Factor rich product from umbilical cord mesenchymal stem cells

Abstract

Factor rich compositions produced from umbilical cord mesenchymal stem cells and processes for making and using same are described. The manufacturing process includes utilizing secretory UC MSCs, providing serum and growth factor free growth conditions, and performing filtration of the collected conditioned medium to obtain a clinical grade product.

What is claimed is: 
     
         1 . A factor rich composition derived from umbilical cord (UC) mesenchymal stem cells (MSCs) cultured without serum or exogenous growth factors comprising:
 a. a population of exosomes at a concentration of at least 22 billion per milliliter (ml);   b. said population of exosomes at an average size of about 133.4 nanometers (nm);   c. a growth factor selected from the group consisting of stem cell factor (SCF), vascular endothelial growth factor (VEGF), nerve growth factor (NGF) and combinations thereof;   d. said growth factors at a concentration of at least 20-2000 picogram (pg)/ml;   e. a cytokine or chemokine selected from the group consisting of interferon gamma, (INF-γ), interleukins (IL) 3, 6, 18, IL-4 7, 8, macrophage inflammatory protein-1 beta (MIP1β), monocyte chemoattractant protein-1 (MCP1) and tumor necrosis factor alpha (TNFα) and combinations thereof;   f. a glycosaminoglycan (GAG) selected from the group consisting of hyaluronic acid (HA) and chondroitin sulfate (CS);   g. a collagen selected from the group consisting of collagen I and collage III.   
     
     
         2 . The composition of  claim 1 , wherein said HA is present at a concentration of at least 1 microgram (μg)/ml. 
     
     
         3 . The composition of  claim 1 , wherein said HA is of molecular weight up to about 2500 kilodaltons (kDa). 
     
     
         4 . The composition of  claim 1 , wherein said collagen is present at a concentration of at least 0.281 microgram (mg)/ml. 
     
     
         5 . The composition of  claim 1 , wherein the product is provided in a physical state selected from the group consisting of liquid, frozen, lyophilized. 
     
     
         6 . The composition of  claim 1 , wherein the product is sterile. 
     
     
         7 . A factor rich composition made by a process including the steps of:
 isolating an umbilical cord lining mesenchymal stem cells (ULSCs) line;   culturing said ULSCs in a serum and growth factor free cell culture medium;   filtering a collected ULSCs conditioned medium three times, serially, over 0.45 μm, 0.22 μm and 0.22 μm filters.   
     
     
         8 . A process of making a factor rich composition from UC MSCs which comprises:
 a. Isolating a UC MSCs cell line;   b. Expanding a population size of said UC MSCs in a culture medium containing serum;   c. Replacing said culture medium with a culture medium free of serum and growth factors;   d. Collecting a conditioned culture medium at regular intervals;   e. Pooling said conditioned medium;   f. Filtering said conditioned medium.   
     
     
         9 . The process of  claim 8 , wherein said UC MSCs cell line is a ULSCs cell line. 
     
     
         10 . The process of  claim 8 , wherein said conditioned culture medium is collected every third day. 
     
     
         11 . The process of  claim 8 , wherein said conditioned culture medium is filtered by a technique selected from the group consisting of vacuum filtration, gravity filtration, syringe filtration, and tangential flow filtration. 
     
     
         12 . The process of  claim 8 , wherein the conditioned medium is filtered through a filter membrane selected from the group consisting of polyethersulfone (PES), polyvinylidene fluoride (PVDF), mixed cellulose esters, cellulose acetate, cellulose nitrate, or combinations thereof. 
     
     
         13 . The process of  claim 8 , wherein the conditioned medium is filtered through a 0.45 μm filter. 
     
     
         14 . The process of  claim 8 , wherein the conditioned medium is filtered through a 0.22 μm filter. 
     
     
         15 . The process of  claim 8 , wherein the conditioned medium is filtered through a 0.45 μm filter and then filtered through a 0.22 μm filter. 
     
     
         16 . The process of  claim 8 , wherein the conditioned medium is filtered through a 0.45 μm filter and then filtered through a 0.22 μm filter serially in a single step. 
     
     
         17 . The process of  claim 8 , wherein the conditioned medium is filtered through a 0.45 μm filter and then filtered through a 0.22 μm filter, and then filtered through a 0.22 μm filter serially in a single step.