MESENCHYMAL STROMAL CELLS AND USES RELATED THERETO
The present invention generally relates to novel preparations of mesenchymal stromal cells (MSCs) derived from hemangioblasts, methods for obtaining such MSCs, and methods of treating a pathology using such MSCs. The methods of the present invention produce substantial numbers of MSCs having a potency-retaining youthful phenotype, which are useful in the treatment of pathologies.
1 - 125 . (canceled)
126 . A method for treating a disease or disorder, comprising administering to a subject in need thereof an effective amount of mesenchymal stromal cells, wherein the mesenchymal stromal cells have replicative capacity to undergo at least 10 population doublings in cell culture with less than 25 percent of the cells undergoing cell death, senescing or differentiating into non-MSC cells by the tenth doubling.
127 . The method of claim 126 , wherein the mesenchymal stromal cells are provided as a preparation comprising at least 10 6 mesenchymal stromal cells and a pharmaceutically acceptable carrier.
128 . The method of claim 127 , wherein the preparation comprises less than one percent of a cell type other than a mesenchymal stromal cell.
129 . The method of claim 126 , wherein the mesenchymal stromal cells are HLA-genotypically identical or genomically identical.
130 . The method of claim 126 , wherein at least 30% of the mesenchymal stromal cells are positive for CD10.
131 . The method of claim 126 , wherein at least 60% of the mesenchymal stromal cells are positive for markers CD73, CD90, CD105, CD13, CD29, CD44, CD166 and HLA-ABC.
132 . The method of claim 126 , wherein less than 30% of the mesenchymal stromal cells are positive for markers CD31, CD34, CD45, CD133, FGFR2, CD271, Stro-1, CXCR4 and TLR3.
133 . The method of claim 126 , wherein the mesenchymal stromal cells have a statistically significant decreased content and/or enzymatic activity, relative to mesenchymal stromal cell preparations derived from bone marrow that have undergone five population doublings, of proteins involved in one or more of (i) cell cycle regulation and cellular aging, (ii) cellular energy and/or lipid metabolism, and (iii) apoptosis.
134 . The method of claim 126 , wherein the mesenchymal stromal cells have a statistically significant increased content and/or enzymatic activity of proteins involved in cytoskeleton structure and cellular dynamics relating thereto, relative to mesenchymal stromal cells derived from bone marrow.
135 . The method of claim 126 , wherein the mesenchymal stromal cells do not undergo more than a 75 percent increase in cells having a forward-scattered light value, measured by flow cytometry, greater than 5,000,000 over 10 population doublings in culture.
136 . The method of claim 126 , wherein the mesenchymal stromal cells, in a resting state, express mRNA encoding interleukin-6 (IL-6) at a level which is less than ten percent of the IL-6 mRNA level expressed by mesenchymal stromal cells, in a resting state, derived from bone marrow or adipose tissue.
137 . The method of claim 127 , wherein the preparation comprises at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% mesenchymal stromal cells.
138 . The method of claim 126 , wherein at least 50% of the mesenchymal stromal cells are positive for
(i) at least one of CD10, CD24, IL-11, AIRE-1, ANG-1, CXCL1, CD105, CD73 and CD90; (ii) at least one of CD10, CD24, IL-11, AIRE-1, ANG-1, CXCL1, CD105, CD73, CD90, CD105, CD13, CD29, CD44, CD166, CD274, and HLA-ABC;
(iii) CD105, CD73 and/or CD90; or
(iv) any combination thereof.
139 . The method of claim 127 , wherein the mesenchymal stromal cells are not derived from bone marrow or cord blood, and the potency of the preparation in an immune regulatory assay is greater than the potency of a preparation of bone marrow-derived or cord blood-derived mesenchymal stromal cells.
140 . The method of claim 139 , wherein potency is assayed by an immune regulatory assay that determines the EC50 dose.
141 . The method of claim 127 , wherein the preparation retains between about 50 and 100% of its proliferative capacity after ten population doublings.
142 . The method of claim 126 , wherein the mesenchymal stromal cells are mitotically inactivated.
143 . The method of claim 126 , wherein the disease or disorder is selected from multiple sclerosis, systemic sclerosis, hematological malignancies, myocardial infarction, organ transplantation rejection, chronic allograft nephropathy, cirrhosis, liver failure, heart failure, GvHD, tibial fracture, left ventricular dysfunction, leukemia, myelodysplastic syndrome, Crohn's disease, diabetes, chronic obstructive pulmonary disease, osteogenesis imperfecta, homozygous familial hypocholesterolemia, treatment following meniscectomy, adult periodontitis, vasculogenesis in patients with severe myocardial ischemia, spinal cord injury, osteodysplasia, critical limb ischemia, diabetic foot disease, primary Sjogren's syndrome, osteoarthritis, cartilage defects, multisystem atrophy, amyotropic lateral sclerosis, cardiac surgery, refractory systemic lupus erythematosis, living kidney allografts, nonmalignant red blood cell disorders, thermal burn, Parkinson's disease, microfractures, epidermolysis bullosa, severe coronary ischemia, idiopathic dilated cardiomyopathy, osteonecrosis femoral head, lupus nephritis, bone void defects, ischemic cerebral stroke, after stroke, acute radiation syndrome, pulmonary disease, arthritis, bone regeneration, inflammatory respiratory conditions, respiratory conditions due to an acute injury, Adult Respiratory Distress Syndrome, post-traumatic Adult Respiratory Distress Syndrome, transplant lung disease, Chronic Obstructive Pulmonary Disease, emphysema, chronic obstructive bronchitis, bronchitis, an allergic reaction, damage due to bacterial pneumonia, damage due to viral pneumonia, asthma, exposure to irritants, tobacco use, atopic dermatitis, allergic rhinitis, hearing loss, autoimmune hearing loss, noise-induced hearing loss, psoriasis or any combination thereof.
144 . The method of claim 126 , wherein the disease or disorder is an autoimmune disease or disorder.
145 . The method of claim 144 , wherein the autoimmune disease or disorder is selected from the group consisting of multiple sclerosis, lupus and Crohn's disease.
146 . The method of claim 126 , wherein the disease or disorder is an inflammatory disease or disorder.
147 . The method of claim 144 , wherein the inflammatory disease or disorder is uveitis.
148 . The method of claim 126 , further comprising transplantation of other cells or tissues.
149 . The method of claim 148 , further comprising transplantation of retinal, RPE, corneal, neural, immune, bone marrow, liver, or pancreatic cells.
150 . The method of claim 126 , wherein the mesenchymal stromal cells are provided in a formulation that is suitable for use in a human patient.
151 . The method of claim 126 , wherein the mesenchymal stromal cells are provided in a formulation that is pyrogen-free and/or pathogen-free.
152 . The method of claim 126 , wherein the mesenchymal stromal cells are generated in vitro from pluripotent cells.