# US 11,441,123 B2 · Method for evaluating quality of human mesenchymal stem cell, and monoclonal antibody for use in said method

> Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt

- Number: US 11,441,123 B2
- Document ID: US11441123B2
- Status: Granted
- Kind: B2
- Application number: 17/190,192
- Filed: 2021-03-02
- Published: 2021-07-29
- Granted: 2022-09-13
- Earliest claimed: 2014-08-01
- Assignees: Purec Co., Ltd.
- Applicants: Purec Co., Ltd.
- Inventors: Yumi Iyoku; Hideyuki Okano; Yo Mabuchi
- Web page: https://patentgrep.com/patent/US11441123B2

## CPC

- C12N 5/0663: {Bone marrow mesenchymal stem cells (BM-MSC)}
- C07K 16/28: against receptors, cell surface antigens or cell surface determinants
- C12N 15/02: Preparation of hybrid cells by fusion of two or more cells, e.g. protoplast fusion {(monoclonal antibodies C07K16/00; apparatus for cell fusion C12M)}
- C12P 1/00: Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes
- C12Q 1/04: Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor {(C12Q1/6897 takes precedence)}
- C12Q 1/6881: for tissue or cell typing, e.g. human leukocyte antigen [HLA] probes
- C12Y 207/10001: Receptor protein-tyrosine kinase (2.7.10.1)
- G01N 33/56966: {Animal cells}
- G01N 2333/705: Assays involving receptors, cell surface antigens or cell surface determinants
- G01N 2333/912: transferring phosphorus containing groups, e.g. kinases (2.7)
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00)
- C12P: FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE {(brewing of beer C12C; producing vinegar C12J; producing specific peptides or proteins C07K; producing enzymes C12N9/00; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification C12N15/00; measuring or testing processes involving enzymes or microorganisms C12Q; measuring or testing processes involving nucleic acid amplification reactions C12Q1/6844; fermentation processes to form a food composition, A21 or A23; compounds in general, see the relevant compound class, e.g. C01, C07)}
- C12Q: MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS (immunoassay G01N33/53); COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Y: ENZYMES
- 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)

## Abstract

The present invention aims to obtain a method for quality evaluation of human mesenchymal stem cells, a method for isolation, selection and culture of human mesenchymal stem cells, a cell population of rapidly proliferating human mesenchymal stem cells, as well as monoclonal antibodies that specifically recognize rapidly proliferating human mesenchymal stem cells. From a cell population containing human mesenchymal stem cells, rapidly proliferating human mesenchymal stem cells are isolated, selected and cultured. The abundance ratio of cells expressing Ror2 or Fzd5 in the cell population thus isolated, selected and cultured is quantified to determine whether or not each cell population is acceptable.

## Claims

### Claim 1

A method for selecting and isolating proliferating human mesenchmal stem cells, which comprises:
a step 1 where from a cell population containing primary human tissue-derived mesenchymal stem cells, rapidly proliferating human mesenchymal stem cells which are Ror231 cells are single-cell cultured on a culture vessel surface to produce a cell population from said single-cell; and
a step 2 where the abundance ratio of cells expressing Ror2 collected from the cell population obtained in the step 1 is quantified and
a step 3 selecting and isolating only the cell population to be at least 63% Ror2−.

### Claim 2 (depends on 1)

The method according to claim 1, wherein the cells expressing Ror2 are quantified by immunostaining.

### Claim 3 (depends on 1)

The method according to claim 1, wherein the cells expressing Ror2 are quantified by using anti-Ror2 monoclonal antibody.

### Claim 4 (depends on 1)

The method according to claim 1, wherein the cells expressing mRNA of Ror2 are quantified by quantitative PCR.

### Claim 5 (depends on 1)

The method according to claim 1, wherein the step 1 comprises sorting Ror2− cells by analyzing cells stained with anti-Ror2 monoclonal antibody by FCM.

### Claim 6 (depends on 5)

The method according to claim 5, wherein the sorting comprises seeding the Ror2− cells in wells of a multi-well culture plate and then isolating and selecting the cells in each well reaching confluence upon culture.

### Claim 7 (depends on 1)

The method of claim 1, wherein said percentage is at least 65%.