# US 10,870,831 B2 · Method for inducing differentiation of neural stem cells using patterned hydrogel

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- Number: US 10,870,831 B2
- Document ID: US10870831B2
- Status: Granted
- Kind: B2
- Application number: 15/736,418
- Filed: 2017-03-10
- Published: 2019-02-28
- Granted: 2020-12-22
- Earliest claimed: 2016-11-11
- Assignees: Sogang University Research Foundation
- Applicants: Sogang University Research Foundation
- Inventors: Bong Geun Chung; Jong Min Lee; Joo Yoon Moon; Tae Hyeon Kim; Daniel Christian Ahrberg
- Web page: https://patentgrep.com/patent/US10870831B2

## CPC

- C12N 5/0619: {Neurons}
- C12N 5/0068: {General culture methods using substrates (for specific animal cell type C12N5/06)}
- C12N 5/0623: {Stem cells}
- C12N 13/00: Treatment of microorganisms or enzymes with electrical or wave energy, e.g. magnetism, sonic waves
- C12N 2500/10: Metals; Metal chelators
- C12N 2500/50: Soluble polymers, e.g. polyethyleneglycol [PEG]
- C12N 2529/00: Culture process characterised by the use of electromagnetic stimulation
- C12N 2533/30: Synthetic polymers
- C12N 2535/00: Supports or coatings for cell culture characterised by topography
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)

## Abstract

The present invention provides a method for inducing differentiation of neural stem cells. The present invention provides optimized differentiation conditions of neural stem cells into neurons using a patterned hydrogel.

## Claims

### Claim 1

A method for inducing differentiation of neural stem cells, the method comprising:
(a) culturing neural stem cells to induce neurospheres;
(b) seeding the neurospheres on a cell culture substrate on which a hydrogel comprising a biocompatible polymer and a conductive nanowire is patterned to have intervals of 150-250 μm; and
(c) applying voltage to the cell culture substrate to induce the differentiation into neurons,
wherein the conductive nanowires are patterned by mixing with the biocompatible polymer, and fixed by photocrosslinking of the biocompatible polymer, and
wherein the hydrogel in step (b) comprises a biocompatible polymer and a conductive nanowire at a mixing ratio of 8-12:1.

### Claim 2 (depends on 1)

The method of claim 1, wherein the neurospheres in step (a) have a size of 100-150 μm.

### Claim 3 (depends on 1)

The method of claim 1, wherein the biocompatible polymer in step (b) is polyethylene glycol (PEG).

### Claim 4 (depends on 1)

The method of claim 1, wherein the conductive nanowire in step (b) is a metal nanowire.

### Claim 5 (depends on 4)

The method of claim 4, wherein the metal nanowire is a silver nanowire, a copper nanowire, a gold nanowire, or an aluminum nanowire.

### Claim 6 (depends on 1)

The method of claim 1, wherein the voltage in step (c) is 8-12 V.

### Claim 7 (depends on 1)

The method of claim 1, wherein the cell culture substrate in step (c) has an average current value of 10-20 nA.