Abstract: The present invention relates to methods for separating target cells from non-target cells using immunorosettes and magnetic particles. The method involves contacting a sample containing target cells and secondary targets such as erythrocytes with an antibody composition which allows immunorosettes of the target cells and the secondary targets to form. The sample is subsequently contacted with a second antibody composition which allows the binding of magnetic particles to the formed immunorosettes and free secondary targets. The immunorosettes and secondary targets labeled with magnetic particles are separated from non-target cells using a magnetic field. The antibody composition optionally contains bifunctional antibodies or tetrameric antibody complexes.

Abstract: An improved method for the preparation of tetrameric antibody complexes directly on the surface of target entities in a sample is described. In particular, this method involves linking, in the sample, a first target entity with a second target entity in a sample using antibodies with specificity for the first and second target entities.

Abstract: This disclosure provides compositions and methods for the activation and expansion of human T cells or NK cells using soluble monospecific antibody complexes.

Abstract: The present disclosure provides methods of generating germ layers from stem cells comprising culturing the stem cells in a culture medium having an osmolality less than 340 mOsm/kg. The present disclosure also includes a method to generate different cell lineages from the germ layers as well as to detect them by immunological methods. The present disclosure further provides methods for the generation, isolation, cultivation and propagation of committed progenitor cells and for the production of differentiated cells from the three germ layers. The present disclosure also provides culture media for use in inducing the three germ layers.

Abstract: Surfaces contactable by a bulk liquid for use in particle isolation are described. The surfaces comprise a plurality of ribs spaced apart to capillarily retain a portion of the bulk liquid and particles of interest therebetween. Collectively, the portion of the bulk liquid retained by the plurality of ribs on the surface forms a liquid film. The one or more particles received within the space and enveloped by the liquid film may be protected from one or more forces exerted by a draining meniscus passing over the surface.

Abstract: A meniscus reducing member for use in a vessel for containing a liquid may include a physical surface feature overlying at least a portion of an interior surface of the vessel. The physical surface feature may have first and second inner surfaces that are generally parallel and at least a third surface extending between the first and second surfaces. The first inner surface, second inner surface and third surfaces may be configured to physically alter a receding contact angle between the liquid and the physical surface feature. A coating material may be applied to at least one of the surfaces of the physical surface feature to chemically alter the receding contact angle between the liquid and the coated surface whereby the receding contact angle formed between the liquid and the meniscus reducing member is between about 90 degrees and less than 180 degrees.

Abstract: The present invention relates to methods for separating target cells from non-target cells using immunorosettes and magnetic particles. The method involves contacting a sample containing target cells and secondary targets such as erythrocytes with an antibody composition which allows immunorosettes of the target cells and the secondary targets to form. The sample is subsequently contacted with a second antibody composition which allows the binding of magnetic particles to the formed immunorosettes and free secondary targets. The immunorosettes and secondary targets labeled with magnetic particles are separated from non-target cells using a magnetic field. The antibody composition optionally contains bifunctional antibodies or tetrameric antibody complexes.

Abstract: The disclosure provides a method of culturing cells of the mesenchymal cell lineage, said method comprising contacting the cells with a culture media comprising a CSF1R kinase inhibitor. The disclosure also provides a method of culturing cells from bone marrow and/or compact bone to enrich the cells with cells of the mesenchymal cell lineage comprising contacting the cells with a culture media comprising a CSF1R kinase inhibitor. Cell culture media comprising a CSF1R kinase inhibitor and useful for culturing cells of the mesenchymal cell lineage and/or enriching cells of the mesenchymal cell lineage is also provided.

Abstract: An improved method for the preparation of target specific surfaces and uses thereof is described. In particular, the surfaces are bound to mono-specific tetrameric antibody complexes prior to their addition to a sample containing target entities and separating them from non-target entities.

Abstract: A meniscus reducing member for use in a vessel for containing a liquid may include a physical surface feature overlying at least a portion of an interior surface of the vessel. The physical surface feature may have first and second inner surfaces that are generally parallel and at least a third surface extending between the first and second surfaces. The first inner surface, second inner surface and third surfaces may be configured to physically alter a receding contact angle between the liquid and the physical surface feature. A coating material may be applied to at least one of the surfaces of the physical surface feature to chemically alter the receding contact angle between the liquid and the coated surface whereby the receding contact angle formed between the liquid and the meniscus reducing member is between about 75 degrees and 110 degrees.

Abstract: The present disclosure provides methods of generating germ layers from stem cells comprising culturing the stem cells in a culture media having osmolality ranges that promote the generation of specific germ layer progenitor cells. The present disclosure also includes a method to generate different cell lineages from the germ layers as well as to detect them by immunological methods. The present disclosure further provides methods for the generation, isolation, cultivation and propagation of committed progenitor cells and for the production of differentiated cells from the three germ layers. The present disclosure also provides culture media and kits for use in inducing the three germ layers.

Abstract: A magnetic particle comprises a polysaccharide matrix and a plurality of magnetic crystals dispersed in the matrix. A method for making magnetic particles comprises combining a basic solution with a metal ion solution and allowing the metal ions to oxidize to form magnetic crystals, and combining the magnetic crystals with a polysaccharide solution to form the magnetic particles.

Abstract: An insert for a centrifuge tube suitable for use in density gradient separation is described. The insert includes a member sized to fit within the tube for dividing the tube into a top portion and a bottom portion. Optionally the insert has a support extending or depending from the member for positioning the member within the tube. At least two openings are located on the member so that a first opening is closer to a bottom end of the tube relative to a second opening when the insert is positioned in the centrifuge tube. Also described are methods for separating a target population of cells from a sample using the insert for a centrifuge tube.

Abstract: A filter apparatus comprises a filter having first and second opposed surfaces. A first reservoir is positioned adjacent with the first surface and in communication with the first surface, and a first inlet-outlet port is in communication with the first reservoir and spaced from the first surface. A second reservoir is positioned adjacent the second surface, and in communication with the second surface, and a second inlet-outlet port is in communication with the second reservoir and spaced from the second surface. A filter plate system comprises a reservoir plate. The reservoir plate comprises at least one reservoir well. The filter plate system further comprises a strainer plate. The strainer plate comprises at least one strainer well, which is removably receivable in the reservoir well. The strainer well comprises at least one mesh wall portion.

Abstract: The present disclosure provides a method of determining the frequency and proliferative potential of progenitor cells in a starting cell preparation comprising (a) culturing the cell preparation for 5-10 days in a culture medium comprising FLT-3 Ligand (FL) and Thrombopoietin (TPO); (b) detecting and enumerating the colonies; and (c) assessing the frequency and proliferative potential of progenitor cells in the starting cell preparation based on the number of colonies in step (b). The number of colonies in step (b) correlates to the number of colonies in a standard 14-day colony-forming cell assay. The method is particularly useful for predicting and evaluating the suitability of grafts for transplantation.

Abstract: A cell culture device comprises a well. A plurality of microwells are within the well, and a first common fluid volume is within the well above the microwells. A set of sub-microwells are within each microwell, and a second common fluid volume is within each microwell above the set of sub-microwells.

Abstract: A meniscus reducing member for use in a vessel for containing a liquid including a surface feature overlying at least a portion of an interior surface of the vessel. The surface feature includes at least two surfaces for contacting the liquid that cooperate to reduce a width of a meniscus formed at an interface between the liquid and the surface feature by physically altering a contact angle between the liquid and the surface feature.

Abstract: The disclosure provides a method of culturing cells of the mesenchymal cell lineage, said method comprising contacting the cells with a culture media comprising a CSF1R kinase inhibitor. The disclosure also provides a method of culturing cells from bone marrow and/or compact bone to enrich the cells with cells of the mesenchymal cell lineage comprising contacting the cells with a culture media comprising a CSF1R kinase inhibitor. Cell culture media comprising a CSF1R kinase inhibitor and useful for culturing cells of the mesenchymal cell lineage and/or enriching cells of the mesenchymal cell lineage is also provided.

Abstract: A meniscus reducing member for use in a vessel for containing a liquid may include a physical surface feature overlying at least a portion of an interior surface of the vessel. The physical surface feature may have first and second inner surfaces that are generally parallel and at least a third surface extending between the first and second surfaces. The first inner surface, second inner surface and third surfaces may be configured to physically alter a receding contact angle between the liquid and the physical surface feature. A coating material may be applied to at least one of the surfaces of the physical surface feature to chemically alter the receding contact angle between the liquid and the coated surface whereby the receding contact angle formed between the liquid and the meniscus reducing member is between about 75 degrees and 110 degrees.

Abstract: The present application is directed to methods of improving cell culture vessel assays. In one aspect the application is directed to a method of reducing the curvature of the meniscus comprising applying a coating material to the interior wall of the vessel, wherein the coating material provides a receding contact angle of about 90 degrees with aqueous solutions and culture media. In another aspect, the application is directed to a method of labeling cells in a first solution by generating droplets of a second solution containing at least one cell-labelling agent and allowing the droplets of the second solution to contact the surface of the first solution.