Abstract: The present invention pertains to a system for culturing cells comprising a culturing bag and a continuous flow centrifuge wherein the cells are continuously separated from the supernatant and are recycled into the culturing bag. Further provided are methods for culturing cells and for producing a biological substance using the device for culturing cells, and the use of a bag for culturing cells in said device or said methods. In particular, a perfusion system for culturing cells is provided wherein the wave technology for culturing cells is combined with continuous flow centrifugation for separating the medium from the cells.

Abstract: A disposable apparatus for cell expansion, having at least one bioreactor. The bioreactor has a cellular growth area and a supply area, the cellular growth area being separated from said supply area by a membrane. A fluid recirculation path in fluid communication with the cellular growth area allows for hermetically removing a sample containing cellular matter. This may comprise an elongated tube, or a plurality of parallel tube segments. The parallel tube segments have inflow ends and outflow ends, and the inflow ends are joined at a first common juncture and the outflow ends are joined at a second common juncture. The common junctures may comprise valves.

Abstract: Gas permeable devices and methods are disclosed for cell culture, including cell culture devices and methods that contain medium at heights, and certain gas permeable surface area to medium volume ratios. These devices and methods allow improvements in cell culture efficiency and scale up efficiency.

Abstract: Gas permeable devices and methods are disclosed for cell culture, including cell culture devices and methods that contain medium at heights, and certain gas permeable surface area to medium volume ratios. These devices and methods allow improvements in cell culture efficiency and scale up efficiency.

Abstract: Provided are means and methods for producing embryoid bodies (EBs) from multi- or pluripotent cells. In particular, a method of generating embryoid bodies (EBs) is described comprising agitation of a liquid suspension culture of multi- or pluripotent cells in a container until generation of cell aggregates, optionally diluting the suspension, and further agitation of the suspension until formation of EBs. Furthermore, the present invention relates to the use of the novel culturing method and EBs obtained thereby for a variety of applications including genomics, diagnostic assays, teratogenic/embryotoxicological and pharmacological assays as well as for the provision of tissue grafts.

Abstract: A carrier for cell culture is provided which improves the cell proliferativity in serum-free culture and which is free from risk from infection factor contamination. The gist of the features of the present invention is to be formed of a crosslinked poly(meth)acrylic acid (salt) particle (A) and an artificial polypeptide (P) having at least one cell-adhesive minimal amino acid sequence (X) in one molecule and to have a water retention value of from 2 to 50 g/g. The (A) is preferably a particle produced by reversed phase suspension polymerization of an aqueous monomer solution containing (meth)acrylic acid and/or an alkali metal salt of (meth)acrylic acid. The (P) preferably has at least one auxiliary amino acid sequence (Y) in one molecule of the (P). The (X) is preferably an Arg Gly Asp sequence.

Abstract: Motor neuron progenitor (MNP) cells and populations of MNP cells, are provided, in particular, populations of human late stage MNP cells having a purity of greater than about 65% late stage MNP cells and high-purity populations of MNP cells having greater than 95% viable cells, as well as method of making and using the same, including deriving late stage MNP cells from pluripotent embryonic stem cells, producing high-purity populations of late stage MNP cells, producing populations of viable MNP cells, transporting viable MNP cells, and transplanting MNP cells.

Abstract: A method is provided to prepare a plurality of microwells suitable for the formation of embryoid bodies from embryonic stem cells. The method requires applying an image-forming material to a heat sensitive thermoplastic material in a designed pattern and heating the material under conditions that reduce the size of the receptive material by at least about 60% to create a mold. A polymer such as PDMS is then applied to the mold and removed to form the microwells. In an alternative aspect, the plurality of microwells on receptive material are prepared by etching a microwell designed pattern into a heat sensitive thermoplastic material support and then heating the material under conditions that reduce the size of the receptive material by at least about 60%.

Abstract: The invention relates to a process for the culturing of cells, preferably El-immortalized HER cells, more preferably PER.C6 cells in a reactor in suspension in a cell culture medium, wherein the cells produce a biological substance, preferably an antibody, wherein at least one cell culture medium component is fed to the cell culture and wherein the cell culture comprising the cells, the biological substance and cell culture medium is circulated over a separation system and wherein the separation system separates the biological substance from substances having a lower molecular weight than the biological substance and wherein the biological substance is retained in or fed back into the reactor. Preferably part of the substances of lower molecular weight is continuously removed from the cell culture.

Abstract: An improved monocyte activation test is described that is better able to detect non-endotoxin pyrogens in medical products, in which a sample is incubated with a monocyte-containing reagent in an assay system comprising at least one surface comprising polypropylene. The invention also concerns assay systems for use in these tests that include at least one microtiter well having at least one interior surface comprising polypropylene and having a shape such that monocyte-containing reagent is concentrated in the well to provide greater cell to cell contact. The invention also relates to a diagnostic kit that can be used to test for the presence of non-endotoxin pyrogens in a sample.

Abstract: A gas-introduction system useful, for example, in biotechnology for supplying cells or microorganisms with oxygen. The gas-introduction system has a bubble column and a distributor. In the vessel there is situated a liquid medium which is to be supplied with gas. A vector is used as transport medium for the gas. The gas is introduced into the bubble column and here taken up by the vector. The vector is applied to the liquid surface via the distributor in droplet form, falls downward in the medium and releases to the medium some of the gas that has been taken up. At the bottom of the vessel is situated a collecting device in which the vector droplets coalesce and pass back into the bubble column. Disclosed are a bioreactor comprising the gas-introduction system and a method for introducing gas into a liquid medium, preferably an aqueous suspension containing cells or microorganisms.

Abstract: Co-cultures of heterogeneous cell populations in a diffusion-constrained microenvironment and methods for co-culturing are disclosed.

Abstract: The invention relates to a microfluidic device comprising one or more fluid channels, one or more fluid ports, and a V-shaped particle retention structure. The fluid channel is generally opposite the particle retention structure, fluid ports are located between the fluid channel and the particle retention structure, and the particle retention structure has sloped side walls. Fluid, including reagents, can be delivered to the microfluidic device through the one or more fluid channels or the fluid ports. The invention also relates to methods of using the microfluidic device to monitor, observe, measure, or record a biological parameter of a particle, to separate a single particle from a group of particles, to culture a cell, to treat a particle, and to move a particle back and forth in the device.

Abstract: The invention relates to an impeller for growing adherent mammalian cells on microcarrier beads. In other aspects, the invention relates to methods of using the impeller and to bioreactors or cell separators comprising the impeller.

Abstract: A bioreactor includes a cylinder, a mandrel and a filter film. The cylinder includes two end walls and a circumferential wall interconnected with the both end walls; the two end walls and circumferential wall define together a reaction chamber. The mandrel is disposed between the two end walls. The mandrel has an inlet path and an outlet path provided on its both ends; the filter film wraps the mandrel to prevent passage of the first kind of substance while permit the passage of the second kind of substance; a gap is defined between the filter film and mandrel; at least one portion of the filter film is tied so as to divide the gap into multiple separate and isolated gap regions to prevent the second fluid from direct exiting from the outlet path through the gap. Also disclosed are control system and method for the bioreactor.

Abstract: The invention relates to a process for the culturing of cells, preferably E1-immortalized HER cells, more preferably PER.C6 cells in a reactor in suspension in a cell culture medium, wherein the cells produce a biological substance, preferably an antibody, wherein at least one cell culture medium component is fed to the cell culture and wherein the cell culture comprising the cells, the biological substance and cell culture medium is circulated over a separation system and wherein the separation system separates the biological substance from substances having a lower molecular weight than the biological substance and wherein the biological substance is retained in or fed back into the reactor. Preferably part of the substances of lower molecular weight is continuously removed from the cell culture.

Abstract: Methods of forming soft connective tissue compositions such as skin equivalents, compositions made by the methods and their uses.

Abstract: A method for determining the effect of a plurality of culture conditions on a cell, comprising the steps of: a) providing a first set of groups of cell units each comprising one or more cells, and exposing said groups to desired culture conditions; (b) pooling two or more of said groups to form at least one second pool; (c) subdividing the second pool to create a further set of groups of cell units; (d) exposing said further groups to desired culture conditions; (e) optionally, repeating steps (b)-(d) iteratively as required; and (f) optionally assessing the effect on a given cell unit of the culture conditions to which it has been exposed.

Abstract: A method for the differentiation of mammalian pluripotent stem (PS) cells into a mortal multi-lineage progenitor cell population is provided which comprises culturing the pluripotent stem cells in the presence of Hyaluronan (HA). The mortal multi-lineage progenitor cell population may be a population of mesenchymal stem cells. The mortal multi-lineage progenitor cell population may form cells of the mesodermal lineage, suitably osteoblasts. Alternatively, the mortal multi-lineage progenitor cell population may form cells of the endodermal lineage or of the ectodermal lineage, which may be neuronal progenitors.

Abstract: The present invention provides muscle-derived progenitor cells that show long-term survival following transplantation into body tissues and which can augment soft tissue following introduction (e.g. via injection, transplantation, or implantation) into a site of soft tissue. Also provided are methods of isolating muscle-derived progenitor cells, and methods of genetically modifying the cells for gene transfer therapy. The invention further provides methods of using compositions comprising muscle-derived progenitor cells for the augmentation and bulking of mammalian, including human, soft tissues in the treatment of various functional conditions, including malformation, injury, weakness, disease, or dysfunction. In particular, the present invention provides treatments and amelioration for urinary incontinence and other urinary tract pathologies.

Abstract: Compositions for promoting growth and/or differentiation of a stem cell are disclosed. The composition comprises: a) a diamond film; b) a stem cell cultured on the diamond film; and c) a medium bathing the stem cell. The stem cell may be a mammalian neural stem cell and the diamond film may comprise a hydrogen-terminated or an oxygen-terminated surface. The hydrogen-terminated surface promotes proliferation and differentiation of a neural stem cell into neurons, and the oxygen-terminated surface promotes a neural stem cell to proliferate and differentiate into oligodendrocytes.

Abstract: The present invention provides methods and compositions for the propagation and expansion of neural precursor cells (NPCs). NPCs may be used in the clinical implementation of stem cell therapy to treat disorders such as Parkinson's disease, Huntington's disease, neuropathic pain and other diseases of the central nervous system. The large-scale production of NPCs in bioreactors allows for the generation of clinical quantities of these cells.

Abstract: Disclosed are a composition for introducing the osteogenic differentiation of human embryonic stem cells and a method for differentiating human embryonic stem cells into an osteoblastic lineage by inhibiting the mTOR signaling pathway. When cultured in the presence of an inhibitor of the mTOR signaling pathway, human embryonic stem cells are effectively induced to differentiate into an osteoblastic lineage. The osteogenic differentiation of human embryonic stem cells using the method and the composition is useful in examining the development and differentiation mechanism of osteoblasts and the cause of metabolic bone diseases, including osteoporosis. In addition, the method and the composition can be applied to the development of osteogenic differentiation techniques for generating clinically useful, terminally differentiated mature cells or progenitor cells.

Abstract: A method for obtaining a response of a tissue model system to an activator includes contacting a bio-artificial tissue model system with an activator and measuring cellular mechanical response thereto of at least one of contractile force and tissue stiffness. A method for obtaining a response of a tissue model system to an activator includes contacting a bio-artificial tissue model system with an activator and measuring cellular mechanical response thereto of at least one of contractile force and hysteresis.

Abstract: The present disclosure describes mammalian pluripotent embryonic-like stem cells (ELSCs) isolated from corneal limbal tissue, a non-embryonic tissue. The ELSCs of the present disclosure are capable of proliferating in an in vitro culture, maintain the potential to differentiate into cells of endoderm, mesoderm, and ectoderm lineage in culture, and are capable of forming embryoid-like bodies when placed in suspension culture. Thus, these cells possess multi-lineage differentiation potential and self-renewing capability. ELSCs may be a promising therapeutic tool, and may provide new therapeutic alternatives for various diseases, conditions, and injuries.

Abstract: Disclosed herein are bioreactors including a first sheet and a second sheet (one or both of which is substantially transparent to light), wherein the second sheet is disposed adjacent to the first sheet, and the first and second sheets are sealed along a first longitudinal edge, a second longitudinal edge, a first horizontal edge, a second horizontal edge, and at least one intermediate horizontal seal between the first horizontal edge and the second horizontal edge, thereby forming at least two chambers for holding fluid in series along a vertical axis, wherein each of the two or more chambers is oriented at an angle relative to the vertical axis, wherein the angle is about 0° to about 90° and at least one of the chambers is oriented at an angle greater than 0°, and wherein there is at least one opening in each of the first horizontal edge, the second horizontal edge, and intermediate horizontal seal(s); a support structure comprising at least one horizontal support, wherein the horizontal support is located

Abstract: The present invention relates to a method for determining the ideal time for and outcome of reproductive health procedures including in vitro fertilization by establishing a correlation between the successful outcome of said procedure and the spectra of a body fluid obtained using a chosen analytical modality for a population of patients, acquiring for a patient a spectrum of the body fluid of the patient using said chosen modality.

Abstract: A polysaccharide comprising galactose, glucose, rhamnose, and pyruvic acid as constituents, wherein the galactose, glucose, and rhamnose are contained in a molar ratio of 4:2:1, and the pyruvic acid is contained in an amount of 4 to 7 wt %. The polysaccharide can be obtained by culturing Bifidobacterium longum JBL05 (NITE BP-82) under anaerobic conditions.

Abstract: Compositions and methods for preparing Factor IX, Factor IX-containing fusion proteins, and Factor IX-containing conjugates with processing of Factor IX propeptide by PC5, are provided. In one embodiment PC5 is used to process a precursor polypeptide for a Factor IX-Fc monomer-dimer hybrid.

Abstract: The present invention is directed to pluripotent fetal stem cells derived from chorionic villus, amniotic fluid, and placenta and the methods for isolating, expanding and differentiating these cells, and their therapeutic uses such as manipulating the fetal stem cells by gene transfection and other means for therapeutic applications.

Abstract: The present invention relates to methods for production of undifferentiated or differentiated embryonic stem cell aggregate suspension cultures from undifferentiated or differentiated embryonic stem cell single cell suspensions and methods of differentiation thereof.

Abstract: Disclosed is a culture medium for culturing an animal cell, which is characterized by containing a peptide comprising, as a constituent unit, an amino acid residue selected from the amino acid group consisting of serine, tyrosine and cysteine. The culture medium is suitable for the high level production of a protein by an animal cell.

Abstract: This document provides methods and materials related to induced pluripotent stem cells. For example, induced pluripotent stem cells, compositions containing induced pluripotent stem cells, methods for obtaining induced pluripotent stem cells, and methods for using induced pluripotent stem cells are provided. In addition, methods and materials for using induced pluripotent stem cells to repair tissue (e.g., cardiovascular tissue) in vivo as well as methods and materials for using induced pluripotent stem cells to assess their therapeutic potential in appropriate animal models are provided.

Abstract: The present invention pertains to an apparatus for holding cells. The apparatus comprises a mechanism for incubating cells having a dynamically controlled environment in which the cells are grown, which are maintained in a desired condition and in which cells can be examined while the environment is dynamically controlled and maintained in the desired condition. The apparatus also comprises a mechanism for determining the state of the cells. The determining mechanism is in communication with the incubating mechanism. The present invention pertains to a method for holding cells. The method comprises the steps of incubating the cells in a dynamically controlled environment which is maintained in a desired condition and in which the cells can be examined while the environment is dynamically controlled and maintained in the desired condition. Additionally, there is the step of determining the state of the cells.

Abstract: The disclosure provides a method of reducing or eliminating Neu5Gc in a cell culture or in a human subject. The method includes flooding the system with the human sialic acid i\7-acetylneuraminic acid (Neu5Ac) in glycosidically-bound or free form, or its precursor N-acetylmannosamine (ManNAc) in an amount sufficient to metabolically compete out the Neu5Gc, either as it enters the cells for the first time or when it recycles from break-down of preexisting cellular molecules. Additionally, Neu5Ac feeding results in reduction of Neu5Gc expression even in some animal cells capable of Neu5Gc production.

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: A device is disclosed for culturing cells in at least one cell culture well. The device includes one or more interconnecting layers having a pattern therein, the pattern including at least one microfluidic channel, at least one cell culture well having an opening at one end and a side wall, the at least one microfluidic channel in fluid communication with the side wall of the at least one cell culture well, and having a maximum channel width substantially less than a maximum width of the at least one cell culture well. The device includes at least one of a controllable valve and a controllable pump in fluid communication with the microfluidic channel, the valve and pump being configured to selectably restrict fluid transport through the microfluidic channel. In some embodiments, the device includes a removable top layer adapted to cover each of the at least one cell culture well.

Abstract: The present invention relates to a method for culturing tenocytes. In particular the present invention relates to a method for culturing tenocytes comprising the step of incubating tenocytes in a culture medium comprising insulin or functional derivative.

Abstract: This invention relates to a novel method of culturing coral tissues and polyps in vitro. Coral tissues obtained by the method of the invention may be maintained as heterotypic spheroid tissue balls for a period of at least three months or they may be induced to undergo development into new polyps, a process termed re-morphogenesis. This method can produce genetic clones of model species from single individuals that can be propagated either as undifferentiated tissue calli or as developed polyps. The products of the invention are of value to a number of educational, scientific, and commercial endeavors. Specifically, this method can be used to propagate genetic clones (strains) of a model organism for scientific research, to serve as ‘pro-environmental conservation’ sources of coral stock for educational specimens as well as a rapidly generated inventory for commercial aquarium industry.

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 concerns a method for determining an allergic response by determining the extent of degranulation of human IgE sensitized cells upon activation by allergens in food products.

Abstract: The isolation and identification of glycosaminoglycans capable of binding to proteins having a heparin-binding domain is disclosed, as well as the use of the glycosaminoglycans isolated in the growth and/or development of tissue.

Abstract: The present invention provides for methods and devices suitable for producing a transplantable cellular suspension of living tissue suitable for grafting to a patient. In applying the method and/or in using the device, donor tissue is harvested, subjected to a cell dissociation treatment, cells suitable for grafting back to a patient are collected and dispersed in a solution that is suitable for immediate dispersion over the recipient graft site.

Abstract: Solutions and suspensions comprising polymerized hemoglobin derived from human blood are disclosed. The solutions and suspensions may comprise cell culture medium, an enzyme (such as a protease), and/or a buffer. Processes of preparing the solutions and suspensions are also disclosed. The solutions and suspensions may be employed in methods of isolating mammalian cells, such as pancreatic islets, methods of preserving mammalian tissue and organs, methods of aiding the recovery of mammalian cells following their isolation, methods of maintaining mammalian cells, methods of propagating mammalian cells, and methods of treating a mammal with diabetes.

Abstract: We disclose a particle comprising a matrix coated thereon and having a positive charge, the particle being of a size to allow aggregation of primate or human stem cells attached thereto. The particle may comprise a substantially elongate, cylindrical or rod shaped particle having a longest dimension of between 50 ?m and 400 ?m, such as about 200 ?m. It may have a cross sectional dimension of between 20 ?m and 30 ?m. The particle may comprise a substantially compact or spherical shaped particle having a size of between about 20 ?m and about 120 ?m, for example about 65 ?m. We also disclose a method of propagating primate or human stem cells, the method comprising: providing first and second primate or human stem cells attached to first and second respective particles, allowing the first primate or human stem cell to contact the second primate or human stem cell to form an aggregate of cells and culturing the aggregate to propagate the primate or human stem cells for at least one passage.

Abstract: The present invention relates to a method for generating a novel form of life comprising the steps consisting of: a) irreversible alteration of the genome of a microbial clone; b) cultivation of a vast population of microbial cells originating from the altered clone obtained in step a) during numerous generations under conditions allowing selection for a higher and stable proliferation rate; c) isolation of descendant clones within the cultivated population of step b) still bearing the alteration of step a).

Abstract: Methods of culturing cells capable of producing desired proteins to obtain the proteins by use of a medium from which biological components are excluded as much as possible are provided. Specifically, a culture method characterized by culturing while maintaining a specific amino acid in a culture solution at a high concentration, and a cell culture fed-batch medium for use in the method are provided.

Abstract: In the present invention cells are placed in a multiwell plate and grown. When the assay is to be performed, one uses gravity to wash away any unbound ligands rather than vacuum or centrifugation. The cells are then examined to detect the bound ligand. To perform the washing step(s) the plate is placed into a carrier plate having open wells in register with the wells of the filter plate or one may use a wicking device or an underdrain attached to the bottom of the filter plate. Sufficient wash liquid is added to allow for filtration by the effect of gravity to occur. Cells are retained within the wells at a rate of 4 times that of other rapid methods.

Abstract: The present invention is of a method of dynamically generating human embryoid bodies which can be used for generating lineage specific cells and cell lines. Specifically, the present invention can be used to generate ESC-differentiated cells for cell-replacement therapy.

Abstract: Improvements in plasmid DNA production technology are needed to insure the economic feasibility of future DNA vaccines and DNA therapeutics. General methods are described, by means of which it is possible to dramatically increase plasmid DNA productivity in a fermentor. These processes feature Fed-batch fermentation strategies, combined with novel growth and induction phase temperature shifts.