Neural networks formed from cells derived from pluripotent stem cells
In some aspects, cultures of neurons derived from human induced pluripotent stem cells (iPS cells) that exhibit synchronous firing of neural networks are provided. In some embodiments, neuronal activity of the cultures may be detected or measured using a multi-electrode array.
1. An in vitro method for producing a cell culture comprising a population of neurons, wherein the method comprises;
(a) combining or mixing a plurality of excitatory neurons and inhibitory neurons in vitro at a ratio of higher proportion of excitatory neurons compared to inhibitory neurons, and
b) culturing the neurons for a period of lime sufficient to allow for the formation of a neural network; wherein both the excitatory neurons and the inhibitory neurons are obtained from human induced pluripotent stem (iPS) cells; wherein the ratio of excitatory neurons to inhibitory neurons is sufficient to allow for the generation of synchronous neuron firing or synchronous action potentials by the neural network; wherein the population of neurons comprises from about 40% to about 90% excitatory neurons and from about 60% to about 10% inhibitory neurons; wherein the excitatory neurons form glutarnatergic synapses and express vGLUT1; and wherein the inhibitory neurons form GABA synapses and express vGAT and GAD67.
2. The method of claim 1 , wherein the culture of neurons comprises astrocytes, wherein the astrocytes are obtained from pluripotent stem cells.
3. The method of claim 1 , wherein the iPS cells are obtained from a healthy donor.
4. The method of claim 1 , wherein the iPS cells are obtained from a subject with a disease.
5. The method of claim 4 , wherein the disease is a neurological or neurodegenerative disease.
6. The method of claim 5 , wherein the disease is autism, epilepsy, schizophrenia, ADHD, ALS, or a bipolar disorder.
7. The method of claim 1 , wherein the population of neurons comprises from about 45% to about 80% excitatory neurons and from about 55% to about 20% inhibitory neurons.
8. The method of claims 7 , wherein from about 5% to about 25% of the cells in the cell culture are astrocytes, wherein the astrocytes are obtained from iPS cells.
9. The method of claim 8 , wherein the astrocytes are obtained from human iPS cells.
10. The method of claim 1 , wherein the population of neurons is cultured on a multi-electrode array.
11. The method of claim 10 , wherein the multi-electrode array is a Maestro MEA.
12. The method of claim 10 , wherein the multi-electrode array comprises at least 8 electrodes.
13. The method of claim 10 , wherein the multi-electrode array comprises at least 16 electrodes.
14. The method of claim 10 , wherein the multi-electrode array comprises a plurality of cultures of said neurons in distinct wells in the multi-electrode array, wherein the ratios of excitatory to inhibitory neurons varies between the cultures.
15. The method of claim 1 , wherein the method further comprises contacting the population of neurons with a test compound.
16. The method of claim 15 , wherein the test compound can modulate neurotransmission.
17. The method of claim 1 , further comprising detecting or measuring the electrical or neuronal activity of the population.
18. The method of claim 17 , wherein said detecting or measuring comprises measuring the voltage produced by said population.
19. The method of claim 18 , wherein neuronal electrical activity data from said detecting or measuring is analyzed, and wherein the population of neurons is cultured on a multi-electrode array.
20. The method of claim 1 , wherein the population of neurons is cultured on a multi-electrode array, and wherein the synchronicity is detected in a plurality of electrodes on the multi-electrode array.
21. The method of claim 7 , wherein the population of neurons comprises from about 45% to about 55% excitatory neurons and from about 55% to about 45% inhibitory neurons.
22. The method of claim 2 , wherein from about 10% to about 55% of cells in the cell culture are astrocytes, wherein the astrocytes are obtained from human iPSC.