Compositions comprising conditioned cell culture media and uses thereof

Abstract

The invention relates to compositions comprising cell culture medium conditioned by cells grown in three-dimensional culture. The cells used to condition the medium may be genetically modified to alter the concentration of growth factors and antioxidants in the medium. The conditioned cell medium (conditioned medium) may be used for at least one of cosmetic applications, cosmeceutical applications, and pharmaceutical applications, among other things. The invention also relates to proteins comprising a heterologous sequence that enhances cell penetration. The invention also relates to cells comprising DNA encoding such proteins. Methods for preparing the inventive compounds are also provided.

1 - 62 . (canceled)  
     
     
         63 . A growth factor comprising a heterologous peptide sequence that enhances cell penetration.  
     
     
         64 . The growth factor of  claim 63 , wherein the growth factor comprises at least one of insulin, insulin-like growth factor (IGF), nerve growth factor, VEGF, keratinocyte growth factor (KGF), fibroblast growth factor (FGF), platelet-derived growth factor (PDGF), hepatocyte growth factor (HGF), transforming growth factor alpha (TGFα), transforming growth factor beta (TGFβ), epidermal growth factor (EGF), granulocyte-macrophage colony-stimulating factor (GM-CSF), granulocyte colony-stimulating factor (G-CSF), interleukin-6 (IL-6), and interleukin-8 (IL-8); and wherein the heterologous peptide sequence comprises at least one of the following sequences: SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, SEQ ID NO:17, SEQ ID NO:18, and SEQ ID NO:19.  
     
     
         65 . An antioxidant comprising a heterologous peptide sequence that enhances cell penetration.  
     
     
         66 . The antioxidant of  claim 65 , wherein the antioxidant comprises at least one of cysteine, cystine, ascorbic acid, glutathione, glutathione disulfide, glutathione peroxidase, glutathione reductase, glutathione disulfide, superoxide dismutase, catalase, vitamin E, ascorbic acid, ubiquinol 9, and ubiquinone 9; and wherein the heterologous peptide sequence comprises at least one of the following sequences: SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, SEQ ID NO:17, SEQ ID NO:18, and SEQ ID NO:19.  
     
     
         67 . An extracellular matrix component comprising a heterologous peptide sequence that enhances cell penetration.  
     
     
         68 . The extracellular matrix component of  claim 67 , wherein the extracellular matrix component comprises at least one of: at least one glycoprotein, at least one proteoglycan, and at least one glycosaminoglycan; and wherein the heterologous peptide sequence comprises at least one of the following sequences: SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, SEQ ID NO:17, SEQ ID NO:18, and SEQ ID NO:19.  
     
     
         69 . A cell comprising DNA encoding a growth factor comprising a heterologous peptide sequence that enhances cell penetration.  
     
     
         70 . A cell according to  claim 69 , wherein the growth factor comprises at least one of insulin, insulin-like growth factor (IGF), nerve growth factor, VEGF, keratinocyte growth factor (KGF), fibroblast growth factor (FGF), platelet-derived growth factor (PDGF), hepatocyte growth factor (HGF), transforming growth factor alpha (TGFa), transforming growth factor beta (TGF, epidermal growth factor (EGF), granulocyte-macrophage colony-stimulating factor (GM-CSF), granulocyte colony-stimulating factor (G-CSF), interleukin-6 (IL-6), and interleukin-8 (IL-8); and wherein the heterologous peptide sequence comprises at least one of the following sequences: SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, SEQ ID NO:17, SEQ ID NO:18, and SEQ ID NO:19.  
     
     
         71 . A cell comprising DNA encoding an antioxidant comprising a heterologous peptide sequence that enhances cell penetration.  
     
     
         72 . A cell according to  claim 71 , wherein the antioxidant comprises at least one of cysteine, cystine, ascorbic acid, glutathione, glutathione disulfide, glutathione peroxidase, glutathione reductase, glutathione disulfide, superoxide dismutase, catalase, vitamin E, ascorbic acid, ubiquinol 9, and ubiquinone 9; and wherein the heterologous peptide sequence comprises at least one of the following sequences: SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, SEQ ID NO:17, SEQ ID NO:18, and SEQ ID NO:19.  
     
     
         73 . A cell comprising DNA encoding an extracellular matrix component comprising a heterologous peptide sequence that enhances cell penetration.  
     
     
         74 . A cell according to  claim 73 , wherein the extracellular matrix component comprises at least one of: at least one glycoprotein, at least one proteoglycan, and at least one glycosaminoglycan; and wherein the heterologous peptide sequence comprises at least one of the following sequences: SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:1O, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, SEQ ID NO:17, SEQ ID NO:18, and SEQ ID NO:19.