Conditioned cell culture medium compositions and methods of use

Abstract

Novel products comprising conditioned cell culture medium compositions and methods of use are described. The conditioned cell medium compositions of the invention may be comprised of any known defined or undefined medium and may be conditioned using any eukaryotic cell type. Once the cell medium of the invention is conditioned, it may be used in any state. Physical embodiments of the conditioned medium include, but are not limited to, liquid or solid, frozen, lyophilized or dried into a powder. Additionally, the medium is formulated with a pharmaceutically acceptable carrier as a vehicle for internal administration, applied directly to a food item or product, or formulated with a salve or ointment for topical applications. Also, the medium may be further processed to concentrate or reduce one or more factors or components contained within the medium.

What is claimed is: 
     
         1 . A conditioned medium for making a composition suitable for topical application, the conditioned medium made by a process comprising the steps of:
 (a) inoculating fetal fibroblasts or embryonic stem cells onto a substrate in a nutrient medium;   (b) culturing the fetal fibroblasts or embryonic stem cells in a two dimensional culture so that the fetal fibroblasts or embryonic stem cells secrete a growth factor and a collagen into the nutrient medium, until the nutrient medium becomes a conditioned medium; and   (c) removing the conditioned medium from the cultured cells.   
     
     
         2 . The conditioned medium for making a composition suitable for topical application of  claim 1 , wherein the growth factor is keratinocyte growth factor. 
     
     
         3 . The conditioned medium for making a composition suitable for topical application of  claim 1 , wherein the substrate comprises beads. 
     
     
         4 . The conditioned medium for making a composition suitable for topical application of  claim 1 , wherein during the culturing step the fetal fibroblasts or embryonic stem cells are subjected to hypoxic stress to thereby increase the secretion of the growth factor. 
     
     
         5 . The conditioned medium for making a composition suitable for topical application of  claim 1 , further comprising the step of concentrating the conditioned medium removed from the cultured cells. 
     
     
         6 . The conditioned medium for making a composition suitable for topical application of  claim 1 , wherein the conditioned medium removed from the cultured cells is combined with an acceptable carrier. 
     
     
         7 . The conditioned medium for making a composition suitable for topical application of  claim 1 , wherein the growth factor and the collagen is at a concentration sufficient to treat a cosmetic defect when applied topically. 
     
     
         8 . A conditioned medium for making a composition suitable for topical application, the conditioned medium made by a process comprising the steps of:
 (a) inoculating mesenchymal cells onto a substrate in a nutrient medium;   (b) culturing the mesenchymal cells in a two dimensional culture so that the mesenchymal cells secrete a growth factor and a collagen into the nutrient medium, until the nutrient medium becomes a conditioned medium; and   (c) removing the conditioned medium from the cultured cells.   
     
     
         9 . The conditioned medium for making a composition suitable for topical application of  claim 8 , wherein the growth factor is keratinocyte growth factor. 
     
     
         10 . The conditioned medium for making a composition suitable for topical application of  claim 8 , wherein the substrate comprises beads. 
     
     
         11 . The conditioned medium for making a composition suitable for topical application of  claim 8 , wherein during the culturing step the mesenchymal cells are subjected to hypoxic stress to thereby increase the secretion of the growth factor. 
     
     
         12 . The conditioned medium for making a composition suitable for topical application of  claim 8 , further comprising concentrating the conditioned medium removed from the cultured cells. 
     
     
         13 . The conditioned medium for making a composition suitable for topical application of  claim 8 , wherein the growth factor and the collagen is at a concentration sufficient to treat a cosmetic defect when applied topically. 
     
     
         14 . A conditioned medium for making a composition suitable for topical application, the conditioned medium made by a process comprising the steps of:
 (a) inoculating fetal fibroblasts or embryonic stem cells onto a substrate in a nutrient medium;   (b) culturing the fetal fibroblasts or embryonic stem cells in a two dimensional culture so that the fetal fibroblasts or embryonic stem cells secrete a growth factor and a collagen into the nutrient medium, until the nutrient medium becomes a conditioned medium;   (c) subjecting the fetal fibroblasts or embryonic stem cells during the culturing step to hypoxic stress to increase the secretion of the growth factor; and   (d) removing the conditioned medium from the cultured cells.   
     
     
         15 . The conditioned medium for making a composition suitable for topical application of  claim 14 , wherein the substrate comprises beads. 
     
     
         16 . The conditioned medium for making a composition suitable for topical application of  claim 14 , further comprising the step of concentrating the conditioned medium removed from the cultured cells.