Reprogrammation of Eukaryotic Cells with Engineered Microvesicles

Abstract

The present invention relates to a non-genetic, detergent-free, bacteria-free method for reprogramming a eukaryotic cell, in particular for obtaining induced pluripotent stem cells (iPS), by using engineered microvesicles carrying at least one reprogramming transcription factor, wherein said engineered microvesicles are virus-free.

1 - 14 . (canceled) 
     
     
         15 . An isolated microvesicle as released from a eukaryotic cell, wherein said microvesicle comprises a viral membrane fusion protein and a reprogramming transcription factor or ecotropic receptor. 
     
     
         16 . The isolated microvesicle of claim  1 , wherein said microvesicle does not comprise any nucleic acid coding for said reprogramming transcription factor or ecotropic receptor. 
     
     
         17 . The isolated microvesicle of claim  1 , wherein said microvesicle does not contain any viral structural protein. 
     
     
         18 . The isolated microvesicle of claim  1 , which is an exosome-like vesicle having a size between 40 and 150 nm. 
     
     
         19 . The isolated microvesicle of claim  1 , which is an exosome-like vesicle having a density between 1.08 g/ml and 1.11 g/ml. 
     
     
         20 . The isolated microvesicle of claim  1 , obtained by the filtration and ultracentrifugation of cell supernatants of eukaryotic cells, wherein said eukaryotic cells overexpress said viral membrane fusion protein and said reprogramming transcription factor, and wherein said eukaryotic cell does not express any viral structural proteins. 
     
     
         21 . The isolated microvesicle of claim  1 , wherein said viral membrane fusion protein is VSV-G or baculovirus gp64. 
     
     
         22 . The isolated microvesicle of claim  1 , wherein said reprogramming transcription factor is selected from the group consisting of Oct-4, KLF4, sox2, Lin28 and c-myc. 
     
     
         23 . A kit for delivering an EcoR ecotropic viral receptor to a target cell, comprising:
 a. the isolated microvesicle of claim  1 , wherein said ecotropic viral receptor is EcoR ecotropic viral receptor.   
     
     
         24 . A kit for obtaining an iPS form a target cell, comprising:
 a. the isolated microvesicle of claim  1 , with Oct4 as the reprogramming transcription factor;   b. the isolated microvesicle of claim  1 , with KLF-4 as the reprogramming transcription factor;   c. the isolated microvesicle of claim  1 , with sox2 as the reprogramming transcription factor; and   d. the isolated microvesicle of claim  1 , with a reprogramming transcription factor selected from the group consisting of Lin28 and c-myc.