ADULT MESENCHYMAL STEM CELL (MSC) COMPOSITIONS AND METHODS FOR PREPARING THE SAME
First Claim
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1. A composition comprising an isolated population of mesenchymal stem/progenitor cells (MSPCs), wherein the mesenchymal stem/progenitor cells (MSPCs) are of a cell surface antigenic profile CD34(−
- )/CD133(−
)/CD45(−
)/CD73(+)/CD90(+)/CD105(+)/CD44(−
).
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Abstract
The described invention provides a composition comprising mesenchymal progenitor cells (MPC) and processes for isolating or enriching the mesenchymal progenitor cells (MPCs) having a cell surface antigenic profile of CD34(−)/CD133(−)/CD45(−)/CD73(+)/CD90(+)/CD105(+)/CD44(−). The described invention also provides methods for differentiating the mesenchymal progenitor cells (MPCs) into various cell types.
9 Citations
49 Claims
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1. A composition comprising an isolated population of mesenchymal stem/progenitor cells (MSPCs), wherein the mesenchymal stem/progenitor cells (MSPCs) are of a cell surface antigenic profile CD34(−
- )/CD133(−
)/CD45(−
)/CD73(+)/CD90(+)/CD105(+)/CD44(−
). - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 45, 46)
- )/CD133(−
-
18. A method for isolating and purifying a population of mesenchymal stem/progenitor cells (MSPCs) of a cell surface antigenic profile CD34(−
- )/CD133(−
)/CD45(−
)/CD73(+)/CD90(+)/CD105(+)/CD44(−
), wherein the method is based on cluster of differentiation (CD) molecules on a surface of a pure initial population of cells, the method comprising;(a) acquiring a source of CD34(−
)/CD133(−
)/CD45(−
)/CD73(+)/CD90(+)/CD105(+)/CD44(−
) cells from a mammal;(b) depleting CD34-positive and CD133-positive cells from the cell source of (a) to obtain a first purified cell population of a cell surface antigenic profile CD34(−
)/CD133(−
);(c) fractionating the first purified cell population of (b) using antibodies against cell surface antigens CD45, CD73, and CD90 to obtain a second purified cell population of the cell surface antigenic profile CD34(−
)/CD133(−
)/CD45(−
)/CD73(+)/CD90(+); and(d) further fractionating the second purified cell population of (c) using antibodies against cell surface antigens CD105 and CD44 to obtain a third purified cell population of the cell surface antigenic profile CD34(−
)/CD133(−
)/CD45(−
)/CD73(+)/CD90(+)/CD105(+)/CD44(−
);wherein the method does not employ adherent culture of an unfractionated mononuclear cell population. - View Dependent Claims (19, 20, 21, 22, 23, 37, 39, 40, 41, 48, 49)
- )/CD133(−
-
24. A method for obtaining an enriched population of mesenchymal stem/progenitor cells (MSPCs) using a size-based elutriation technique, wherein the size-based elutriation technique comprises flowing the sample obtained from a subject through a series of increasing flow rates in an elutriation device, and wherein each flow rate in the series of increasing flow rates collects a different population of cells in each flow rate fraction, the method comprising:
-
(1) acquiring a sample comprising CD34(−
)/CD133(−
)/CD45(−
)/CD73(+)/CD90(+)/CD105(+)/CD44(−
) cells from a mammal(2) flowing the sample at a first flow rate, wherein the first flow rate allows a first flow rate fraction comprising cells that are smaller than the mesenchymal stem/progenitor cells (MSPCs) in the sample to flow through the elutriation device and to be collected in a first cell collection bag of the elutriation device; (3) increasing the first flow rate to a second flow rate, wherein the second flow rate allows a second flow rate fraction comprising the mesenchymal stem/progenitor cells (MSPCs) in the sample to flow through the elutriation device and to be collected in a second cell collection bag of the elutriation device, and wherein the second flow rate fraction collected in the second cell collection bag comprises the enriched population of the mesenchymal stem/progenitor cells (MSPCs); (4) recirculating the sample comprising non-retained cells through the elutriation apparatus; and (5) optionally increasing the second flow rate to a third flow rate, wherein the third flow rate allows a third flow rate fraction comprising cells that are larger than the mesenchymal stem/progenitor cells (MSPCs) in the sample to flow through the elutriation device and to be collected in a third cell collection bag of the elutriation device. - View Dependent Claims (25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 38, 47)
-
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42. A method for differentiating a mesenchymal stem/progenitor cells (MSPCs) into a neuron-like cell population, the method comprising:
-
(a) culturing a population of mesenchymal stem/progenitor cells (MSPCs) of a cell surface antigenic profile CD34(−
)/CD133(−
)/CD45(−
)/CD73(+)/CD90(+)/CD105(+)/CD44(−
) in a chemically defined growth medium devoid of an animal serum;(b) generating a spheroid by plating the mesenchymal stem/progenitor cells (MPCs) on a low-attachment plate; (c) plating and culturing the mesenchymal stem/progenitor cells (MSPCs) from the spheroid in (b) in a defined medium for neuronal differentiation; and (d) obtaining the neuron-like cell population differentiated from the mesenchymal stem/progenitor cells (MSPCs). - View Dependent Claims (43, 44)
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Specification
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Current AssigneeNeostem, Inc. (Lisata Therapeutics, Inc.)
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Original AssigneeNeostem, Inc. (Lisata Therapeutics, Inc.)
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InventorsMarasco, Wayne, Hall, Sean, Jiang, Yajuan
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Application NumberUS14/355,536Publication NumberTime in Patent OfficeDaysField of SearchUS Class Current424/93.7CPC Class CodesA61K 35/28 Bone marrow; Haematopoietic...C12N 2500/02 Atmosphere, e.g. low oxygen...C12N 2500/90 Serum-free medium, which ma...C12N 2533/30 Synthetic polymers thermore...C12N 5/0663 Bone marrow mesenchymal ste...C12N 5/0668 Mesenchymal stem cells from...
