CLINICAL DERIVATIONS OF AN ALLOGENIC CELL AND THERAPEUTIC USES
Various cells, stem cells, and stem cell components, including associated methods of generating and using such cells are provided. In one aspect, for example, an isolated cell that is capable of self-renewal and culture expansion and is obtained from a subepithelial layer of a mammalian umbilical cord tissue. Such an isolated cell expresses at least three cell markers selected from CD29, CD73, CD90, CD166, SSEA4, CD9, CD44, CD146, or CD105, and does not express at least three cell markers selected from CD45, CD34, CD14, CD79, CD106, CD86, CD80, CD19, CD117, Stro-1, or HLA-DR.
1 . A method of preparing an isolated cell, comprising:
placing a subepithelial layer of a mammalian umbilical cord tissue in direct contact with a growth substrate; and
culturing the subepithelial layer such that the isolated cell from the subepithelial layer is capable of self-renewal and culture expansion,
wherein the isolated cell expresses at least three cell markers selected from the group consisting of CD29, CD73, CD90, CD166, SSEA4, CD9, CD44, CD146, or CD105, and
wherein the isolated cell does not express NANOG and at least five cell markers selected from the group consisting of CD45, CD34, CD14, CD79, CD106, CD86, CD80, CD19, CD117, Stro-1, or HLA-DR.
2 . The method of claim 1 , wherein the isolated cell expresses CD29, CD73, CD90, CD166, SSEA4, CD9, CD44, CD146, and CD105.
3 . The method of claim 1 , wherein the isolated cell does not express CD45, CD34, CD14, CD79, CD106, CD86, CD80, CD19, CD117, Stro-1, and HLA-DR.
4 . The method of claim 1 , wherein the isolated cell is positive for SOX2.
5 . The method of claim 1 , wherein the isolated cell is positive for OCT4.
6 . The method of claim 1 , wherein the isolated cell is positive for SOX2 and OCT4.
7 . The method of claim 1 , wherein the wherein the isolated cell is capable of differentiation into a cell type selected from the group consisting of adipocytes, chondrocytes, osteocytes, cardiomyocytes, endothelial cells, and myocytes.
8 . The method of claim 1 , wherein the isolated cell produces exosomes expressing CD63, CD9, or CD63 and CD9.
9 . The method of claim 1 , wherein culturing comprises culturing in a culture media that is free of animal components.
10 . The method of claim 1 , comprising culturing differentiated cells derived from the isolated cell to form a culture of differentiated cells including a cell type selected from the group consisting of adipocytes, chondrocytes, osteocytes, cardiomyocytes, endothelial cells, myocytes, and combinations thereof.
11 . The method of claim 1 that has been differentiated into an adipocyte cell.
12 . The method of claim 1 that has been differentiated into a chondrocyte cell.
13 . The method of claim 1 that has been differentiated into an osteocyte cell.
14 . The method of claim 1 that has been differentiated into a cardiomyocyte cell.
15 . The method of claim 1 that has been expanded into a cell culture.
16 . The method of claim 1 , wherein:
placing the subepithelial layer includes:
dissecting the subepithelial layer from the umbilical cord, wherein dissecting the subepithelial layer further includes removing Wharton's Jelly from the umbilical cord, and
placing the dissected subepithelial layer interior side down on a substrate such that an interior side of the subepithelial layer is in contact with the substrate; and
culturing the subepithelial layer includes removing explants for primary cell expansion.
17 . The method of claim 16 , wherein the subepithelial layer is cultured in a culture medium comprising a platelet lysate.
18 . The method of claim 16 , wherein the subepithelial layer is cultured in a normoxic environment.
19 . The method of claim 16 , wherein the subepithelial layer is cultured in a hypoxic environment.
20 . The method of claim 16 , wherein culturing the subepithelial layer, subculturing the explants removed for primary cell expansion, or both is performed without the use of any enzymes.
21 . The culture of differentiated cells prepared in accordance with the method of claim 10 .
22 . A method of treating a medical condition responsive to treatment with the culture of differentiated cells of claim 21 , comprising introducing stem cells from the culture of differentiated cells into an individual having the medical condition.
23 . The method of claim 22 , wherein introducing the stem cells into the individual includes administering at least a portion of the stem cells from the culture of differentiated cells to the individual.
24 . The method of claim 23 , wherein the medical condition includes one or more of COPD, diabetes, ischemia, osteoarthritis, orthopedic damage, liver damage, chronic refractory angina, erectile dysfunction, spinal cord injuries, herniated disks, congestive heart failure, asthma, emphysema, wounds, acute radiation syndrome, autoimmune disorders, ischemic organ beds, or graft vs. host disease.
25 . The method of claim 23 , wherein the portion of the stem cells include stem cells differentiated into adipocyte cells.
26 . The method of claim 25 , wherein the medical condition includes one or more of COPD, diabetes, ischemia, osteoarthritis, orthopedic damage, liver damage, chronic refractory angina, erectile dysfunction, spinal cord injuries, herniated disks, congestive heart failure, asthma, emphysema, wounds, acute radiation syndrome, autoimmune disorders, ischemic organ beds, or graft vs. host disease.
27 . The method of claim 25 , wherein the medical condition is COPD.
28 . The method of claim 25 , wherein administering the at least the portion of the stem cells includes administering the adipocyte cells to the individual.
29 . The method of claim 28 , wherein administering the adipocyte cells to the individual includes:
intravenously administering the adipocyte cells to the individual to deliver the adipocyte cells to a lower half of the subject's lung;
administering the adipocyte cells in an aerosol to the individual via ventilation to an upper half of the subject's lung; or
both.
30 . The method of claim 29 , wherein the aerosol has an average droplet or particle size from about 6 to about 200 microns.
31 . The method of claim 27 , wherein administering includes coadministering exosomes, cell lysates, protein extracts derived from cell culture, or a combination thereof.
32 . The method of claim 23 , wherein administering the stem cells from the culture of differentiated cells into the individual further includes retrograde or antegrade delivery of the cells into an organ of the individual.
33 . The method of claim 32 , wherein the organ includes one or more of a heart, a liver, a kidney, the brain, or a pancreas.