Method for evaluation of grafts

Abstract

A method and system for evaluating the competence for transplantation of a graft sample containing cells capable of multinucleation are provided. In one form, the method includes determining multinucleating ability as an indicator representing the capacity for multinucleation of the cells capable of multinucleation.

1 - 9 . (canceled) 
     
     
         10 . A method for identifying sheet-shaped graft samples that are competent for transplantation, the method comprising:
 culturing cells of a mammal of interest to form sheet-shaped graft samples;   detecting a predetermined characteristic of cells from the sheet-shaped graft samples or a portion thereof that relate to the multinucleating ability of the cells; and   selecting the graft samples for transplantation that have a predetermined level of multinucleating ability based on the detected predetermined characteristic.   
     
     
         11 . The method of  claim 10 , further comprising treating the sheet-shaped graft samples or a portion thereof prior to detecting the predetermined characteristic of the cells to allow the predetermined characteristic of the cells to be detected. 
     
     
         12 . The method of  claim 10 , further comprising dissociating individual cells from the sheet-shaped graft samples or a portion thereof, and incubating the dissociated individual cells prior to detecting the predetermined characteristic of the cells to allow the predetermined characteristic of the cells to be detected. 
     
     
         13 . The method of  claim 10 , wherein the sheet-shaped graft samples comprise skeletal myoblasts. 
     
     
         14 . The method of  claim 10 , further comprising dissociating cells from the sheet-shaped graft samples or a portion thereof and incubating the dissociated cells to detect the predetermined characteristic thereof. 
     
     
         15 . The method of  claim 10 , wherein detecting the predetermined characteristic of cells from the sheet-shaped graft samples or a portion thereof comprises identifying multinucleate cells by at least one of visual observation and imaging. 
     
     
         16 . The method of  claim 15 , further comprising staining nuclei of the cells with a dye or fluorescent substance prior to identifying multinucleate cells by the at least one of visual observation and imaging. 
     
     
         17 . The method of  claim 10 , wherein detecting the predetermined characteristic of cells comprises detecting at least one of:
 the number of multinucleate cells existing in the sheet-shaped graft samples or a portion thereof;   the rate of multinucleation in the sheet-shaped graft sample or a portion thereof;   the number of all nuclei existing in multinucleate cells in the sheet-shaped graft sample or a portion thereof;   the amount of cytokine yielded by the sheet-shaped graft sample or a portion thereof; and   the amount of proteins in the sheet-shaped graft sample or a portion thereof.   
     
     
         18 . A method for identifying graft samples that are competent for transplantation, the method comprising:
 culturing cells of a mammal of interest to form sheet-shaped graft samples;   obtaining individual cells from at least a portion of the sheet-shaped graft samples;   exposing the individual cells to predetermined conditions to encourage multinucleation; and   detecting a predetermined characteristic of the cells related to the multinucleating ability of the cells in the sheet-shaped graft samples.   
     
     
         19 . The method of  claim 18 ; wherein the individual cells are obtained by dissociating at least a portion of at least one sheet shaped graft sample into individual cells. 
     
     
         20 . The method of  claim 18 , further comprising inoculating the individual cells onto a culture medium and incubating the individual cells. 
     
     
         21 . The method of  claim 18 , wherein detecting the predetermined characteristic of the individual cells comprises measuring the number of multinucleate cells. 
     
     
         22 . The method of  claim 21 , wherein measuring the number of multinucleate cells comprises staining nuclei of the individual cells with a dye or fluorescent substance and identifying multinucleate cells by at least one of visual observation and imaging. 
     
     
         23 . The method of  claim 18 , wherein detecting the predetermined characteristic of the cells comprises staining the nuclei with a fluorescent substance and sorting the cells having a fluorescent intensity that is at or above a predetermined level. 
     
     
         24 . The method of  claim 18 , wherein detecting the predetermined characteristic of the cells related to the multinucleating ability comprises comparing the detected predetermined characteristic against at least one preset threshold value for the predetermined characteristic. 
     
     
         25 . The method of  claim 18 , wherein detecting the predetermined characteristic of the cells related to the multinucleating ability comprises at least one of determining at least one of the number of multinucleate cells, the rate of multinucleation, and the number of nuclei resulting from multinucleation. 
     
     
         26 . The method of  claim 18 , wherein the sheet-shaped graft samples comprise skeletal myoblasts.