Nexell Therapeutics Inc.

Baxter International Inc.

Baxter Healthcare Inc.

Dennis E Van Epps

Stephen L Smith

James G Bender

Kristen L Unverzagt

Phillip B Maples

Maureen F Loudovaris

Susan M Maciukas

Xiaoying Qiao

Daniel F Bischof

John R Chapman

Richard C Giesler

Dale R Ellis

Jeffrey A Martinson

William C Lake

Frederick Aono

Frederick M Aono

Mortimer T Alzona

Mona Vachula

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. Useful for treatment of viral or bacterial infections, useful as a cancer vaccine, useful to induce tolerance of allo- or xeno- graft.

Cytokine-free culture of dendritic cells

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.

In vitro-derived genetically altered human neutrophil precursor cells

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. Serum-free, animal protein-free cells suspensions are provided wherein the cellular component comprises at least about 3%, preferably greater than 8% megakaryocyte precursors. Also provided are serum-free, animal protein free cell suspensions wherein the cellular component comprises colony-forming cells and cluster-forming cells.

Preparation of serum-free suspensions of human hematopoietic cells or

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.

Infusion of neutrophil precursors for treatment of neutropenia

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.

In-vitro-derived human neutrophil precursor cells

A method combining the techniques of immunoaffinity separation and continuous flow centrifugal separation is provided for selective separation of a nucleated heterogeneous cell population from a heterogeneous cell mixture. The heterogeneous cell mixture is intimately contacted to promote binding thereto by particles having attached a substance that actively binds to a specific desired type of cell out of the cell mixture. The particles are selected so that the sedimentation velocity of the particle/cell conjugate differs sufficiently from those of other cells in the cell mixture to allow its separation by means of a continuous flow cell separator. The method rapidly processes large volumes of cell mixture with the high accuracy expected of immunoaffinity separation and can be used to separate, for example, various types of leukocytes from whole blood, bone marrow concentrate, or a peripheral blood stem cell concentrate; or precursors of lymphokine activated killer cells, tumor infiltrating lymphocyte cells, or activated killer monocytes from lymphocyte or monocyte cell concentrates or from a tissue cell preparation.

Continuous process for the separation of biologic components from

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.

Method for increasing neutrophil populations using in vitro-derived

A composition of human neutrophil precursor cells is disclosed wherein at least 16% of the cells are human myeloblasts and promyelocytes. The myeloblasts and promyelocytes are derived from human neutrophil progenitor cells that were obtained from peripheral blood, bone marrow or cord blood. The neutrophil precursor cells contain less than 5% colony forming units. Also disclosed are human neutrophil precursor cells made up of about 16% CD15+CD11b- cells and less than 5% colony forming units and methods of preparing these compositions.

Growing community of inventors

Cary, IL, United States of America

Dennis E Van Epps