Induction of motor neurons from pluripotent stem cells
First Claim
1. A method of generating a motor neuron from a pluripotent stem cell in vitro, comprising in order from (1) to (3):
- (1) introducing a vector comprising an inducible promoter operably linked to Lhx3, Ngn2, and Isl1 genes into the pluripotent stem cell;
(2) inducing expression of the Lhx3, Ngn2, and Isl1 genes in the pluripotent stem cell from (1) in KSR media or N3 media containing a drug capable of inducing an activation of the promoter; and
(3) culturing the pluripotent stem cell from (2) and maintaining expression of the Lhx3, Ngn2, and Isl1 genes for three days or more in said cell, thereby producing a motor neuron.
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Abstract
The present invention aims to provide a method of producing motor neurons/neurons that sufficiently reproduce intrinsic properties of motor neurons/neurons, especially of the motor neurons/neurons in patients, from pluripotent stem cells promptly and synchronically, and a pluripotent stem cell capable of differentiating into a neuron or a motor neuron promptly and synchronically after a drug treatment.
A method of generating a motor neuron from a pluripotent stem cell, comprising the following steps in order from (1) to (2):
(1) introducing one or more nucleic acids encoding Lhx3, Ngn2, and Isl1 into a pluripotent stem cell; and
(2) maintaining expression of the Lhx3, Ngn2, and Isl1 for three days or more.
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Citations
7 Claims
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1. A method of generating a motor neuron from a pluripotent stem cell in vitro, comprising in order from (1) to (3):
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(1) introducing a vector comprising an inducible promoter operably linked to Lhx3, Ngn2, and Isl1 genes into the pluripotent stem cell; (2) inducing expression of the Lhx3, Ngn2, and Isl1 genes in the pluripotent stem cell from (1) in KSR media or N3 media containing a drug capable of inducing an activation of the promoter; and (3) culturing the pluripotent stem cell from (2) and maintaining expression of the Lhx3, Ngn2, and Isl1 genes for three days or more in said cell, thereby producing a motor neuron. - View Dependent Claims (2, 3, 4, 5, 6, 7)
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Specification