# US 10,871,435 B2 · Methods and kits for cell sorting

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- Number: US 10,871,435 B2
- Document ID: US10871435B2
- Status: Granted
- Kind: B2
- Application number: 15/652,928
- Filed: 2017-07-18
- Published: 2019-01-24
- Granted: 2020-12-22
- Earliest claimed: 2017-07-18
- Applicants: Shengyuan Yang
- Inventors: Shengyuan Yang
- Web page: https://patentgrep.com/patent/US10871435B2

## CPC

- G01N 15/10: Investigating individual particles
- C12M 3/00: Tissue, human, animal or plant cell, or virus culture apparatus
- C12M 25/16: {Particles; Beads; Granular material; Encapsulation (chemical or physical processes conducted in the presence of fluids and solid particles B01J8/00)}
- C12M 47/04: {Cell isolation or sorting (purging biological preparations of unwanted cells C12N5/0081, determining the presence or kind of microorganism C12Q1/04)}
- G01N 33/4833: {of solid biological material, e.g. tissue samples, cell cultures (tissue in vivo A61B5/00; cell suspensions G01N33/48735)}
- G01N 15/01: specially adapted for biological cells, e.g. blood cells (investigating sedimentation of particle suspensions in blood G01N15/05)
- G01N 2015/1006: {for cytology}
- G01N 2015/1028: {Sorting particles}
- G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q)
- C12M: APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; {APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS}

## Abstract

The present disclosure provides methods, and related kits, for directing cell attachment and spreading on a substrate and inducing isotropic spreading of cells; provides methods, and related kits, for cell sorting; and further provides methods, and related kits, for guided induction of stem cell differentiation.

## Claims

### Claim 1

A method of cell sorting based on the different adherent sensitivities to substrate curvatures of different cell types, comprising:
culturing a mixed population of fibroblasts and mesenchymal stem cells on a glass spherical substrate having a diameter of 58-500 μm in cell culture media; and
removing or separating the cell culture media from the glass spherical substrate,
wherein the fibroblasts attach to the glass spherical substrate after culturing for a sufficient time and the mesenchymal stem cells do not attach to the glass spherical substrate.

### Claim 2 (depends on 1)

The method of claim 1, wherein the glass spherical substrate consists of an array of glass spherical substrates.

### Claim 3 (depends on 1)

The method of claim 1, wherein the shape of the glass spherical substrate is selected from the group consisting of a convex, a concave, and combinations thereof.

### Claim 4 (depends on 1)

The method of claim 1, wherein the glass spherical substrate is a ball.

### Claim 5 (depends on 1)

The method of claim 1, wherein the glass spherical substrate has a diameter of 58-300 μm.

### Claim 6 (depends on 1)

The method of claim 1, wherein the mixed population of cell types comprises human mesenchymal stem cells and fibroblasts.