# US 2010/0167316 A1 · Enrichment of Tissue-Derived Adult Stem Cells Based on Retained Extracellular Matrix Material > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 2010/0167316 A1 - Document ID: US20100167316A1 - Status: Published application - Kind: A1 - Application number: 12/594,493 - Filed: 2008-04-03 - Published: 2010-07-01 - Earliest claimed: 2007-04-03 - Assignees: The Cleveland Clinic Foundation - Inventors: George Muschler; Tonya Caralla; Vincent Hascall; Ronald Midura - Web page: https://patentgrep.com/patent/US20100167316A1 ## CPC - C12N 5/0663: {Bone marrow mesenchymal stem cells (BM-MSC)} - C12N 2500/38: Vitamins - C12N 2501/39: Steroid hormones - C12N 2501/905: Hyaluronic acid - C12N 2533/80: Hyaluronan - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) ## Abstract Methods for enriching, detecting, or using adult stem cells through the use of recognition ligands that specifically bind to ECM components retained to the surfaces of adult stem cells are described. An ECM component such as hyaluronan that is retained to the surfaces of adult stem cells when removed from animal tissues can be used to detect a diverse population of adult stem cells based on the nature of the ECM niche region in which the adult stem cells normally reside. For example, a separation method such as magnetic separation can be used to detect and isolate or enrich adult stem cells based on a recognition ligand that is specific for an ECM component that is retained to the surfaces of adult stem cells to a greater degree than to other cells in the population. ## Claims ### Claim 1 A method for enriching adult stem cells comprising: obtaining a population of cells including one or more adult stem cells from animal tissue; contacting the population of cells with a recognition ligand specific for an ECM component retained to the surfaces of adult stem cells in said population; and using a separation method to remove cells from said population that are not bound to the recognition ligand, thereby enriching adult stem cells that are bound to the recognition ligand via said retained ECM component. ### Claim 2 (depends on 1) The method of claim 1, wherein the animal tissue is connective tissue. ### Claim 3 (depends on 1) The method of claim 1, wherein the separation method comprises magnetic separation. ### Claim 4 (depends on 3) The method of claim 3, wherein the separation method further comprises selective retention using a porous matrix. ### Claim 5 (depends on 1) The method of claim 1, wherein the separation method comprises an affinity column method. ### Claim 6 (depends on 1) The method of claim 1, wherein the retained ECM component is hyaluronan. ### Claim 7 (depends on 6) The method of claim 6, wherein the recognition ligand is hyaluronan binding protein. ### Claim 8 (depends on 1) The method of claim 1, wherein the adult stem cells comprise connective tissue progenitor cells. ### Claim 9 (depends on 1) The method of claim 1, wherein the method increases the prevalence of adult stem cells by at least about 2-fold. ### Claim 10 (depends on 1) The method of claim 1, wherein the method further includes delivering the adult stem cells to a tissue in a subject that is in need of repair. ### Claim 11 (depends on 10) The method of claim 10, wherein the adult stem cells are enriched and delivered to said tissue in the subject intraoperatively. ### Claim 12 (depends on 10) The method of claim 10, wherein the tissue is bone tissue. ### Claim 13 A method for detecting adult stem cells in a cell population comprising: contacting the cell population with a recognition ligand specific for an ECM component that is retained to the surfaces of adult stem cells; and detecting adult stem cells in the cell population by identifying one or more cells having the recognition ligand associated therewith or bound thereto. ### Claim 14 (depends on 13) The method of claim 13, wherein the recognition ligand is a labeled recognition ligand. ### Claim 15 (depends on 14) The method of claim 14, wherein the labeled recognition ligand is a labeled antibody. ### Claim 16 (depends on 13) The method of claim 13, wherein the adult stem cells are detected in vivo. ### Claim 17 (depends on 13) The method of claim 13, wherein the adult stem cells are detected ex vivo. ### Claim 18 (depends on 17) The method of claim 17, wherein the adult stem cells are detected using an immunoassay. ### Claim 19 (depends on 17) The method of claim 17, wherein the adult stem cells are detected using flow cytometry. ### Claim 20 (depends on 13) The method of claim 13, wherein the adult stem cells are detected in connective tissue. ### Claim 21 (depends on 13) The method of claim 13, wherein the adult stem cells are connective tissue progenitor cells. ### Claim 22 (depends on 13) The method of claim 13, wherein said ECM component is hyaluronan. ### Claim 23 (depends on 22) The method of claim 22, wherein said recognition ligand is a hyaluronan binding protein. ### Claim 24 (depends on 13) The method of claim 13, wherein said one or more cells having the recognition ligand associated therewith or bound thereto are further characterized to determine if they have other properties of adult stem cells. ### Claim 25 A method of identifying an ECM component marker associated with a particular cell type, comprising: obtaining a population of cells including one or more cells of the desired type from animal tissue; contacting the population of cells with a recognition ligand specific for a particular ECM component; enriching the cells that have the recognition ligand associated therewith or bound thereto; and determining if the enriched cells have the properties of the desired cell type. ### Claim 26 (depends on 25) The method of claim 25, wherein cells of the desired type are adult stem cells. ### Claim 27 (depends on 25) The method of claim 25, wherein the adult stem cells comprise connective tissue progenitor cells. ### Claim 28 (depends on 25) The method of claim 25, wherein said properties comprise proliferation that is different from that of non-adult stem cells. ### Claim 29 A method for enriching adult stem cells, comprising: obtaining a population of cells including one or more adult stem cells from animal tissue; contacting the population of cells with a recognition ligand specific for an EMC component or antigen that is present to a lesser degree on adult stem cells than on other cells in the population; and using a separation method to remove cells that are bound to a recognition ligand from the adult stem cells that are not bound to the recognition ligand. ### Claim 30 (depends on 29) The method of claim 29, wherein said ECM component or antigen is absent from adult stem cells in the population. ### Claim 31 (depends on 29) The method of claim 29, wherein the separation method comprises magnetic separation. ### Claim 32 (depends on 31) The method of claim 31, wherein the separation method further comprises selective retention using a porous matrix. ### Claim 33 (depends on 29) The method of claim 29, wherein the separation method comprises affinity column methods. ### Claim 34 (depends on 29) The method of claim 29, wherein the adult stem cells comprise connective tissue progenitor cells. ### Claim 35 (depends on 29) The method of claim 29, wherein the method enriches the prevalence of adult stem cells in said population by at least about 2-fold.