Maintenance and propagation of mesenchymal stem cells

• US 10,676,718 B2
• Filed: 01/11/2018
• Issued: 06/09/2020
• 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: 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:
  US15/868,721
• Publication Number:
  US 20180127720A1
• Time in Patent Office: 880 Days
• 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 method of generating bone in a patient in need thereof comprising:

(i) obtaining mammalian mesenchymal stem cells;

(ii) producing a bone forming composition by culturing said mammalian mesenchymal stem cells in vitro on a three-dimensional marrow stromal cell derived extracellular matrix, isolating undifferentiated mammalian mesenchymal stem cells from said culture, and combining said undifferentiated mammalian mesenchymal stem cells with a transplantation vehicle; and

(iii) administering said bone forming composition to said patient;

wherein said extracellular matrix comprises type I collagen, type III collagen, type V collagen, syndecan-1, fibronectin, decorin, biglycan, perlecan, and laminin.

2. The method 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.

3. The method of claim 1, wherein said extracellular matrix is manufactured by culturing marrow stromal cells on a substrate, lysing the marrow stromal cells, and removing the lysed marrow stromal cells by washing.

4. The method of claim 1, wherein said mammalian mesenchymal stem cells are autologous to said patient.

5. The method of claim 1, wherein said patient is a human patient.

6. The method of claim 5, wherein said human patient is an aging human patient.

7. The method of claim 1, wherein said administering of step (iii) comprises local administration or systemic administration.