Implantable living electrodes and methods for use thereof
In one aspect, the invention comprises an implantable living electrode comprising a substantially cylindrical extracellular matrix core; one or more neurons implanted along or within the substantially cylindrical extracellular matrix core, the one or more neurons including one or more optogenetic or magnetogenetic neurons proximal to a first end of the implantable living electrode.
1. A method of manufacturing an implantable living electrode comprising:
(a) providing an extracellular matrix core of a micro-column;
(b) centrifuging a solution of neurons within one or more micro-wells to produce, via forced aggregation, one or more preformed neuronal aggregates, each comprising a plurality of neurons aggregated together; and
(c) contacting at least one end of the extracellular matrix core with the one or more preformed neuronal aggregates.
2. The method according to claim 1 , further comprising maintaining the implantable living electrode under conditions that promote axon growth within or along the extracellular matrix core.
3. The method of claim 1 , wherein each of the one or more preformed neuronal aggregates is a spheroidal aggregate.
4. The method of claim 1 , wherein the one or more micro wells comprise inverted pyramidal micro-wells.
5. The method of claim 1 , wherein a density of neurons in the solution of neurons is at least 1 million cells/mL.
6. The method of claim 1 , further comprising:
(d) culturing the micro-column and extracellular matrix core with the one or more preformed neuronal aggregates therein for one or more days in vitro to promote growth of axonal projections from neurons within the one or more preformed neuronal aggregates into the cylindrical extracellular matrix core, such that aggregated neuronal soma occupy a zone at one or both ends of the micro-column and axonal projections of the neurons run longitudinally along at least a portion of the micro-column, directed from ends and towards a center of the micro-column.
7. The method of claim 1 , wherein the one or more preformed neuronal aggregates comprise dopaminergic neurons.
8. The method of claim 7 , wherein the dopaminergic neurons comprise dopaminergic A9 neurons.
9. The method of claim 1 , wherein step (a) comprises 3D printing the micro-column.
10. A method of manufacturing an implantable living electrode comprising:
(a) providing an extracellular matrix core of a micro-column; and
(b) contacting at least one end of the extracellular matrix core with one or more spheroidal aggregates, each (i) comprising a plurality of aggregated neurons and (ii) implanted at an end of, or within, the extracellular matrix core.
11. The method of claim 10 , comprising, prior to step (b), forming, via forced aggregation, the one or more spheroidal aggregates from a solution of neurons.
12. The method of claim 11 , wherein forming the one or more spheroidal aggregates comprises:
transferring, into each of one or more micro-wells, a dissociated neuron suspension comprising a plurality of dissociated neurons; and
centrifuging the one or more micro-wells with the dissociated neuron suspension therein to produce the one or more spheroidal aggregates.
13. The method of claim 12 , wherein a density of neurons in the dissociated neuron suspension is at least 1 million cells/mL.
14. The method according to claim 10 , further comprising maintaining the implantable living electrode under conditions that promote axon growth within or along the extracellular matrix core.
15. The method of claim 10 , further comprising:
(c) culturing the micro-column and extracellular matrix core with the one or more spheroidal aggregates therein for one or more days in vitro to promote growth of axonal projections from neurons within the one or more spheroidal aggregates into the cylindrical extracellular matrix core, such that aggregated neuronal soma occupy a zone at one or both ends of the micro-column and axonal projections of the neurons run longitudinally along at least a portion of the micro-column, directed from ends and towards a center of the micro-column.
16. The method of claim 10 , wherein the one or more spheroidal aggregates comprise dopaminergic neurons.
17. The method of claim 16 , wherein the dopaminergic neurons comprise dopaminergic A9 neurons.
18. The method of claim 10 , wherein step (a) comprises 3D printing the micro-column.