Production of primate neural stem cells through expression of pax6
A transcription factor both necessary and sufficient for human neuroectoderm specification, Pax6, as well as applications thereof, is disclosed.
1. A method of creating a population of primate pNSCs from primate embryonic stem cells (ESCs) or induced pluripotent stem cells (iPSCs) comprising the step of:
introducing a nucleic acid sequence encoding Pax6 into primate ESCs or iPSCs, wherein Pax6 production within the cells is sufficient produce Pax6+Sox1-primate pNSCs.
2. The method of claim 1 , wherein the introduction of Pax6 is via an inducible system.
3. The method of claim 1 , wherein the introduction of Pax6 is via a lentiviral vector.
4. The method of claim 1 , wherein the introduction of Pax6 is under the control of elongation factor Ia promoter in the lentiviral vector.
5. The method of claim 3 , wherein the lentiviral vector is an inducible lentiviral vector.
6. The method of claim 1 , wherein the primate is human.
7. A method of creating a population of primate regional neural stem cells comprising the steps of:
(a) introducing a nucleic acid sequence encoding Pax6 into primate ESCs or iPSCs, wherein Pax6 production within the cells is sufficient produce Pax6+Sox1− primate pNSCs;
(b) suppressing Pax6 production; and
(c) differentiating the cells of step (b) into regional neural stem cells.
8. The method of claim 7 wherein the regional neural stem cells are selected from the group consisting of forebrain cells, midbrain cells and spinal cells.
9. The method of claim 7 wherein the primate is human.
10. A method of creating a population of primate pNSCs from primate regional neural stem cells comprising the step of:
introducing a nucleic acid sequence encoding Pax6 into isolated primate regional neural stem or progenitor cells, wherein Pax6 production within the cells is sufficient produce Pax6+Sox1− primate pNSCs is sufficient to reprogram the cells to the primate pNSC stage.
11. The method of claim 10 wherein the primate is human.