Novel graft polymer, temperature-responsive substrate for cell culture using the same and production method therefor, as well as liquid chromatographic carrier having the novel graft polymer immobilized thereon and liquid chromatographic method using the same

Abstract

By using a graft polymer comprising a dendritic polymer with a styrene skeleton and a hydrophilic polymer grafted to a terminal thereof, a temperature-responsive substrate for cell culture having a temperature-responsive surface for cell culture that allows cells to be cultured with high efficiency and which yet allows cultured cells to be exfoliated in a short period of time and with high efficiency by simply changing the temperature of the substrate surface can be prepared conveniently. If this temperature-responsive substrate for cell culture is used, cells obtained from a variety of tissues can be cultured with high efficiency. If this culture method is utilized, cultured cells can be exfoliated intact in a short amount of time with high efficiency. In addition, by using this graft polymer, a wide range of peptides and proteins can also be separated by simply changing the temperature of a chromatographic carrier. This allows for convenient separation procedure and improves the efficiency of separating operations. What is more, the stereoregularity of the dendritic polymer per se may be utilized to enable separation of solutes based on differences in their molecular structures.

1 . A temperature-responsive substrate for cell culture comprising a substrate coated with a dendritic polymer with a styrene skeleton and a temperature-sensitive polymer grafted at a terminal thereof. 
     
     
         2 . The temperature-responsive substrate for cell culture according to  claim 1 , which has electric charges at a terminal of the dendritic polymer. 
     
     
         3 . The temperature-responsive substrate for cell culture according to  claim 1 , wherein the polymer having temperature response comprises any one or more of a poly-N-substituted acrylamide derivative, a poly-N-substituted methacrylamide derivative, a copolymer thereof, polyvinyl methyl ether, and partially acetylated polyvinyl alcohol, or a copolymer thereof with another monomer. 
     
     
         4 . The temperature-responsive substrate for cell culture according to  claim 1 , wherein the polymer having temperature response is poly-N-isopropylacrylamide. 
     
     
         5 . The temperature-responsive substrate for cell culture according to  claim 3 , wherein the another monomer is a monomer having electric charges and/or a hydrophobic monomer. 
     
     
         6 . The temperature-responsive substrate for cell culture according to  claim 1 , wherein any one or more of polyacrylamide, poly-N,N-diethylacrylamide, poly-N,N-dimethylacrylamide, acrylate having polyethylene oxide in side chains, and methacrylate having polyethylene oxide in side chains are grafted to part of a terminal of the dendritic polymer with a styrene skeleton. 
     
     
         7 . The temperature-responsive substrate for cell culture according to  claim 1 , which has a coating of 1.0 to 7.0 μg/cm 2  in terms of the temperature-responsive polymer. 
     
     
         8 . The temperature-responsive substrate for cell culture according to  claim 1 , wherein the content of the temperature-responsive polymer in the graft polymer ranges from 40 to 99.5 wt %. 
     
     
         9 . The temperature-responsive substrate for cell culture according to  claim 1 , wherein the molecular weight of the temperature-responsive polymer in the graft polymer is 5000 or more. 
     
     
         10 . The temperature-responsive substrate for cell culture according to  claim 1 , wherein a substrate is in the form of a particle, a filament or a plate either individually or in combination of two or more. 
     
     
         11 . The temperature-responsive substrate for cell culture according to  claim 1 , wherein a substrate is composed of polystyrene either alone or combined with another material.