclosure provides compositions comprising musculoskeletal cells and mesenchymal stem cells in discrete regions. The present disclosure provides systems comprising a subject composition; and methods of using a subject composition to generate cartilage, bone, tendon, muscle, intervertebral disc, or other musculoskeletal tissues.

Abstract: The present invention relates to a method and to vectors for the immortalisation of cells independent of their type. It further relates to a cell or a cell line produced with the method or the vectors of the invention. The invention also relates to the use of this cell or cell line in in vitro applications and in the treatment of disease.

Abstract: Alginate polyelectrolyte encapsulation is used for the controlled differentiation of embryonic stem cells. An isolated cell population is provided. The cell population includes a single cell suspension of ES cells encapsulated within an alginate polyelectrolyte microenvironment. The encapsulated ES cells are capable of differentiating within said microenvironment into hepatocyte lineage cells in the absence of embryoid body intermediates or growth factor supplementation.

Abstract: In a method of forming a cellular structure, cells and a transient linker are supplied to a volume partially enclosed by a cage. The linker facilitates initial attachment of adjacent cells to form a cell aggregate. The cage defines distributed openings that are sized to retain the cell aggregate. A fluid comprising a cell culture medium is supplied to the volume. The fluid is withdrawn from the volume through the openings. Aggregated cells retained in the volume are cultured to form a cell structure. A cell culturing device is provided which comprises a conduit and a cage in the conduit. A fluid flows in the conduit. The fluid comprises the cells, the transient linker and the cell culture medium. The cage retains aggregated cells formed in the fluid, and defines distributed openings that allow the fluid to flow through.

Abstract: Provided is a method for the derivation of neural stem cells (NSCs) from embryonic stem cells (ESCs) and the use of the NSCs for treatment of various neural disorders. The NSCs that are derived from the ESCs are tissue-specific multipotent NSCs with a stable growth rate, unlimited self-renewal capacity, and a predictable differentiation profile. Being both non-tumorigenic and engraftable, the NSCs of the present invention have utility in repopulation stroke-damaged tissue. The NSCs of the present invention may be differentiated to produce tyrosine-hydroxylase expressing neurons, which may be used as a source of dopaminergic neurons for subjects suffering from a condition characterized by dopaminergic dysfunction, such as Parkinson's disease.

Abstract: A microfiber showing improved mechanical strength, which comprises a micro gel fiber consisting of collagen gel or the like covered with high strength hydrogel such as alginate gel.

Abstract: There is provided a method of forming a hydrogel, the method comprising: providing a mixture of a polymer comprising a cross-linkable pendant phenolic group, peroxidase, H2O2, fibrinogen, and thrombin, at concentration sufficient to enzymatically cross-link the polymer and to cleave the fibrinogen to yield fibrin; and allowing the mixture to form a hydrogel. There is also provided a hydrogel comprising a cross-linked network of a polymer interpenetrated by fibrin fibers, the polymer cross-linked by oxidative coupling between phenolic groups pendant on the polymer.

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 tissues of the immune system in mammals. The artificial immune system can be used to test the efficacy of vaccine candidates and other materials in vitro and thus, is useful to accelerate vaccine development and testing drug and chemical interactions with the immune system, coupled with disease models to provide a more complete representation of an immune response.

Abstract: The invention relates to a microtiter plate and use thereof for conducting fermentation under fed-batch conditions. In order to produce a microtiter plate which permits screening under fed-batch conditions, the invention proposes that the cavities (2) of the microtiter plate according to the invention be filled with a culturing fluid and nutrient solution and be designed in such a way that each of the cavities (2) of the microtiter plate which is filled with nutrient solution is connected by a channel (4) to at least one other further cavity (3) of the microtiter plate which is filled with a culturing fluid. A diffusion barrier (13) arranged in the material permeable channel (4) controls the kinetics of the material transfer of nutrients from the cavity containing the nutrient solution to the cavities containing the culturing fluid.

Abstract: The present invention relates to a microsphere-containing cell aggregate including: hydrogel microspheres being obtained by chemical cross-linking of one or more water-soluble synthetic macromolecules selected from the group consisting of water-soluble synthetic polymers, polysaccharides, and proteins; and cells. The present invention also relates to a method for producing the microsphere-containing cell aggregate.

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: Compositions and methods for producing encapsulated cells having an average diameter of less than about 200 ?m are provided. Methods for using the disclosed encapsulated cells are also provided.

Abstract: A method is provided to functionally select cells with enhanced characteristics relevant to cell engraftment, including both spontaneous migration and directional migration towards specific chemo-attractants. The cells are preferably undifferentiated cells, such as mesenchymal stem cells. The method involves entrapping or encapsulating the cells in a biomaterial barrier, optionally inducing cell migration, and selecting cells that migrated through the barrier. The cells selected by this method have better migratory activities and enhanced in vivo engraftment to injured tissues when they are supplemented systemically.

Abstract: A microfluidic device for culturing cells, termed a microscale cell culture analog (?CCA), is provided. The microfluidic device allows multiple cell or tissue types to be cultured in a physiologically relevant environment, facilitates high-throughput operation and can be used for drug discovery. The microfluidic device uses gravity-induced fluidic flow, eliminating the need for a pump and preventing formation of air bubbles. Reciprocating motion between a pair of connected reservoirs is used to effect the gravity-induced flow in microfluidic channels. Bacterial contamination is reduced and high throughput enabled by eliminating a pump. The microfluidic device integrates a pharmacokinetic-pharmacodynamic (PK-PD) model to enable PK-PD analyses on-chip. This combined in vitro/in silico system enables prediction of drug toxicity in a more realistic manner than conventional in vitro systems.

Abstract: The present invention relates to methods for preparing an artificial immune system. The artificial immune system comprises a cell culture comprising a three-dimensional matrix comprising lymphoid tissue, a three-dimensional matrix comprising epithelial and/or endothelial cells, and diseased cells. The artificial immune system of the present invention can be used for in vitro testing of vaccines, adjuvants, immunotherapy candidates, cosmetics, drugs, biologics and other chemicals.

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 tissues of the immune system in mammals. The artificial immune system can be used to test the efficacy of vaccine candidates and other materials in vitro and thus, is useful to accelerate vaccine development and testing drug and chemical interactions with the immune system, coupled with disease models to provide a more complete representation of an immune response.

Abstract: Described herein are composites useful in tissue and organ engineering. In one aspect, the composite comprises the reaction product between a macromolecule comprising at least one thiol group and a gold nanoparticle. The thiolated macro-molecule crosslinks with the gold nanoparticle to produce a composite that is useful in anchoring cells. The composites can be used to form multi-layer 3-D structures, where the cells in each layer can aggregate and fuse with one another to form tissues and organs.

Abstract: The present invention relates to artificial tissue growth guides comprising a core and an outer sleeve, which facilitates the regeneration of damaged tissues, such as nerves. The core is fixed to the sleeve at two attachment sites so that cells seeded within the core produce mechanical tension between the attachment sites. This tension aligns the cells and the fibres of the core and provides an improved substrate for tissue regeneration. Growth guides may be surgically implanted into an individual.

Abstract: A method of inhibiting graft rejection includes isolating Hepatic Stellate Cells from a mammal liver, activating the isolated Hepatic Stellate Cells, and administering a combination of Hepatic Stellate cells and a graft to a mammal.

Abstract: The present invention is directed to a non-immunogenic cellular composition comprising: a cell having a cell surface and antigenic determinants on the cell surface; an optional linker molecule covalently attached to the cell surface; and a hydrophilic, biocompatible, non-immunogenicity providing compound or polymer (e.g., polyethylene glycol or a derivative thereof) covalently attached to the linker molecule or directly to the cell. In one embodiment, the linker molecule is covalently attached directly to the antigenic determinant on the cell surface. In an alternate embodiment, the linker molecule may be covalently attached to a non-antigenic site on the cell surface, but will camouflage the antigenic determinant on the cell surface.

Abstract: Improved methods for tissue engineering including tissue transplantation, augmentation and regeneration are described. These methods utilize a support matrix in the generation of an anatomical construct that includes donor vascularized tissue. The support matrix may be devised such that it has size and shape adapted to simulate the dimensions of tissue to be transplanted, augmented and/or repaired.

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: The invention provides media formulations. A complete media formulation of the invention includes, for example, the following components: albumin, an iron carrier, glutamine, a glycosidase or hydrolase, fibroblast growth factor (FGF), a salt or mineral, and essential amino acids, at an osmolarity of about 220-330 mOsm/Liter.

Abstract: Compositions, methods of production and use, and kits for an osteochondral graft involving both articular cartilage and underlying bone are provided.

Abstract: Dispersions that comprise insoluble alginate/gelling ion particles in an alginate solution, wherein the dispersion exhibits less than 10% of final gel storage modulus after one minute in the absence of addition of non-gelling cations are disclosed. Kits and compositions for making such dispersions are disclosed and methods for making and using the dispersions, and the components used in the dispersions are also disclosed.

Abstract: The present invention generally relates to methods for culturing and analyzing cells using liquid bridges.

Abstract: The invention relates to the stem cells, embryonic stem cells in particular. It has been found that, when these stem cells are entrapped such that their proliferation is inhibited, they produce material which inhibits the proliferation of other, non-entrapped cells, including stem cells and neoplastic and/or hyperproliferative, but otherwise normal cells. It has also been found that entrapped cancer cells will produce material which inhibits the proliferation of stem cells. Further, it has been found that the entrapment of the stem cells inhibits their differentiation and thus the entrapment process can serve as a long-term storage device for maintaining the undifferentiated state of at least a portion of the entrapped 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: An apparatus for encapsulating cells is disclosed. According to one embodiment, the apparatus includes an indirect-pumping dispenser for dispensing a cell suspension into an encapsulation solution through an outlet of the dispenser. The apparatus also includes a dipping mechanism that is attachable to the outlet of the dispenser. The dipping mechanism is adapted to dip the dispenser outlet in the encapsulation solution to allow the cell suspension dispensed thereat to come into contact with the encapsulation solution.

Abstract: The invention provides methods for expanding mesenchymal stem cells (MSCs) in non-adherent cultures. The methods include the propagation of MSCs in or on non-adherent matrices. The invention further provides administration and the use of cells propagated by the method of the invention for administration and preparation of a therapeutic agent. The invention further provides kits including cells propagated by the methods of the inventions.

Abstract: Alginate polyelectrolyte encapsulation is used for the controlled differentiation of embryonic stem cells. An isolated cell population is provided. The cell population includes a single cell suspension of ES cells encapsulated within an alginate polyelectrolyte microenvironment. The encapsulated ES cells are capable of differentiating within said microenvironment into hepatocyte lineage cells in the absence of embryoid body intermediates or growth factor supplementation.

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: Provided is a method and system for the rapid and accurate detection of growth and metabolism of a cellular microorganism in a population of microorganisms in a non-liquid, culture medium. Further provided is a gelled culture medium containing a non-toxic, water-soluble, phosphorescent compound which measures oxygen content (partial pressure) of an microorganism also contained therein, by oxygen-dependent quenching of phosphorescence; or the gel contains a fluorescent pH indicator that demonstrates growth of the microorganism by pH-dependent intensity change or wavelength shift in the emission spectrum. Further provided is a system and method for killing undesirable microorganisms or colonies in the culture medium without harming the surrounding microorganisms.

Abstract: The present inventors identified aggregation of embryonic stem cells and embryoid bodies (EBs) as the cause of the difficulty in generating large numbers of the embryonic stem cells (ES) cell-derived tissues. To counter this, the invention provides a novel bioprocess where aggregation of spheroid forming cells, such as embryonic stem cells and spheroids, such as EBs is controlled, such as by encapsulation of within a matrix. As a result, EBs can be generated with high efficiency and cultured in high cell density, well-mixed systems. Well-mixed conditions facilitate measurement and control of the bulk media conditions and allow for the use of scalable bioreactor systems for clinical production of tissue. Therefore, the invention enables generation of ES cell-derived tissue on a clinical scale. The invention is also applicable to any spheroid-forming cells and other types of pluripotent cells.

Abstract: Method of providing a substrate with a ready-to-use uniformly distributed extracellular matrix is disclosed. This method includes applying extracellular matrix components to a substrate area; incubating the extracellular matrix components to allow polymerization thereof; freezing the polymerized extracellular matrix on the substrate area; lyophilizing the frozen extracellular matrix on the substrate area; and allowing the lyophilized extracellular matrix to warm to room temperature. Also contemplated is a cell culture apparatus having a dried uniformly distributed extracellular matrix formed by the above-mentioned method.

Abstract: Embodiments provide cell culture methods to produce a 3-D model assembly that mimics in vitro the microenvironment of human bone marrow, where cells occupy distinct niches. Methods and apparatuses are provided for the efficient testing of cancer, including malignant hematopoietic cancers and metastatic spread to the bone marrow of solid tumors, and of other diseases of the blood and bone marrow. Methods and apparatuses are further provided in embodiments for the investigation of bone marrow cell biological characteristics and for the testing of therapeutics for nonmalignant diseases of the blood and bone marrow.

Abstract: The invention provides systems, including apparatus, methods, and kits, for the microfluidic manipulation and/or detection of particles, such as cells and/or beads. The invention provides systems, including apparatus, methods, and kits, for the microfluidic manipulation and/or analysis of particles, such as cells, viruses, organelles, beads, and/or vesicles. The invention also provides microfluidic mechanisms for carrying out these manipulations and analyses. These mechanisms may enable controlled input, movement/positioning, retention/localization, treatment, measurement, release, and/or output of particles. Furthermore, these mechanisms may be combined in any suitable order and/or employed for any suitable number of times within a system.

Abstract: The invention relates to developments in the treatment of diabetes in mammals. Particularly it relates to a method of preparing a xenotransplantable porcine islet preparation capable upon xenotransplantation of producing porcine insulin in an appropriate recipient mammal, the method including or comprising the steps of: (I) harvesting the pancreas of piglets at or near full term gestation, and (ii) extracting pancreatic ? islet cells from the harvested pancreas wherein the islets (at least at some stage in the performance of the method) are exposed to nicotinamide. Further, the invention relates to a method of encapsulation of a xenotransplantable porcine islet preparation, and transplantation of such a preparation, or a capsule containing such a preparation, into an appropriate recipient mammal.

Abstract: There is provided a method for growing human intervertebral cells. Disc tissue is surgically removed from a normal disc of a patient, the cells expanded by feeding with a cell stimulant such as a growth factor, or a cytokine or a bioactive agent to form monolayer primary cell cultures on a plastic mesh such as a nylon mesh. In the case of a growth factor, fetal bovine serum is preferred as it improves cell proliferation and production of appropriate extracellular matrix components. In another aspect of this invention, the monolayer primary cell cultures are seeded in alginate or agarose and fed again with the cell stimulant until three-dimensional cell cultures are formed. The cells are recovered from the alginate or agarose or from monolayer cultures. Re-implantation is carried out using bioresorbable carriers or cell suspensions.

Abstract: The invention relates to developments in the treatment of diabetes in mammals. Particularly it relates to a method of preparing a xenotransplantable porcine islet preparation capable upon xenotransplantation of producing porcine insulin in an appropriate recipient mammal, the method including or comprising the steps of: (I) harvesting the pancreas of piglets at or near full term gestation, and (ii) extracting pancreatic ? islet cells from the harvested pancreas wherein the islets (at least at some stage in the performance of the method) are exposed to nicotinamide. Further, the invention relates to a method of encapsulation of a xenotransplantable porcine islet preparation, and transplantation of such a preparation, or a capsule containing such a preparation, into an appropriate recipient mammal.

Abstract: Biological agents such as secretory cells are encapsulated in a hydrophilic gel made of agarose or collagen-agarose and gelatin sponge-agarose combinations. In a preferred embodiment, semi-solid beads are formed from a suspension containing collagen, agarose and secretory cells such as pancreatic islets, the collagen is polymerized to form solid, agarose-collagen beads and the solid beads are coated with agarose. Coating is preferably by rolling the solid beads in about 5-10% agarose, contacting the rolled beads with mineral oil and washing oil from the beads. Beads containing secretory cells can be transplanted into a mammal to treat a condition caused by impaired secretory cell function.