Production of megakaryocytes by the use of human mesenchymal stem cells

• US 20010005591A1
• Filed: 02/09/2001
• Published: 06/28/2001
• Est. Priority Date: 05/22/1998
• Status: Active Grant

• Current Assignee: Mesoblast International Srl
• Original Assignee: Osiris Therapeutics Incorporated (Smith & Nephew PLC)
• Inventors: Thiede, Mark A., Qasba, Pankaj
• Granted Patent: US 7,153,500 B2
• Time in Patent Office: Days
• US Class Current: 435/373
• CPC Class Codes: A61K 2035/124 the cells being hematopoietic, bone marrow derived or blood cells
• CPC Class Codes: C12N 2501/125 Stem cell factor [SCF], c-kit ligand [KL]
• CPC Class Codes: C12N 2501/145 Thrombopoietin [TPO]
• CPC Class Codes: C12N 2501/22 Colony stimulating factors (G-CSF, GM-CSF)
• CPC Class Codes: C12N 2501/23 Interleukins [IL]
• CPC Class Codes: C12N 2501/26 Flt-3 ligand (CD135L, flk-2 ligand)
• CPC Class Codes: C12N 2502/1358 Bone marrow mesenchymal stem cells (BM-MSC)
• CPC Class Codes: C12N 2510/00 Genetically modified cells
• CPC Class Codes: C12N 5/0644 Platelets; Megakaryocytes

Abstract

The present invention is directed to hematopoietic progenitor cells isolated from a tissue specimen, such as marrow cells or peripheral blood, and to the method of co-culturing isolated hematopoietic progenitor cells with human mesenchymal stem cells to induce megakaryocyte differentiation and platelet production. In addition, hematopoietic stem cells can be genetically engineered to carry genes of interest particularly for the expression of physiologically active proteins. In the presence of mesenchymal stem cells, the transduced cells carry the new genetic material and express gene products that can be used to modulate blood disorders.

Claims

1. A method of producing megakaryocytes in vitro comprising co-culturing human mesenchymal stem cells with CD34+ cells to induce the CD34+ cells to differentiate into megakaryocytes.

2. The method of

claim 1

wherein the mesenchymal stem cell population is allogeneic or autologous to the hematopoietic progenitor cell population.

3. The method of

claim 1

wherein the cells are in close physical association in a monolayer.

4. A method for treating a patient in need of megakaryocytes, comprising administering to the patient human mesenchymal stem cells in an amount effective to produce megakaryocytes.

5. A method for treating a patient in need of megakaryocytes comprising administering to the patient mesenchymal stem cells and CD34+ cells in an amount effective to produce megakaryocytes.

6. A method of producing genetically modified megakaryocytes, comprising transducing hematopoietic progenitor cells with exogenous genetic material;

and culturing the transduced hematopoietic cells in the presence of mesenchymal stem cells to induce differentiation of the transduced hematopoietic cells into megakaryocytes which contain the exogenous genetic material.

7. A composition of genetically modified megakaryocytes.

8. A composition of genetically modified platelets.