Methods relating to pluripotent cells

Abstract

The technology described herein relates to methods, assays, and compositions relating to causing a cell to assume a more pluripotent state, e.g. without introducing foreign genetic material.

We claim: 
     
         1 . Isolated cells expressing one or more markers of pluripotency formed by treating non-embryonic, non-transformed normal differentiated mammalian somatic cells with an effective amount of chemical stress comprising ATP to a concentration of between 20 μM and 200 mM, alone or in combination with a mechanical stress to increase the levels of stress inducible genes in an amount and for a time effective to increase the number of cells expressing one or more markers of pluripotency, wherein the cells are at a pH of between 3.0 and 6.8,
 without introduction of an exogenous gene, a transcript, a protein, a nuclear component or cytoplasm, or without cell fusion, 
 wherein the one or more pluripotency markers are selected from the group consisting of Oct4, SSEA, Nanog and Sox2, thereby increasing the number of cells expressing the markers of pluripotency, 
 wherein the cells are in tissue specific cell culture media. 
 
     
     
         2 . The cells of  claim 1 , wherein the cell population is formed by the method further comprising exposure to mechanical stress disrupting pores resulting in loss of between about 40% and about 90% of the cytoplasm or the mitochondria from the cell, in addition to chemical stress. 
     
     
         3 . The cells of  claim 1 , wherein the mechanical disruption is achieved by trituration of the cells. 
     
     
         4 . The cells of  claim 1 , wherein the stress is mechanical disruption of the pores of the cell membrane in the presence of ATP and exposure to a pH between 4.6 and 6. 
     
     
         5 . The cells of  claim 4  wherein the pH is between 5 and 5.7. 
     
     
         6 . The cells of  claim 1  wherein the ATP is in a concentration between one and 15 mM. 
     
     
         7 . The cells of  claim 1  wherein the cells are cultured in culture media for culturing embryonic neural stem cells. 
     
     
         8 . The cells of  claim 1  wherein the cells are cultured in culture media for culturing skin cells. 
     
     
         9 . The cells of  claim 1  wherein the cells are cultured in culture media for culturing liver cells. 
     
     
         10 . The cells of  claim 1  wherein the cells are cultured in culture media for culturing muscle cells. 
     
     
         11 . The cells of  claim 1  wherein the cells are cultured in culture media for culturing lung cells. 
     
     
         12 . The cells of  claim 1  wherein the cells are cultured in culture media for culturing mesoderm cells.