Abstract: A multiport container system for use in cell culture processes in the field of regenerative medicine is disclosed. Specifically, the container system can have a port that connects to a fill tubing for media and reagent addition, and additional ports for tubing and connectors which are compatible to various types of cell therapy bioreactors, single use bags, as well as different sampling and cell culture systems. The multiport container system can be used for cell production processes where a single bag could be used to fill, transport, and store media or reagents, as well as to seed and feed cells into different types of culture platform. The disclosed multiport container system can be used for the streamlining of process work flow which enable rapid process development and process transfer into cell product manufacturing.

Abstract: Systems and methods for producing and using a body having a first structure defining a first chamber, a second structure defining a second chamber, a membrane located at an interface region between the first chamber and the second chamber to separate the first chamber from the second chamber. The first chamber comprises a first permeable matrix disposed therein and the first permeable matrix comprises at least one or a plurality of lumens each extending therethrough, which is optionally lined with at least one layer of cells. The second chamber can comprise cells cultured therein. The systems and methods described herein can be used for various applications, including, e.g., growth and/or differentiation of primary cells, and/or simulation of a microenvironment in living tissues and/or organs (to model physiology or disease states, and/or to identify therapeutic agents). The systems and methods can also permit co-cultures of two or more different cell types.

Abstract: Apparatuses and methods are described for the use of optically driven bubble, convective and displacing fluidic flow to provide motive force in microfluidic devices. Alternative motive modalities are useful to selectively dislodge and displace micro-objects, including biological cells, from a variety of locations within the enclosure of a microfluidic device.

Abstract: The invention relates to a system of fluidic valves and pumps and associated fluidic channels integratable into a bio-object microfluidics module. The module includes input and output buses; upstream and downstream interconnection bus control valves (CVs) coupled to the input and output buses, respectively. It may include arterial, venous, wash and waste bus lines, each connecting between the upstream and downstream interconnection bus CVs. It may also include an input CV connecting to the arterial bus line, upstream interconnection bus CV, bio-object and inlets, and an output CV connecting to the bio-object, input CV, downstream interconnection bus CV and outlets; and a pump connecting between the input CV and bio-object. The system of fluidic valves and pumps can be arranged to provide MicroFormulator functionality enabling precise mixtures of drugs, chemicals, or biochemicals to be delivered in a time-dependent fashion to biological entities housed in individual wells or chambers.

Abstract: Provided is a culture system comprising a base comprising a multiwell plate; a transwell-like type insert plate comprising a plurality of culture insert assemblies, wherein each culture insert assembly comprises a culture surface element that is removably mounted to a culture insert body; and a cover designed to fit over the transwell-like type insert plate.

Abstract: A cell culture apparatus in which on an abutting surface (2a) around an opening part of a thermostatic chamber (3) and a cold storage chamber (4) that abuts against packing elements (3b, 4b) when front doors (3a, 4a) are closed, a communication groove (5) is provided that intercommunicates the inside and outside of the thermostatic chamber (3) by a communication groove (51) when a transfer tube (U40) is removed. The shut-off mechanism (5) is formed of: a guide hole (52) that opens on the abutting surface (2a) across the communication groove (51) and is deeper than the communication groove (51); a shutter member (53) which is inserted into the guide hole (52) and moves between the blocking position blocking the communication groove (51) and the opening position opening the communication groove (51); and a spring member (54) energizing the shutter member (53) to the blocking position.

Abstract: The present disclosure provides a culture device for enumerating colonies of microorganisms. The device can comprise a base, a coversheet, and a nonporous spacer member disposed therebetween. The spacer member comprises an aperture that defines a growth compartment. At least one adhesive layer is adhered to the base or the coversheet in the growth compartment. A cold water-soluble gelling agent and an effective amount of a dry carbon dioxide-generating reagent are adhered to the at least one adhesive layer. The carbon dioxide-generating reagent consists essentially of particles having a diameter of less than 106 microns.

Abstract: Acoustic perfusion devices and processes for separating biomolecules from other material in a fluid mixture are disclosed. The devices include an inlet port, an outlet port, and a collection port that are connected to an acoustic chamber. An ultrasonic transducer creates an acoustic standing wave in the acoustic chamber that permits a continuous flow of fluid to be recovered through the collection port while keeping the biological cells within the acoustic chamber to be returned to the bioreactor from which the fluid mixture is being drawn.

Abstract: An object of the present invention is to purify and concentrate differentiating cells derived from ES cells, iPS cells, or the like without damaging them.

Abstract: A portable and mobile bioprocessing system and method for protein manufacturing that is compact, integrated and suited for on-demand production of any type of proteins and for delivery of the produced proteins to patients or for assay purposes. The portable system and method can also be used for efficient on-demand production of any type of protein with point-of-care delivery.

Abstract: The present invention relates to the field of stem cell biology, in particular the lineage specific differentiation of pluripotent or multipotent stem cells, which can include, but is not limited to, human embryonic stem cells (hESC), human induced pluripotent stem cells (hiPSC), somatic stem cells, cancer stem cells, or any other cell capable of lineage specific differentiation. Specifically described are methods to direct the lineage specific differentiation of hESC and/or hiPSC to nociceptors (i.e. nociceptor cells) using novel culture conditions. The nociceptors made using the methods of the present invention are further contemplated for various uses including, but limited to, use in in vitro drug discovery assays, pain research, and as a therapeutic to reverse disease of, or damage to, the peripheral nervous system (PNS). Further, compositions and methods are provided for producing melanocytes from human pluripotent stem cells for use in disease modeling.

Abstract: The present invention relates to a method for production of continuous cell lines comprising providing living cells of an animal or a human, irradiating said cells with UV light, proliferating said cells and selecting multiplying cells as cells of a continuous cell line.

Abstract: Provided herein are stimulated placental stem cells and methods of treating individuals having diseases or disorders of the circulatory system using stimulated placental cells. The invention also provides methods of inducing angiogenesis using such stimulated cells or populations of cells comprising such stimulated cells.

Abstract: The present invention is in the area of pluripotent stem cells and more particularly deals with a method to differentiate a vascular network from stem cells.

Abstract: The present invention provides means and methods for the generation of midbrain organoids which are useful for studying neurodevelopmental and neurodegenerative diseases. Neuroepithelial stem cells serve as a starting population for the generation of midbrain organoids by contacting them with differentiation medium under agitating conditions in three-dimensional cell culture comprising a matrix.

Abstract: The invention relates to culturing brain endothelial cells, and optionally astrocytes and neurons in a fluidic device under conditions whereby the cells mimic the structure and function of the blood brain barrier. Culture of such cells in a microfluidic device, whether alone or in combination with other cells, drives maturation and/or differentiation further than existing systems.

Abstract: The invention relates to culturing brain endothelial cells, and optionally astrocytes and neurons in a fluidic device under conditions whereby the cells mimic the structure and function of the blood brain barrier. Culture of such cells in a microfluidic device, whether alone or in combination with other cells, drives maturation and/or differentiation further than existing systems.

Abstract: Human pluripotent stem cells are differentiated in vitro into oligodendro-spheroids comprising oligodendrocytes for use in analysis, screening programs, and the like.

Abstract: The present invention relates to the use of warmers, or autonomous heat packs, for heating and maintaining a solution at a suitable temperature, for the period of time required to accomplish a chemical, biochemical or biological reaction, in particular in molecular biology or cell biology applications. Biology kits containing warmers are also part of this invention.

Abstract: Herein is reported a method for obtaining a B-cell comprising the following steps a) labeling B-cells, b) depositing the labeled B-cells as single cells, c) co-cultivating the single cell deposited B-cells with feeder cells, d) selecting a B-cell proliferating and secreting IgG in step c) and thereby obtaining a B-cell. The labeling can be of IgG+CD19+-B-cells, IgG+CD38+-B-cells, IgG+CD268+-B-cells, IgG?CD138+-B-cells, CD27+CD138+-B-cells or CD3?CD27+-B-cells. The method can comprise the step of incubating said B-cells at 37° C. for one hour in EL-4 B5 medium prior to the depositing step. The method can also comprise the step of centrifuging said single cell deposited B-cells prior to the co-cultivation. In the co-cultivation a feeder mix comprising interleukin-1beta, and tumor necrosis factor alpha and Staphylococcus aureus strain Cowans cells or BAFF or interleukin-2 and/or interleukin-10 and/or interleukin-6 and/or interleukin-4 can be used.

Abstract: The invention is directed to modified T cells, methods of making and using isolated, modified T cells, and methods of using these isolated, modified T cells to address diseases and disorders. In one embodiment, this invention broadly relates to TCR-deficient T cells, isolated populations thereof, and compositions comprising the same. In another embodiment of the invention, these TCR-deficient T cells are designed to express a functional non-TCR receptor. The invention also pertains to methods of making said TCR-deficient T cells, and methods of reducing or ameliorating, or preventing or treating, diseases and disorders using said TCR-deficient T cells, populations thereof, or compositions comprising the same.

Abstract: An object of the present invention is induction of CD4-positive T cells from pluripotent stem cells. This object is achieved by production of CD4-positive T cells by introducing a CD4 gene or a gene product thereof into T cells induced from pluripotent stem cells.

Abstract: This invention relates to the treatment of cancer in an individual by administration of a population of modified T cells that express a recombinant cAMP phosphodiesterase (PDE) or a fragment thereof and an antigen receptor which binds specifically to cancer cells in the individual. Populations of modified T cells and methods of producing populations of modified T cells are provided, along with pharmaceutical compositions and methods of treatment.

Abstract: The present invention relates to an apicidin composition for inducing differentiation of mesenchymal stem cells into cardiac-committed cells, and a method for inducing differentiation into cardiac-committed cells using the same. The present invention allows mesenchymal stem cells to be specifically induced to differentiate into cardiac-committed cells even when an apicidin is treated for only a very short period of time of 24 hours, thereby being capable of solving the extremely low cardiomyogenic differentiation efficiency of mesenchymal stem cells, high cost, and long-term problems in the conventional art, and thus the present invention can be usefully used for treating heart disease.

Abstract: Provided are methods for detecting and treating disease states, including cancer, diabetes, fibrosis, and autoimmune diseases, by detecting increased mechanical modulus, or stiffness, or targeting tissues having increased mechanical modulus, or stiffness. Practicing these methods provides specific and localized detection assays and therapies for these disease states. Provided are mechano-responsive cell systems that can selectively detect and treat cancer metastases by targeting the unique biophysical and mechanical properties in the tumor microenvironment. Provided are methods for making mechano-sensitive CAR T cells by using mechano-responsive promoter logic. Provided are blood tests using engineered stem cells that express reporters after the cells home to a specific niche and secrete the reporter into the blood, which can be then be detected with a blood test.

Abstract: Stromal stem cells are prospectively isolated from human bone marrow then expanded into clonal populations and cultured and used, the isolation being on the basis of expression of a cell surface marker, wherein the cell surface marker binds an antibody and wherein said antibody cross reacts with a cell surface marker found on mouse stromal stem cells or rat stromal stem cells, and optionally also on a cell of at least one other mammalian species selected from mouse, rat, horse, rabbit and pig cells. Useful stromal stem cell populations are positive for SDC2.

Abstract: The present invention provides a composition for isolating adipose-derived stromal cells, which comprises a Type I collagenase of 0.5-8% (v/v); a Trypsin of 0.1-0.6% (v/v); and a metal ion chelating agent of 0.01-0.2% (v/v). The present invention further provides a method for isolating adipose-derived stromal cells, which method comprises obtaining an adipose tissue; treating the adipose tissue with the composition of the present invention; centrifuging the adipose tissue; and isolating the adipose tissue to obtain the adipose-derived stromal cells. The present invention facilitates rapid isolation of mesenchymal stromal cells within a short period of time in operating rooms and can be applied to regenerative medicine in the future.

Abstract: The invention provides compositions, methods and systems for the ex vivo co-culture of renal cells, more specifically, for the co-culture of glomerulus-derived vascular endothelial cells and podocyte cells using apparatus that mimics the in vivo cellular architecture of the renal corpuscle. The invention described herein finds a variety of uses, for example, as a model system for the study of renal corpuscle function, including the filtration of the blood supply that occurs at the interface of the glomerulus and Bowman's capsule, normal physiology of those cell types, and as a model system for the study of renal disease, including the study of drug effects on the functioning of these cell types.

Abstract: The present invention relates to tunable hydrogels generated by selecting an amount of polymer and cross-linker such that the resulting hydrogel when covalently linked to an extracellular matrix (ECM) provides a biomimetic three-dimensional environment.

Abstract: The present invention provides methods of isolating a cancer-associated cell, such as a cancer stem cell or tumor initiating cell or a cell derived therefrom, from a mixture of cells, for example, a mixture of adherent cells in culture. Cell isolation is achieved by the application of selective detachment forces.

Abstract: An object of the present invention is to provide an agent for improving quality of an iPS cell, a method for producing an iPS cell, an iPS cell produced by such a method for production, and a composition for producing an iPS cell. The method for producing an iPS cell according to the present invention comprises the step of introducing (a) a nuclear reprogramming substance and (b) an H1foo gene or a gene product thereof into a somatic cell. High-quality iPS cells can be produced in greater quantity by introducing not only a nuclear reprogramming substance but also an H1foo gene or a gene product thereof into a somatic cell.

Abstract: The present disclosure provides methods for extracting viral particles from a sample comprising cells enclosing the viral particles. Also provided are kits that includes two or more items selected from a cell suspension solution, a cell lysis solution, a first fractionation solution, a second fractionation solution, a pellet suspension solution, a solid phase chromatography column, and a viral concentration set such as centrifugal filters.

Abstract: The present disclosure provides a method to enhance the production of virus in cultured fibroblasts by supplementing the cells with a TCA cycle intermediate, aketoglutarate, or a derivative thereof, wherein virus production is enhanced compared to the same method canied out in the absence of a-ketoglutarate or the derivative thereof. In view of the art, the method provides an unexpected improvement on methods routinely practiced.

Abstract: In some aspects, isolated transgenic cells (e.g., transgenic T cells) are provided that comprise or express a transgene and DHFRFS and/or TYMSSS. Methods for selecting transgenic cells are also provided.

Abstract: The present invention relates to methods and compositions for improving the productivity of recombinant vitamin K dependent protein expression in host cells.

Abstract: A composition comprised of a perodixase, a catalase and a peroxide that, in combination, is suitable for degradation of pollen wall sporopollenin, and coatings comprising sporopollenin in a matrix that can be selectively degraded through application of a peroxidase, a catalase and a peroxide to the sporopollenin matrix.

Abstract: The invention relates to a polypeptide for the enzymatic detoxification of zearalenone, said polypeptide being a monooxygenase which converts the keto group in position 7 of zearalenone into an ester group, the monooxygenase in particular being an amino acid sequence selected from the group comprising sequence ID No. 1, 2 and 3 or a functional variant thereof.

Abstract: The invention is directed to p-coumaroyl-CoA:monolignol transferase enzymes, nucleic acids encoding p-coumaroyl-CoA:monolignol transferase enzymes, and inhibitory nucleic acids adapted to inhibit the expression and/or translation of p-coumaroyl-CoA:monolignol transferase RNA; expression cassettes, plant cells, and plants that have or encode such nucleic acids and enzymes; and methods of making and using such nucleic acids, enzymes, expression cassettes, cells, and plants.

Abstract: The invention relates to transaminases that are particularly useful for catalyzing the conversion of amine substrates to ketone products and/or vice versa.

Abstract: The present disclosure provides engineered transaminase enzymes having improved properties as compared to a naturally occurring wild-type transaminase enzyme. Also provided are polynucleotides encoding the engineered transaminase enzymes, host cells capable of expressing the engineered transaminase enzymes, and methods of using the engineered transaminase enzymes to synthesize a variety of chiral compounds.

Abstract: Presented herein are transposase enzymes and reaction conditions for improved fragmentation and tagging of nucleic acid samples, in particular altered transposases and reaction conditions which exhibit improved insertion sequence bias, as well as methods and kits using the same.

Abstract: A DNA polymerase, having an amino acid sequence represented by SEQ ID NO:2, or a derivative of the amino acid sequence by substitution, deletion, or addition of at least one amino acid residue. The DNA polymerase is a hybrid DNA polymerase prepared by inserting a thioredoxin binding domain (TBD) of bacteriophage T7 DNA polymerase into a DNA polymerase I (Sau) of Staphylococcus aureus. A method for preparing the DNA polymerase includes: 1) determining a corresponding position and a target substitution sequence in Sau protein for the TBD of the bacteriophage T7 DNA polymerase; 2) devising and synthesizing a primer according to a gene sequence of Sau and a sequence TBD published by GenBank; 3) cloning the Sau-TBD segment acquired in (2) to an expression vector pTrc99A to construct a recombinant vector pTrc99A-Sau-TBD; and 4) transforming Escherichia coli by the recombinant vector pTrc99A-Sau-TBD and inducing protein expression.

Abstract: The invention relates to modified polymerase enzymes which exhibit improved incorporation of nucleotide analogues bearing substituents at the 3? position of the sugar moiety that are larger in size than the naturally occurring 3? hydroxyl group. Also described are methods of using the polymerases to incorporate nucleotides into polynucleotides, particularly in the context of DNA sequencing.

Abstract: A secretion signal peptide sequence (SP) in combination with a cleavage inhibition sequence (CIS) fused to a structural gene sequence in a recombinant expression system can be used to express a full length protein with an SP in a cell. Such a fusion protein may be purified to homogeneity from a membrane fraction of the cell. The SP in combination with the CIS is a protein transduction domain that exhibits superior intracellular protein transduction efficiency when the SP precedes the CIS in a N to C-terminus direction.

Abstract: The present disclosure provides variant Cas9 proteins, nucleic acids encoding the variant Cas9 proteins, and host cells comprising the nucleic acids. The present disclosure provides systems that include a subject variant Cas9 protein (and/or a nucleic acid encoding the variant Cas9 protein) and a Cas9 guide RNA. In some cases, a subject system includes a PAMmer and/or a donor polynucleotide. The variant Cas9 proteins and the nucleic acids encoding the variant Cas9 proteins are useful in a wide variety of methods, which are also provided. In some embodiments, a variant Cas9 protein includes a RuvC domain, an HNH domain, and a disrupted RuvC/HNH linker region that reduces the RuvC cleavage activity of the protein. In some embodiments, a variant Cas9 protein includes a deletion (of all or a part of the HNH domain) or an insertion (within the HNH domain) that reduces the HNH cleavage activity of the protein.

Abstract: A mutant of Endos2 includes one or more mutations in the sequence of a wild-type EndoS2 (SEQ ID NO:1), wherein the one or more mutations are in a peptide region located within residues 133-143, residues 177-182, residues 184-189, residues 221-231, and/or residues 227-237, wherein the mutant of EndoS2 has a low hydrolyzing activity and a high tranglycosylation activity, as compared to those of the wild-type EndoS2. A method for preparing an engineered glycoprotein using the mutant of EndoS2 includes coupling an activated oligosaccharide to a glycoprotein acceptor. The activated oligosaccharide is a glycan oxazoline.

Abstract: The present invention relates to isolated polypeptides having cellulolytic enhancing activity and polynucleotides encoding the polypeptides. The invention also relates to nucleic acid constructs, vectors, and host cells comprising the polynucleotides as well as methods of producing and using the polypeptides.

Abstract: The invention relates to a host cell comprising at least four different heterologous polynucleotides chose from the group of polynucleotides encoding cellulases, hemicellulases and pectinases, wherein the host cell is capable of producing the at least four different enzymes chosen from the group of cellulases, hemicellulases and pectinases, wherein the host cell is a filam-tous fungus and is capable of secretion of the at least four different enzymes. This host cell can suitably be used for the production of an enzyme composition that can be use in a process for the saccharification of cellulosic material.

Abstract: The present invention relates to pullulanase variants comprising substitutions of the parent pullulanase at one or more positions corresponding to positions 393, 143, 150, 243, 244, 345, 346, 368, 370, 373, 381, 382, 385, 387, 402, 429, 430, 431, 432, 456, 486, 492, 610, 624, 631, 632, 665 and 699 of the polypeptide of SEQ ID NO: 3. The present invention also relates to polynucleotides encoding the variants; nucleic acid constructs, vectors, and host cells comprising the polynucleotides; and methods of using the variants.

Abstract: The present invention relates to isolated polypeptides having lysozyme activity and polynucleotides encoding the polypeptides. The invention also relates to nucleic acid constructs, vectors, and host cells comprising the polynucleotides as well as methods of producing and using the polypeptides.