Method for evaluating quality of human mesenchymal stem cell, and monoclonal antibody for use in said method

Abstract

The present invention aims to obtain a method for quality evaluation of human mesenchymal stem cells, a method for isolation, selection and culture of human mesenchymal stem cells, a cell population of rapidly proliferating human mesenchymal stem cells, as well as monoclonal antibodies that specifically recognize rapidly proliferating human mesenchymal stem cells. From a cell population containing human mesenchymal stem cells, rapidly proliferating human mesenchymal stem cells are isolated, selected and cultured. The abundance ratio of cells expressing Ror2 or Fzd5 in the cell population thus isolated, selected and cultured is quantified to determine whether or not each cell population is acceptable.

1 - 20 . (canceled) 
     
     
         21 . A method for quality evaluation of human mesenchymal stem cells, which comprises:
 a step where from a cell population containing human mesenchymal stem cells, rapidly proliferating human mesenchymal stem cells are isolated, selected and cultured; and   a step where the abundance ratio of cells expressing Ror2 in the cell population thus isolated, selected and cultured is quantified to determine whether or not each cell population is acceptable.   
     
     
         22 . The method for quality evaluation of human mesenchymal stem cells according to  claim 21 , wherein the cells expressing Ror2 are quantified by using anti-Ror2 monoclonal antibody. 
     
     
         23 . The method for quality evaluation of human mesenchymal stem cells according to  claim 22 , wherein the cells expressing mRNA of Ror2 are quantified by quantitative PCR. 
     
     
         24 . The method for quality evaluation of human mesenchymal stem cells according to  claim 22 , wherein the cells expressing Ror2 are quantified by immunostaining. 
     
     
         25 . A method for isolation, selection and culture of human mesenchymal stem cells, which comprises:
 a step where from a cell population containing human mesenchymal stem cells, rapidly proliferating human mesenchymal stem cells are isolated, selected and cultured; and   a step where the abundance ratio of cells expressing Ror2 in the cell population thus isolated, selected and cultured is quantified to determine whether or not each cell population is acceptable, thus selecting only the cell population(s) determined to be acceptable.   
     
     
         26 . The method for isolation, selection and culture of human mesenchymal stem cells according to  claim 25 , wherein the cells expressing Ror2 are quantified by using anti-Ror2 monoclonal antibody. 
     
     
         27 . The method for isolation, selection and culture of human mesenchymal stem cells according to  claim 26 , wherein the cells expressing mRNA of Ror2 are quantified by quantitative PCR. 
     
     
         28 . The method for isolation, selection and culture of human mesenchymal stem cells according to  claim 25 , wherein the step where rapidly proliferating human mesenchymal stem cells are isolated, selected and cultured comprises a step where the cell population containing human mesenchymal stem cells is analyzed by flow cytometry (hereinafter referred to as “FCM”) for cells stained simultaneously with anti-LNGFR monoclonal antibody and anti-Thy1 monoclonal antibody to thereby effect cell sorting of LNGFR +  Thy1 +  cells. 
     
     
         29 . The method for isolation, selection and culture of human mesenchymal stem cells according to  claim 25 , wherein the step where rapidly proliferating human mesenchymal stem cells are isolated, selected and cultured comprises a step where the cell population containing human mesenchymal stem cells is analyzed by FCM for cells stained with anti-Ror2 monoclonal antibody to thereby effect cell sorting of Ror2 +  cells. 
     
     
         30 . The method for isolation, selection and culture of human mesenchymal stem cells according to  claim 28  or  29 , which comprises a step where the cell population is directly prepared from cells derived from bone marrow or other tissues. 
     
     
         31 . The method for isolation, selection and culture of human mesenchymal stem cells according to  claim 30 , which comprises a step where the cell population is prepared by adherent culture of cells derived from bone marrow or other tissues. 
     
     
         32 . The method for isolation, selection and culture of human mesenchymal stem cells according to  claim 28  or  29 , wherein the cell sorting step comprises a step where the positive cells are seeded in wells of a culture plate and the cells in each well reaching confluence upon culture are then isolated and selected. 
     
     
         33 . A cell population of rapidly proliferating human mesenchymal stem cells, which is obtained by:
 a step where a cell population containing human mesenchymal stem cells is analyzed by FCM for cells stained with anti-Ror2 monoclonal antibody to thereby effect cell sorting of Ror2 +  cells, and these positive cells are seeded in wells of a culture plate and the cells in each well reaching confluence upon culture are isolated and selected; and   a step where the abundance ratio of cells expressing Ror2 in the cell population thus isolated and selected is quantified to determine whether or not each cell population is acceptable, thus selecting only the cell population(s) determined to be acceptable.   
     
     
         34 . A cell population of rapidly proliferating human mesenchymal stem cells, which is obtained by:
 a step where a cell population containing human mesenchymal stem cells is analyzed by FCM for cells stained with anti-Ror2 monoclonal antibody to thereby effect cell sorting of Ror2 +  cells, and these positive cells are seeded in wells of a culture plate and the cells in each well reaching confluence upon culture are isolated and selected; and   a step where the abundance ratio of cells expressing Fzd5 in the cell population thus isolated and selected is quantified to determine whether or not each cell population is acceptable, thus selecting only the cell population(s) determined to be acceptable.   
     
     
         35 . The cell population of rapidly proliferating human mesenchymal stem cells according to  claim 33  or  34 , wherein cells whose abundance ratio is 63% or higher are determined to be acceptable.