Abstract: The present invention provides peptide-ligand conjugates containing a peptide, where a free form of the peptide is capable of displacing a second ligand specific for the peptide. The invention also provides a method of selecting a cell. The method includes the steps of contacting a cell population with a peptide-ligand conjugate, wherein the conjugate comprises a first ligand specific for a cell in the population conjugated to a peptide capable of displacing a second ligand specific for the peptide, thereby forming a cell-conjugate complex; contacting the peptide-ligand conjugate with the second ligand; and isolating the cell-conjugate complex. The method can further include the step of adding a free form of the peptide to release the cell from the complex. The invention additionally provides a method of diagnosing a condition using peptide-ligand conjugates.

Abstract: In accordance with the present invention, provided is a method for producing human circulating dendritic cells (cirDC) for therapeutic use, by depleting a human blood leukocyte composition of B cells, T cells and monocytes. Also provided are compositions containing cirDC for therapeutic use.

Abstract: The present invention provides peptide-ligand conjugates containing a peptide, where a free form of the peptide is capable of displacing a second ligand specific for the peptide. The invention also provides a method of selecting a cell. The method includes the steps of contacting a cell population with a peptide-ligand conjugate, wherein the conjugate comprises a first ligand specific for a cell in the population conjugated to a peptide capable of displacing a second ligand specific for the peptide, thereby forming a cell-conjugate complex; contacting the peptide-ligand conjugate with the second ligand; and isolating the cell-conjugate complex. The method can further include the step of adding a free form of the peptide to release the cell from the complex. The invention also provides a subpopulation of cells containing two or more specifically isolated cells and methods of isolating cell subpopulations. The invention additionally provides a method of diagnosing a condition using peptide-ligand conjugates.

Abstract: A device and method of use are disclosed to facilitate fluid transfers during sterile closed system processing procedures. The device is configured to create connections for the transfer of fluid between various components, along a closed sterile fluid passageway and, eventually, either to a fluid container or to a patient. As a result, the device and method of use of the present invention reduce the risk of contamination to the fluids, such as reagents, medicaments and cellular products, and increase user or technician safety during processing procedures.

Abstract: A device and method of use are disclosed to facilitate fluid transfers during sterile closed system processing procedures. The device is configured to create connections for the transfer of fluid between various components, along a closed sterile fluid passageway and, eventually, either to a fluid container or to a patient. As a result, the device and method of use of the present invention reduce the risk of contamination to the fluids, such as reagents, medicaments and cellular products, and increase user or technician safety during processing procedures.

Abstract: An apparatus and method of use are disclosed to automatically process and transfer fluids. The apparatus may include a user-interface, clamps, pumps, spinning membranes, pressure transducers, fluid detectors, solution towers, weight scales and tubing sets to control and monitor cell washing, concentrating and harvesting procedures. In addition, the apparatus may also be used to control and monitor a variety of fluid transfer procedures. The apparatus may also be used to execute default or user-defined cell washing and fluid transfer procedures.

Abstract: A method for producing human dendritic cells for therapeutic purposes which allows culture-deriving dendritic cells using no cytokines, or reduced cytokines. The method involves culturing mononuclear cells from blood or bone marrow in a medium containing at least one agent such as a calcium ionophore, e.g. A23187, theophylline, protaglandin E1, dibutyryl cyclic AMP, Vitamin D3, Vitamin E, retinoic acid, or a fatty acid. The culture is maintained for a sufficient time, typically 4-14 days, to produce a culture enriched for dendritic cells, as evidenced by at least about 2.5% of total cells exhibiting dendritic cell processes, or a dendritic dell antigen such as CD80, CD86, or CD1a. Also provided is a method to produce antigen-specific human T-cells by pulsing the dendritic cells obtained by the method of the invention with an antigen such as a viral, tumor, bacterial, or cell surface antigen, and then co-culturing T-cells with the antigen-pulsed dendritic cells.

Abstract: A bag or reservoir for recirculation washing of blood cells having a top outlet port and bottom inlet port. A method of recirculation washing of blood cells using the bag in conjunction with a spinning membrane filter. The method can be used in an instrument for magnetic cell selection or a stand-alone cell washing apparatus.

Abstract: A bag or reservoir for recirculation washing of blood cells has a top outlet port and bottom inlet port. A method of recirculation washing of blood cells uses the bag in conjunction with a spinning membrane filter. The method can be used in an instrument for magnetic cell selection or a stand-alone cell washing apparatus.

Abstract: A composition comprising genetically altered human neutrophil precursor cells, wherein the cellular component is comprised of at least about 16% human myeloblasts and promyelocytes, which have been derived from neutrophil progenitor cells obtained from peripheral blood, bone marrow or cord blood, and less than about 5% colony forming units (CFU) of at least about 50 cells is provided. An alternative composition comprising genetically altered human neutrophil precursor cells, wherein the cellular component is comprised of at least about 16% CD15+CD11b- cells and less than about 5% colony forming units (CFU) of at least about 50 cells also is provided, wherein at least about 60% of the CD15+CD11b- cells are myeloblasts and promyelocytes.

Abstract: Provided are serum-free, animal protein-free media formulations to be used in conjunction with hematopoietic growth factors for the in vitro growth of human neutrophil and megakaryocyte precursors. The medium contains a base medium, corticosteroid, transferrin, insulin, cholesterol, ethanolamine, and human albumin. Also provided are methods for preparing serum-free, animal protein-free suspensions of human hematopoietic precursor cells wherein the cellular component contains at least about 16% neutrophil precursors and at least about 1% megakaryocyte precursors. Serum-free, animal protein-free suspensions of human hematopoietic cells are provided wherein the cellular component comprises at least about 30%, preferably greater than 60% neutrophil precursors. The neutrophil precursors are comprised of blast cells, promyclocytes, neutrophilic myelocytes, and neutrophilic metamyelocytes.

Abstract: The invention provides a nonenzymic method for the release of cells which have been selected from a heterogeneous cell suspension by antibody-mediated binding to beads or other solid support. The method entails forming within the cell suspension a complex comprising the solid support linked to a primary monoclonal antibody, which in turn is bound to a cell surface antigen on the target cells. The complex is separated from the cell suspension, and then contacted with a specific peptide which binds to the primary antibody, displacing the antibody from the cell surface antigen, thereby releasing the target cell from the complex. The invention also provides methods for positive/negative cell selection wherein target cells having a first antigen are selected from a heterogeneous cell suspension containing undesired cells having a second antigen.

Abstract: The invention provides a method of treating a patient having a reduced population of neutrophils following a myeloablative cancer treatment such as high dose chemotherapy. Following myeloablative therapy, a cell composition of at least 25% neutrophil precursors, i.e. promyelocytes, myelocytes, and metamyelocytes, is administered to the patient. Thereafter, the neutrophil precursors differentiate rapidly in vivo to replenish the supply of mature neutrophils for fighting infection. The method is used to reduce the neutropenic window between the time of myeloablative therapy and the time required for infused stem cells to proliferate and differentiate into mature neutrophils.

Abstract: The invention provides a non-enzymatic method for the release of cells which have been positively selected from a heterogeneous cell suspension by antibody-mediated binding to beads or other solid support. The method entails forming within the cell suspension a complex comprising the solid support linked to a primary antibody, which in turn is specifically bound to a cell surface antigen on the target cells. The complex is separated from the cell suspension, and then contacted with a specific peptide which binds to the primary antibody, displacing the antibody from the cell surface antigen, thereby releasing the target cell from the complex. The invention also provides methods for positive/negative cell selection wherein target cells having a first antigen are selected from a heterogeneous cell suspension containing undesired cells having a second antigen.

Abstract: The present invention provides a composition of human neutrophil precursor cells having at least 37% myeloblasts and promyeloblasts. Also provided are hematopoietic cell suspensions that have human neutrophil precursor cells and at least one hematopoietic growth factor.

Abstract: A suspension comprising human neutrophil precursor cells, wherein the cellular component is comprised of at least about 16% human myeloblasts and promyeclocytes, which have been derived from neutrophis progenitor cells obtained from peripheral blood, bone marrow or cord blood, and less than about 5% colony forming units (CFU) of at least about 50 cells is provided. An alternative suspension comprising human neutrophil precursor cells, wherein the cellular component is comprised of at least about 16% CD15+CD11b- cells and less than about 5% colony forming units (CFU) of at least about 50 cells also is provided, wherein at least about 60% of the CD15+CD11b- cells are myeloblasts and promyelocytes. The suspensions of the invention are useful in methods for increasing neutrophil populations in a patient having a reduced populations of neutrophils.

Abstract: The invention provides a method of treating a patient having a reduced population of neutrophils following a myeloablative cancer treatment such as high dose chemotherapy. Following myeloablative therapy, a cell composition of at least 25% neutrophil precursors, i.e. promyelocytes, myelocytes, and metamyelocytes, is administered to the patient. Thereafter, the neutrophil precursors differentiate rapidly in vivo to replenish the supply of mature neutrophils for fighting infection. The method is used to reduce the neutropenic window between the time of myeloablative therapy and the time required for infused stem cells to proliferate and differentiate into mature neutrophils.