Micropillar arrays for assaying myelination
View Patent ↗Micropillar arrays for assaying differentiation of oligodendrocyte precursor cells into oligodendrocytes, ensheathment, and/or wrapping of the micropillars by the oligodendrocytes is provided. Also provided herein are methods of using the micropillar arrays for screening of candidate agents that promote differentiation of oligodendrocyte precursor cells into oligodendrocytes, ensheathment, and/or wrapping of the micropillars by the oligodendrocytes. A system comprising micropillar arrays and oligodendrocyte precursor cells are also provided.
1. A method for screening for candidate agents that promote myelination, the method comprising:
incubating a candidate agent with a substrate comprising a plurality of micropillars and oligodendrocyte precursor cells (OPCs); and
determining presence or absence of myelination on the plurality of micropillars by visualization of myelin rings around a circumference of the micropillars,
wherein the presence of myelination of the plurality of micropillars indicates that the candidate agent promotes myelination,
wherein the plurality of micropillars comprise micropillars that project vertically from the substrate and comprise a shape that tapers in diameter from a base of the micropillars attached to the substrate to a tip of the micropillars projecting away from the substrate.
2. The method of claim 1 , wherein the determining presence or absence of myelination comprises assaying for differentiation of OPCs into oligodendrocytes, wherein the assaying comprises detecting ensheathment of the plurality of micropillars by the oligodendrocytes.
3. The method of claim 1 , wherein the determining presence or absence of myelination comprises assaying for concentric wrapping of the plurality of micropillars by oligodendrocytes generated from the OPCs.
4. The method of claim 1 , wherein the determining presence or absence of myelination comprises determining a level of myelination.
5. The method of claim 1 , wherein the micropillars comprise a base ranging in diameter from 10 μm to 150 μm and a tip ranging in diameter from 0.1 μm to 10 μm.
6. The method of claim 1 , wherein the micropillars comprise a height between the tip and the base ranging from 15 μm to 50 μm.
7. The method of claim 1 , wherein the shape comprises a conical shape, frusto-conical shape, a bi-conical shape, or a parabolic shape.
8. The method of claim 1 , wherein the base ranges in diameter from 25 μm to 50 μm, the tip ranges in diameter from 1 μm to 5 μm, and the height between the tip and the base ranges from 25 μm to 50 μm.
9. The method of claim 1 , wherein the shape comprises a conical shape.
10. The method of claim 1 , wherein the substrate comprises a plurality of chambers and wherein the plurality of chambers each comprise a plurality of micropillars and the OPCs.
11. The method of claim 10 , wherein the incubating comprises incubating a plurality of candidate agents with the substrate comprising the plurality of chambers and the OPCs, wherein the plurality of chambers each comprise a different candidate agent.
12. The method of claim 10 , wherein the plurality of chambers each comprise 10 to 200 of the micropillars.
13. The method of claim 10 , wherein the shape comprises a conical shape.
14. The method of claim 13 , wherein the base ranges in diameter from 25 μm to 50 μm, the tip ranges in diameter from 1 μm to 5 μm, and the height between the tip and the base ranges from 25 μm to 50 μm.