Generation of neural stem cells and motor neurons
A method of generating neural stem cells or motor neurons is disclosed, the method comprising up-regulating a level of at least one exogenous miRNA and/or down-regulating at least one miRNA using an agent which hybridizes to the miRNA in mesenchymal stem cells (MSCs) or down-regulating Related to testis-specific, vespid and pathogenesis protein 1 (RTVP-1).
1. A method of promoting mesenchymal stem cell (MSC) differentiation toward a neuronal stem cell, the method comprising:
(i) culturing a MSC in a medium supporting neuronal stem cell growth and differentiation,
(ii) introducing into said MSC at least one exogenous microRNA (miR) selected from the group consisting of miR-20B, miR-378 and miR-891,
(iii) introducing into said MSC a miR-138 antagomir, and
(iv) confirming increased expression at least one neuronal marker selected from the group consisting of nesting and Sox2 by detecting expression of said marker on said MSC, thereby promoting differentiation of the MSC into the neuronal stem cell.
2. The method of claim 1 , wherein said introducing comprises any one of:
(i) transfecting said MSCs with an expression vector which comprises a polynucleotide sequence which encodes a pre-miRNA of said miR; or
(ii) transfecting said MSCs with an expression vector which comprises a polynucleotide sequence which encodes said miR.
3. A method of promoting mesenchymal stem cell (MSC) differentiation toward a neuronal stem cell, the method comprising:
(i) culturing a MSC in a medium supporting neuronal stem cell growth and differentiation,
(ii) introducing into said MSC at least one exogenous microRNA (miR) selected from the group consisting of miR-20B, miR-378 and miR-891,
(iii) introducing into said MSC a miR-138 antagomir,
(iv) confirming expression at least one neuronal marker selected from the group consisting of nesting and Sox2, wherein said expressing results in at least 50% of the MSCs expressing said neuronal marker, thereby promoting differentiation of the MSC into the neuronal stem cell.
4. The method of claim 3 , wherein said introducing comprises any one of:
(i) transfecting said MSCs with an expression vector which comprises a polynucleotide sequence which encodes a pre-miRNA of said miR; or
(ii) transfecting said MSCs with an expression vector which comprises a polynucleotide sequence which encodes said miR.