Microvesicles derived from nucleated, mammalian cells and use thereof

Abstract

The present invention relates to a microvesicle that is derived from nucleated mammalian cells, which are smaller than the nucleated cells. The microvesicles of the present invention can be used in the delivery of a therapeutic or diagnostic substance to specific tissues or cells, and more particularly, relates to microvesicles derived from monocytes, macrophages, dendritic cells, stem cells or the like, which can be used to deliver specific therapeutic or diagnostic substances for treating and/or diagnosing tissue associated with cancer, diseased blood vessels, inflammation, or the like.

1 . A method of producing sub-cell-sized vesicles, the method comprising:
 providing nucleated mammalian cells having a cell membrane topology, in which at least one membrane protein has an extracellular domain located outside membrane of the nucleated mammalian cells;   extruding the nucleated mammalian cells to produce sub-cell-sized vesicles maintaining the cell membrane topology such that the extracellular domain of the at least one membrane-protein is located outside membrane of the vesicles; and   to collecting at least part of the produced vesicles.   
     
     
         2 . The method of  claim 1 , further comprising:
 mixing the collected vesicles in the presence of a therapeutic or diagnostic substance to load the therapeutic or diagnostic substance in the vesicles.   
     
     
         3 . A method of treating or diagnosing a disease, the method comprising:
 the method of  claim 2  to provide the vesicles loaded with the therapeutic or diagnostic substance;   processing the loaded vesicles to provide a composition comprising at least part of the loaded vesicles; and   administering the composition to a subject in need of such a treatment or diagnosis.   
     
     
         4 . The method of  claim 1 , further comprising:
 prior to extruding, mixing the nucleated mammalian cells with a medium comprising a therapeutic or diagnostic substance; and   wherein at least part of the collected produced vesicles contain the therapeutic or diagnostic substance.   
     
     
         5 . A method of treating or diagnosing a disease, the method comprising:
 the method of  claim 4  to provide the collected vesicles;   processing the collected vesicles to provide a composition comprising at least part of the collected vesicles; and   administering the composition to a subject in need of such a treatment or diagnosis.   
     
     
         6 . A method of treating or diagnosing a disease, the method comprising:
 providing the vesicles loaded with the therapeutic or diagnostic substance in accordance with  claim 2 ;   processing the loaded vesicles to provide a composition comprising at least part of the loaded vesicles; and   administering the composition to a subject in need of such a treatment or diagnosis.   
     
     
         7 . A method of producing sub-cell-sized vesicles, the method comprising:
 providing nucleated mammalian cells having a cell membrane topology, in which at least one membrane protein has an extracellular domain located outside membrane of the nucleated mammalian cells;   processing the nucleated mammalian cells to artificially produce sub-cell-sized vesicles, substantially all of which maintain the cell membrane topology such that the extracellular domain of the at least one membrane protein is located outside membrane of the vesicles; and   collecting at least part of the produced vesicles.   
     
     
         8 . The method of  claim 7 , wherein processing the nucleated mammalian cells to artificially produce sub-cell-sized vesicles is selected from the group consisting of extrusion, homogenization, freezing-thawing, electroporation, and mechanical degradation. 
     
     
         9 . The method of  claim 7 , further comprising:
 mixing the collected vesicles in the presence of a therapeutic or diagnostic substance to load the therapeutic or diagnostic substance in the vesicles.   
     
     
         10 . A method of treating or diagnosing a disease, the method comprising:
 the method of  claim 9  to provide the vesicles loaded with the therapeutic or diagnostic substance;   processing the loaded vesicles to provide a composition comprising at least part of the loaded vesicles; and   administering the composition to a subject in need of such a treatment or diagnosis.   
     
     
         11 . The method of  claim 7 , further comprising:
 prior to processing, mixing the nucleated mammalian cells with a medium comprising a therapeutic or diagnostic substance; and   wherein at least part of collected the produced vesicles contain the therapeutic or diagnostic substance.   
     
     
         12 . A method of treating or diagnosing a disease, the method comprising:
 the method of  claim 11  to provide the collected vesicles;   processing the collected vesicles to provide a composition comprising at least part of the collected vesicles; and   administering the composition to a subject in need of such a treatment or diagnosis.