Stem cell material and method of manufacturing

Abstract

Aspects of the invention relate to a novel mesenchymal stem cell line (hb-MSC), a culture medium conditioned by the hb-MSC line, and various hb-MSC compositions. The hb-MSC composition may include a plurality of hb-MSCs, an hb-MSC conditioned medium, or a combination thereof. The hb-MSC composition may also include an appropriate carrier. Also described are methods of use for the hb-MSC cells, the conditioned medium and compositions thereof.

What is claimed is: 
     
         1 . A composition comprising: a conditioned medium obtained by culturing a population of non-fetal mesenchymal stem cells, wherein the conditioned medium contains a mixture of factors comprising transforming growth factor-β1 (TGF-β1), transforming growth factor-β2 (TGF-β2), and vascular endothelial growth factor (VEGF), and wherein the total concentration of the factors present in the medium is at least 12,150 pg/ml. 
     
     
         2 . The composition of  claim 1 , wherein the concentration of VEGF is at least 4000 pg/ml, the concentration of TGF-β1 is at least 750 pg/ml, and the concentration of TGF-β2 is at least 250 pg/ml. 
     
     
         3 . The composition of  claim 1 , wherein the total concentration of TGF-β1, TGF-β2 and VEGF in the conditioned medium is at least 12,150 pg/ml. 
     
     
         4 . The composition of  claim 1 , wherein the mixture of factors further comprises growth-regulated oncogene/keratinocyte chemoattractant (GRO/KC), and the concentration of GRO/KC is at least 150 pg/ml. 
     
     
         5 . The composition of  claim 1 , wherein the VEGF is at a concentration ranging from 760 to 15800 pg/ml, the TGF-β1 is at a concentration of at least 67 pg/ml, and the TGF-β2 is at a concentration of at least 170 pg/ml. 
     
     
         6 . The composition of  claim 1 , wherein the mixture of factors further comprises IL-10 and IP-10. 
     
     
         7 . The composition of  claim 1 , wherein the mesenchymal stem cells are derived from bone marrow, and wherein the conditioned medium contains less than 50 pg/ml of osteoprotegerin (OPG). 
     
     
         8 . The composition of  claim 1 , further comprising at least one of a carrier, a preservative, and a surfactant, wherein the composition is characterized in that it is free of the cells. 
     
     
         9 . The composition of  claim 8 , wherein the carrier is selected from the group consisting of a liquid, a cream, an aerosol, a lotion, an ointment, and a hydrogel. 
     
     
         10 . The composition of  claim 8 , wherein the preservative is one or more of thimerosol, cresols, formalin, benzalkonium chloride or benzyl alcohol, and wherein the surfactant is triton X-100. 
     
     
         11 . The composition of  claim 1 , wherein the non-fetal mesenchymal stem cells are characterized by positive expression of CD29 and CD44 markers and negative expression of CD11b and CD45 markers. 
     
     
         12 . The composition of  claim 1 , wherein at least 90% of the non-fetal mesenchymal stem cells exhibit a karyotype of 2 n=42. 
     
     
         13 . The composition of  claim 1 , wherein the non-fetal mesenchymal stem cells are from a stem cell line deposited with Russian National Collection of Industrial Microorganisms (VKPM) as accession number H-154. 
     
     
         14 . A composition comprising, a mixture of factors comprising VEGF, TGF-β1, and TGF-β2, wherein the factors are present in the mixture at a concentration of at least 12,150 pg/ml, and a carrier. 
     
     
         15 . The composition of  claim 14 , wherein the factors are present at concentrations of: at least 4000 pg/ml VEGF; at least 750 pg/ml TGF-β1; and at least 250 pg/ml TGF-β2. 
     
     
         16 . The composition of  claim 14 , wherein the VEGF is at a concentration ranging from 760 to 15800 pg/ml, the TGF-β1 is at a concentration of at least 67 pg/ml, and the TGF-β2 is at a concentration of at least 170 pg/ml. 
     
     
         17 . The composition of  claim 14 , wherein the mixture further comprises IL-10 and IP-10. 
     
     
         18 . The composition of  claim 14 , wherein the mixture further comprises GRO/KC at a concentration of at least 150 pg/ml. 
     
     
         19 . The composition of  claim 14 , further comprising a preservative and a surfactant. 
     
     
         20 . The composition of  claim 19 , wherein the carrier is selected from the group consisting of a liquid, a cream, an aerosol, a lotion, an ointment, and a hydrogel. 
     
     
         21 . A method for anti-aging treatment of skin, the method comprising applying topically to skin of a subject an effective amount of the composition of  claim 14 . 
     
     
         22 . A composition comprising: a conditioned medium obtained by culturing a population of non-fetal mesenchymal stem cells, wherein the conditioned medium contains at least 760 pg/ml of vascular endothelial growth factor (VEGF) and at least 150 pg/ml of growth-regulated oncogene/keratinocyte chemoattractant (GRO/KC). 
     
     
         23 . A method for providing a therapeutic application to a subject, said method comprising administering a therapeutically effective amount of the composition of  claim 22  to a subject in need thereof. 
     
     
         24 . The method of  claim 23 , wherein the therapeutic application is one or more applications selected from the group consisting of burn treatment, skin care, angiogenesis, vasculogenesis, healing of organs or tissue, tissue inflammation, bacterial infections, wound applications, diabetes, pharmaceutical and ophthalmological applications, scar reductions, stimulating hair growth, in immunotherapy applications and for immunocorrection therapy, skin, bone marrow, or organ transplants, and the treatment of organs or tissue.