Method for sorting tissue cells

Abstract

An object of the present invention is to provide a method for increasing the purity of a type of tissue cell such as an endothelial cell, a hepatocyte, or an insulin-producing cell. The present invention solves the problem by providing a method comprising a step of introducing, into a cell population, an mRNA comprising a nucleic acid sequence recognized by an miRNA specifically expressed in endothelial cells, hepatocytes, or insulin-producing cells.

1 - 11 . (canceled) 
     
     
         12 . A method for producing hepatocytes, comprising:
 (a) producing, from a pluripotent stem cell, a cell population containing hepatocytes;   (b) introducing, into the cell population of the step (a), an miRNA-responsive mRNA consisting of sequences comprising (i) a nucleic acid recognized specifically by miR-122-5P and (ii) a nucleic acid corresponding to a coding region of a first marker gene, wherein the miRNA-responsive mRNA is introduced into the cell population in a form of a synthetic RNA molecule; and   (c) sorting a cell having a low level of translation of the first marker gene of the step (b).   
     
     
         13 . (canceled) 
     
     
         14 . The method according to  claim 12 , wherein the miRNA-responsive mRNA comprises a nucleic acid sequence having (i) and (ii) linked in a 5′ to 3′ direction. 
     
     
         15 . The method according to  claim 12 , wherein the first marker gene of (ii) is one or more genes selected from the group consisting of a fluorescent protein-encoding gene, an apoptosis-inducing gene, and a suicide gene. 
     
     
         16 . The method according to  claim 12 , further comprising introducing, into the cell population, an mRNA consisting of a sequence comprising a nucleic acid corresponding to a coding region of a second marker gene, wherein a cell in which the second marker gene is translated is sorted as the cell of interest, and wherein the second marker gene is a fluorescent protein-encoding gene different from the first marker gene, or a drug resistance gene.