Adult stem cells derived from human skin dermis

Abstract

Provided in the present invention are adult stem cells derived from human skin dermis, and a method for isolating same. Further provided in the present invention are osteoblastic cells and adipocytes differentiated from the adult stem cells derived from human skin dermis, and a differentiation method therefor. Further provided in the present invention is a composition for osteogenesis or lipogenesis containing the stem cells, osteoblastic cells, or adipocytes. The isolation method of the present invention enables the adult stem cells derived from human skin dermis to be obtained in an easy and simple manner at a high yield rate. Genes and growth factors which are specifically expressed in the adult stem cells derived from human skin dermis isolated using the method can be separated, identified, and used later.

1 . Human skin dermis-derived adult stem cells wherein one or more gene selected from a group consisting of Sox2 (SRY (sex determining region Y)-box 2) and S100b (S100 calcium binding protein B) is overexpressed as compared to human skin dermis-derived fibroblasts. 
     
     
         2 . The human skin dermis-derived adult stem cells according to  claim 1 , wherein the stem cells are obtained by subculturing human skin dermis-derived fibroblasts, reacting the cells with gelatin or type 4 collagen and separating the cells adhering to the gelatin or type 4 collagen. 
     
     
         3 . The human skin dermis-derived adult stem cells according to  claim 2 , wherein the stem cells are obtained by reacting the fibroblasts with gelatin or type 4 collagen for 1-5 minutes. 
     
     
         4 . The human skin dermis-derived adult stem cells according to  claim 2 , wherein the gelatin is one dissolved in distilled water to a concentration of 0.1-1 wt % and the type 4 collagen is one dissolved in distilled water to a concentration of 10-30 μg/mL. 
     
     
         5 . The human skin dermis-derived adult stem cells according to  claim 2 , wherein the gelatin or the type 4 collagen is one coated on a substrate at 0-10° C. for 16-24 hours. 
     
     
         6 . The human skin dermis-derived adult stem cells according to  claim 1 , wherein the stem cells are ones wherein one or more growth factor selected from a group consisting of EGF (epidermal growth factor), FGF4 (fibroblast growth factor 4), PDGF-AA (platelet-derived growth factor-AA), VEGFR-2 (vascular endothelial growth factor receptor 2), VEGFR-3 (vascular endothelial growth factor receptor 3) and VEGF-D (vascular endothelial growth factor D) is overexpressed as compared to human skin dermis-derived fibroblasts. 
     
     
         7 . The human skin dermis-derived adult stem cells according to  claim 1 , wherein the stem cells are stem cells of Accession No: KCTC11995BP. 
     
     
         8 . Differentiated cells differentiated from the human skin dermis-derived adult stem cells according to  claim 1 ,
 Wherein the differentiated cells are osteoblasts or adipocytes.   
     
     
         9 . The differentiated cells according to  claim 8 , wherein the osteoblasts are ones wherein one or more gene selected from a group consisting of OGN (osteoglycin) and ACAN (aggrecan) is overexpressed as compared to skin dermis-derived fibroblasts. 
     
     
         10 . The differentiated cells according to  claim 8 , wherein the adipocytes are ones wherein one or more gene selected from a group consisting of PPARG (peroxisome proliferator-activated receptor gamma), LEP (leptin), AdipoQ (adiponectin, C1Q and collagen domain containing) and FABP4 (fatty acid binding protein 4, adipocyte) is overexpressed as compared to skin dermis-derived fibroblasts. 
     
     
         11 . A method for ostcogenesis in a human in need thereof comprising administering a therapeutically effective amount of the human skin dermis-derived adult stem cells according to  claim 1  or osteoblasts differentiated from the human skin dermis-derived adult stem cells according to  claim 1  for osteogenesis in the human. 
     
     
         12 . The method according to  claim 11 , wherein the method is for preventing and treating osteoporosis, preventing and treating skin aging, treating skin wound, improving skin blood circulation, enhancing skin volume or skin grafting. 
     
     
         13 . A method for isolating human skin dermis-derived adult stem cells, comprising subculturing human skin dermis-derived fibroblasts, reacting the cells with gelatin or type 4 collagen and separating the cells adhering to the gelatin or type 4 collagen. 
     
     
         14 . The method for isolating human skin dermis-derived adult stem cells according to  claim 13 , wherein the fibroblasts are reacted with gelatin or type 4 collagen for 1-5 minutes. 
     
     
         15 . The method for isolating human skin dermis-derived adult stem cells according to  claim 13 , wherein the gelatin is dissolved in distilled water to a concentration of 0.1-1 wt % and the type 4 collagen is dissolved in distilled water to a concentration of 10-30 μg/mL and then reacted with the fibroblasts. 
     
     
         16 . The method for isolating human skin dermis-derived adult stem cells according to  claim 13 , wherein the gelatin or the type 4 collagen is coated on a substrate at 0-10° C. for 16-24 hours and then reacted with the fibroblasts. 
     
     
         17 . The method for isolating human skin dermis-derived adult stem cells according to  claim 13 , which further comprises confirming whether one or more gene selected from a group consisting of Sox2 (SRY (sex determining region Y)-box 2) and S100b (S100 calcium binding protein B) is overexpressed as compared to skin dermis-derived fibroblasts. 
     
     
         18 . The method for isolating human skin dermis-derived adult stem cells according to  claim 13 , which further comprises confirming whether one or more growth factor selected from a group consisting of EGF (epidermal growth factor), FGF4 (fibroblast growth factor 4), PDGF-AA (platelet-derived growth factor-AA), VEGFR-2 (vascular endothelial growth factor receptor 2), VEGFR-3 (vascular endothelial growth factor receptor 3) and VEGF-D (vascular endothelial growth factor D) is overexpressed as compared to skin dermis-derived fibroblasts. 
     
     
         19 . A method for differentiating human skin dermis-derived adult stem cells into osteoblasts or adipocytes, the method comprising differentiating the human skin dermis-derived adult stem cells isolated by the method according to  claim 13 . 
     
     
         20 . The method for differentiating human skin dermis-derived adult stem cells according to  claim 19 , which comprises:
 differentiating the isolated stem cells in an osteogenic differentiation medium; and   confirming whether the differentiated cells overexpress one or more gene selected from a group consisting of OGN (osteoglycin) and ACAN (aggrecan) as compared to skin dermis-derived fibroblasts.   
     
     
         21 . The method for differentiating human skin dermis-derived adult stem cells according to  claim 19 , which comprises:
 differentiating the isolated stem cells in an adipogenic differentiation medium; and   confirming whether the differentiated cells overexpress one or more gene selected from a group consisting of PPARG (peroxisome pro liferator-activated receptor gamma), LEP (leptin), AdipoQ (adiponectin, C1Q and collagen domain containing) and FABP4 (fatty acid binding protein 4, adipocyte) as compared to skin dermis-derived fibroblasts.   
     
     
         22 . A method for adipogenesis in a human in need thereof comprising administering a therapeutically effective amount of the human skin dermis-derived adult stem cells according to  claim 1  or adipocytes differentiated from the human skin dermis-derived adult stem cells according to  claim 1  for adipogenesis in the human. 
     
     
         23 . The method according to  claim 22 , wherein the method is for preventing and treating osteoporosis, preventing and treating skin aging, treating skin wound, improving skin blood circulation, enhancing skin volume or skin grafting.