# US 2014/0193375 A1 · Mesenchymal Stromal Cell Populations and Methods of Making Same > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 2014/0193375 A1 - Document ID: US20140193375A1 - Status: Published application - Kind: A1 - Application number: 14/117,774 - Filed: 2012-05-18 - Published: 2014-07-10 - Earliest claimed: 2011-05-18 - Assignees: Aastrom Biosciences, Inc. - Inventors: Frank Zeigler; Ronnda L. Bartel - Web page: https://patentgrep.com/patent/US20140193375A1 ## CPC - A61K 35/28: Bone marrow; Haematopoietic stem cells; Mesenchymal stem cells of any origin, e.g. adipose-derived stem cells - A61P 1/02: Stomatological preparations, e.g. drugs for caries, aphtae, periodontitis - A61P 1/16: for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics - A61P 9/00: Drugs for disorders of the cardiovascular system - A61P 11/00: Drugs for disorders of the respiratory system - A61P 17/00: Drugs for dermatological disorders - A61P 19/00: Drugs for skeletal disorders - A61P 25/00: Drugs for disorders of the nervous system - C12N 5/0645: {Macrophages, e.g. Kuepfer cells in the liver; Monocytes} - C12N 5/0665: {Blood-borne mesenchymal stem cells, e.g. from umbilical cord blood} - C12N 5/0669: {Bone marrow stromal cells; Whole bone marrow (isolated stem cells from bone marrow C12N5/0647, C12N5/0663)} - A61K 2035/122: {for inducing tolerance or supression of immune responses} - A61K 2035/124: {the cells being hematopoietic, bone marrow derived or blood cells} - C12N 2501/51: B7 molecules, e.g. CD80, CD86, CD28 (ligand), CD152 (ligand) - C12N 2502/137: Blood-borne mesenchymal stem cells, e.g. Msc from umbilical cord blood - C12N 2509/00: Methods for the dissociation of cells, e.g. specific use of enzymes - A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D) - A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) ## Abstract The present invention provides compositions of mesenchymal stromal cells which express B7-H3, their subsequent use in tissue repair, improved methods of producing tissue repair cells and method of producing a substantially pure population of CD14+ autofluorescent macrophages. ## Claims ### Claim 1 An isolated mesenchymal stromal cell composition wherein the mesenchymal stromal cells express B7 homolog 3 (B7-H3). ### Claim 2 An isolated cell composition comprising mesenchymal stromal cells that express B7 homolog 3 (B7-H3). ### Claim 3 (depends on 2) The composition of claim 2, wherein the mesenchymal stromal cells are non-proliferative after less than 5 passages in culture. ### Claim 4 (depends on 2) The composition claim 2, wherein the mesenchymal stromal cells have been immortalized. ### Claim 5 (depends on 2) The composition of claim 2, wherein at least 90% of the cells express CD90. ### Claim 6 (depends on 2) The composition of claim 2, wherein at least 80% of the cells express CD90. ### Claim 7 (depends on 2) The composition of claim 2, wherein at least 70% of the cells express CD90. ### Claim 8 (depends on 2) The cell composition of claim 2, wherein the composition comprises less than 25% viable CD45+ cells. ### Claim 9 (depends on 2) The composition of claim 2, wherein the mesenchymal stromal cells are derived from hematopoietic cells. ### Claim 10 (depends on 2) The composition of claim 2, wherein the mesenchymal stromal cells are adherent in culture. ### Claim 11 (depends on 2) The composition of claim 2, wherein the mesenchymal stromal cells do not differentiate in culture. ### Claim 12 (depends on 2) The composition of claim 2, wherein said composition inhibits T-cell activation. ### Claim 13 (depends on 2) The composition of claim 2, wherein said composition promotes the expansion of Th2/CD8 lymphocytes. ### Claim 14 (depends on 2) The composition of claim 2, wherein the mesenchymal stromal cells are CD90+ and CD105+. ### Claim 15 (depends on 14) The composition of claim 14, wherein the mesenchymal stromal cells are CD146+ and CD73+. ### Claim 16 (depends on 2) The composition of claim 2, wherein the mesenchymal stromal cells are derived from mononuclear cells. ### Claim 17 (depends on 16) The composition of claim 16, wherein the mononuclear cells are derived from mobilized peripheral blood, bone marrow, umbilical cord blood or fetal liver. ### Claim 18 A mesenchymal stromal cell that has been genetically engineered to stably express B7 homolog 3 (B7-H3) on the surface of the cell. ### Claim 19 (depends on 2) The composition of claim 2, wherein the composition contains a) less than 2 μg/ml of bovine serum albumin; b) less than 1 μg/ml of a enzymatically active harvest reagent; and c) substantially free of mycoplasm, endotoxin, and microbial contamination. ### Claim 20 (depends on 2) A method of tissue regeneration or repair comprising administering to a patient in need thereof the composition of claim 2. ### Claim 21 (depends on 20) The method of claim 20, wherein said tissue is selected from the group consisting of cardiac tissue, bone tissue, neuronal tissue, skin tissue, lung tissue, salivary gland tissue, liver tissue, and pancreatic tissue. ### Claim 22 A method of producing a cell composition comprising a mixed population of cells of hematopoietic, mesenchymal and endothelial lineage, wherein the cell composition is characterized as containing 5-75% viable CD90+ cells with the remaining cells in said composition being CD45+, CD31+, CD14+, and auto+, comprising culturing mononuclear cells in the presence of i) B7-H3 polypeptide, ii) a V-set and Ig domain-containing 4 (VSIG4) polypeptide, or iii) both i) and ii). ### Claim 23 A method of producing a substantially pure population of CD14+ autofluorescent macrophages comprising culturing mononuclear cells in the presence of i) a B7-H3 polypeptide, ii) a V-set and Ig domain-containing 4 (VSIG4) polypeptide, or iii) both i) and ii); and isolating said CD14+ autofluorescent macrophages from said culture. ### Claim 24 (depends on 2) A method of producing a substantially pure population of CD14+ autofluorescent macrophages comprising culturing mononuclear cells in the presence of the composition of claim 2 and isolating said CD14+ autofluorescent macrophages from said culture. ### Claim 25 (depends on 23) The method of claim 23, wherein the macrophages express at least one of the following markers: CD45, CD163 or CD206. ### Claim 26 (depends on 22) The method of claim 22, wherein the culturing is performed by: providing a biochamber for culturing the mononuclear cells; providing a culture media for culturing the mononuclear cells within the biochamber; inoculating the biochamber with the mononuclear cells; and culturing the mononuclear cells. ### Claim 27 (depends on 26) The method of claim 26, further comprising: upon a predetermined time period of culture, displacing the culture media from the biochamber with a biocompatible first rinse solution; replacing the first rinse solution with a cell harvest enzyme solution; incubating the contents of the biochamber for a predetermined period of time, wherein during incubation, the enzyme at least dissociates the cells i) from each other, ii) from the biochamber surface, or iii) from each other and from the biochamber surface; displacing the enzyme solution with a second rinse solution, wherein upon the enzyme being displaced, the chamber is substantially filled with the second rinse solution; displacing a portion of the second rinse solution with a gas to obtain a predetermined reduced liquid volume in the chamber; agitating the chamber to bring settled cells into suspension; and draining the solution with suspended cells into a cell collection container. ### Claim 28 (depends on 23) The method of claim 23, wherein the culturing is performed by: providing a biochamber for culturing the mononuclear cells; providing a culture media for culturing the mononuclear cells within the biochamber; inoculating the biochamber with the mononuclear cells; and culturing the mononuclear cells. ### Claim 29 (depends on 28) The method of claim 28, further comprising: upon a predetermined time period of culture, displacing the culture media from the biochamber with a biocompatible first rinse solution; replacing the first rinse solution with a cell harvest enzyme solution; incubating the contents of the biochamber for a predetermined period of time, wherein during incubation, the enzyme at least dissociates the cells i) from each other, ii) from the biochamber surface, or iii) from each other and from the biochamber surface; displacing the enzyme solution with a second rinse solution, wherein upon the enzyme being displaced, the chamber is substantially filled with the second rinse solution; displacing a portion of the second rinse solution with a gas to obtain a predetermined reduced liquid volume in the chamber; agitating the chamber to bring settled cells into suspension; and draining the solution with suspended cells into a cell collection container. ### Claim 30 (depends on 24) The method of claim 24, wherein the culturing is performed by: providing a biochamber for culturing the mononuclear cells; providing a culture media for culturing the mononuclear cells within the biochamber; inoculating the biochamber with the mononuclear cells; and culturing the mononuclear cells. ### Claim 31 (depends on 30) The method of claim 30, further comprising: upon a predetermined time period of culture, displacing the culture media from the biochamber with a biocompatible first rinse solution; replacing the first rinse solution with a cell harvest enzyme solution; incubating the contents of the biochamber for a predetermined period of time, wherein during incubation, the enzyme at least dissociates the cells i) from each other, ii) from the biochamber surface, or iii) from each other and from the biochamber surface; displacing the enzyme solution with a second rinse solution, wherein upon the enzyme being displaced, the chamber is substantially filled with the second rinse solution; displacing a portion of the second rinse solution with a gas to obtain a predetermined reduced liquid volume in the chamber; agitating the chamber to bring settled cells into suspension; and draining the solution with suspended cells into a cell collection container. ### Claim 32 (depends on 24) The method of claim 24, wherein the macrophages express at least one of the following markers: CD45, CD163 or CD206.