Maintenance and propagation of mesenchymal stem cells

• US 8,388,947 B2
• Filed: 11/16/2011
• Issued: 03/05/2013
• Est. Priority Date: 01/22/2007
• Status: Active Grant

• Current Assignee: US Department of Veterans Affairs (Government of the United States of America), The Board of Trustees of the University of Arkansas (University of Arkansas System)
• Original Assignee: US Department of Veterans Affairs (Government of the United States of America), The Board of Trustees of the University of Arkansas (University of Arkansas System)
• Inventors: Chen, Xiao-Dong, Jilka, Robert L.
• Primary Examiner(s): Long, Scott
• Application Number:
  US13/297,966
• Publication Number:
  US 20120064046A1
• Time in Patent Office: 475 Days
• Field of Search: None
• US Class Current: 424/93.7
• CPC Class Codes: A61K 2035/124 the cells being hematopoietic, bone marrow derived or blood cells
• CPC Class Codes: A61K 35/28 Bone marrow; Haematopoietic stem cells; Mesenchymal stem cells of any origin, e.g. adipose-derived stem cells
• CPC Class Codes: A61P 19/08 for bone diseases, e.g. rachitism, Paget's disease
• CPC Class Codes: C12M 21/08 for producing artificial tissue or for ex-vivo cultivation of tissue prostheses A61F2/00, grafts A61L27/00
• CPC Class Codes: C12M 25/14 Scaffolds; Matrices in general C12N5/0068
• CPC Class Codes: C12N 2513/00 3D culture
• CPC Class Codes: C12N 2533/90 Substrates of biological origin, e.g. extracellular matrix, decellularised tissue
• CPC Class Codes: C12N 5/0662 Stem cells
• CPC Class Codes: C12N 5/0663 Bone marrow mesenchymal stem cells (BM-MSC)
• CPC Class Codes: C12N 5/0668 Mesenchymal stem cells from other natural sources

Abstract

Various embodiments of the present invention include compositions, materials and methods for maintaining and propagating mammalian mesenchymal stem cells in an undifferentiated state in the absence of feeder cells and applications of the same.

Claims

1. A composition for maintaining mammalian mesenchymal stem cells in culture in an undifferentiated state comprising an in vitro extracellular matrix, wherein the extracellular matrix restrains differentiation of the mammalian mesenchymal stem cells, and wherein said extracellular matrix comprises type I collagen, type III collagen, type V collagen, syndecan-1, fibronectin, decorin, biglycan, perlecan, and laminin.

2. The composition of claim 1 wherein said composition is essentially free of feeder cells.

3. The composition of claim 1 wherein said mammalian mesenchymal stem cells are selected from the group consisting of human mesenchymal stem cells and murine mesenchymal stem cells.

4. The composition of claim 1 wherein said extracellular matrix is a marrow stromal cell derived extracellular matrix.

5. The composition of claim 1 wherein said extracellular matrix is a three-dimensional matrix.

6. The composition of claim 1 wherein said extracellular matrix is manufactured by culturing marrow stromal cells, lysing the marrow stromal cells and removing the lysed marrow stromal cells by washing.

7. The composition of claim 1, wherein the extracellular matrix comprises an inhibitor of BMP2 activity.