Differentiation-inducing culture medium additive and use thereof

Abstract

Provided is a differentiation-inducing culture medium additive for inducing bone differentiation of at least one type of cell selected from the group consisting of a stem cell, a dental pulp cell, a periodontal ligament cell, a placenta, an amnion, and a fibroblast under a serum-free condition, and a use of the differentiation-inducing culture medium additive. The differentiation-inducing culture medium additive of the present invention for inducing differentiation of a stem cell under a serum-free condition at least contains at least one growth factor selected from the group consisting of EGF, FGF, and PDGF; dexamethasone; and β-glycerophosphate. The differentiation-inducing culture medium additive of the present invention does not require ascorbic acid 2-phosphate and ITS, which are normally essential for bone differentiation. Further, bone differentiation can be promoted by adding phospholipid.

1 ) A differentiation-inducing culture medium additive for inducing bone differentiation of at least one type of cell selected from the group consisting of a stem cell, a dental pulp cell, a periodontal ligament cell, a placenta, an amnion, and a fibroblast under a serum-free condition,
 the differentiation-inducing culture medium additive at least comprising:   at least one growth factor selected from the group consisting of EGF, FGF, and PDGF;   dexamethasone; and   β-glycerophosphate.   
     
     
         2 ) The differentiation-inducing culture medium additive according to  claim 1 , further comprising at least one phospholipid. 
     
     
         3 ) The differentiation-inducing culture medium additive according to  claim 2 , wherein the at least one phospholipid is selected from the group consisting of phosphatidic acid, lysophosphatidic acid, phosphatidylinositol, phosphatidylserine, phosphatidylethanolamine, phosphatidyl choline, and phosphatidylglycerol. 
     
     
         4 ) The differentiation-inducing culture medium additive according to  claim 2 , further comprising an oxidation inhibitor. 
     
     
         5 ) The differentiation-inducing culture medium additive according to  claim 4 , wherein the oxidation inhibitor is DL-α-tocopherol acetate (vitamin E). 
     
     
         6 ) The differentiation-inducing culture medium additive according to  claim 1 , wherein the stem cell is a mesenchymal stem cell. 
     
     
         7 ) A culture medium for inducing bone differentiation of at least one type of cell selected from the group consisting of a stem cell, a dental pulp cell, a periodontal ligament cell, a placenta, an amnion, and a fibroblast under a serum-free condition,
 the culture medium comprising:   components constituting a differentiation-inducing culture medium additive as set forth in  claim 1 ; and   a basal medium,   the culture medium containing no serum.   
     
     
         8 ) The culture medium according to  claim 7 , wherein the stem cell is a mesenchymal stem cell. 
     
     
         9 ) A culture method for inducing bone differentiation of at least one type of cell selected from the group consisting of a stem cell, a dental pulp cell, a periodontal ligament cell, a placenta, an amnion, and a fibroblast under a serum-free condition,
 the culture method comprising the step of:   differentiating at least one type of cell selected from the group consisting of the stem cell, the dental pulp cell, the periodontal ligament cell, the placenta, the amnion, and the fibroblast in the culture medium as set forth in  claim 7 .   
     
     
         10 ) The culture method according to  claim 9 , wherein the stem cell is a mesenchymal stem cell. 
     
     
         11 ) A kit for inducing bone differentiation of at least one type of cell selected from the group consisting of a stem cell, a dental pulp cell, a periodontal ligament cell, a placenta, an amnion, and a fibroblast under a serum-free condition,
 the kit comprising:   components constituting a differentiation-inducing culture medium additive as set forth in  claim 1 .   
     
     
         12 ) The kit according to  claim 11 , wherein the stem cell is a mesenchymal stem cell. 
     
     
         13 ) The culture method according to  claim 9 , further comprising the step of:
 proliferating at least one type of cell selected from the group consisting of the stem cell, the dental pulp cell, the periodontal ligament cell, the placenta, the amnion, and the fibroblast in a serum-free culture medium containing at least three growth factors selected from the group consisting of FGF, PDGF, EGF, TGF-β, and HGF and at least one phospholipid before the step of differentiating at least one type of cell selected from the group consisting of the stem cell, the dental pulp cell, the periodontal ligament cell, the placenta, the amnion, and the fibroblast in the culture medium as set forth in  claim 7 .   
     
     
         14 ) An osteoblast cell produced by a culture method as set forth in  claim 9 .