Direct conversion of cells to cells of other lineages

Abstract

Methods, compositions and kits for producing functional neurons, astroctyes, oligodendrocytes and progenitor cells thereof are provided. These methods, compositions and kits find use in producing neurons, astrocytes, oligodendrocytes, and progenitor cells thereof for transplantation, for experimental evaluation, as a source of lineage- and cell-specific products, and the like, for example for use in treating human disorders of the CNS. Also provided are methods, compositions and kits for screening candidate agents for activity in converting cells into neuronal cells, astrocytes, oligodendrocytes, and progenitor cells thereof.

That which is claimed is: 
     
         1 . A method of converting pluripotent cells into induced neuronal cells, the method comprising:
 contacting a population of pluripotent cells with an effective dose of a neuron reprogramming (NR) system comprising one or more neuron reprogramming (NR) factors, wherein the NR factors are selected from the group consisting of: an Ascl agent, a Ngn agent, and a NeuroD agent for a period of time sufficient to reprogram said pluripotent cells, wherein a population of induced neuronal cells is produced.   
     
     
         2 . The method of  claim 1 , wherein the pluripotent cells are human cells. 
     
     
         3 . The method of  claim 1 , wherein the efficiency of reprogramming said pluripotent cells to become induced neuronal cells is at least about 0.1%. 
     
     
         4 . The method of  claim 1 , wherein the pluripotent cells are reprogrammed into neuronal cells with functional properties as early as 10 days after contacting the cells with the reprogramming factors. 
     
     
         5 . The method of  claim 1 , wherein the pluripotent cells are induced pluripotent (iPS) cells. 
     
     
         6 . The method of  claim 1 , wherein the pluripotent cells are embryonic stem cells. 
     
     
         7 . The method of  claim 1 , wherein the neuron reprogramming factor is Ngn2. 
     
     
         8 . The method of  claim 1 , wherein the induced neuronal cells are excitatory neurons. 
     
     
         9 . A cell culture system comprising a population of pluripotent cells with an effective dose of a neuron reprogramming (NR) system comprising one or more neuron reprogramming (NR) factors, wherein the NR factors are selected from the group consisting of: an Ascl agent, a Ngn agent, and a NeuroD agent. 
     
     
         10 . The cell culture system of  claim 9 , wherein the pluripotent cells are induced pluripotent (iPS) cells. 
     
     
         11 . The cell culture system of  claim 9 , wherein the pluripotent cells are embryonic stem cells. 
     
     
         12 . The cell culture system of  claim 9 , wherein the neuron reprogramming factor is Ngn2. 
     
     
         13 . A method of determining the effect of a candidate agent on a neuron output parameter, the method comprising:
 contacting a population of induced neuronal cells produced by the method of  claim 1  with a candidate agent; and   determining the effect on an output parameter.   
     
     
         14 . The method of  claim 13 , wherein the output parameter is selected from induced neuronal cell survival, the ability of induced neuronal cells to become depolarized, the extent to which the induced neuronal cells form synapses. 
     
     
         15 . A method of converting human pluripotent cells into induced neuronal cells, the method comprising:
 contacting a population of human pluripotent cells with an effective dose of a neuron reprogramming (NR) system comprising Ngn2 as a reprogramming factor for a period of time sufficient to reprogram said pluripotent cells,   wherein a population of induced neuronal cells is produced.   
     
     
         16 . The method of  claim 15 , wherein the efficiency of reprogramming said pluripotent cells to become induced neuronal cells is at least about 0.1%. 
     
     
         17 . The method of  claim 15 , wherein the pluripotent cells are reprogrammed into neuronal cells with functional properties within 10 days after contacting the cells with the reprogramming factors. 
     
     
         18 . The method of  claim 15 , wherein the pluripotent cells are induced pluripotent (iPS) cells. 
     
     
         19 . The method of  claim 15 , wherein the pluripotent cells are embryonic stem cells. 
     
     
         20 . The method of  claim 15 , wherein the induced neuronal cells are excitatory neurons.