# US 9,752,128 B2 · Methods of generating cells exhibiting phenotypic plasticity

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- Number: US 9,752,128 B2
- Document ID: US9752128B2
- Status: Granted
- Kind: B2
- Application number: 13/377,296
- Filed: 2010-06-10
- Published: 2012-04-12
- Granted: 2017-09-05
- Earliest claimed: 2009-06-10
- Assignees: Stephen Sanig Research Institute Ltd.
- Applicants: Galina Kaseko; Tohsak L. Mahaworasilpa
- Inventors: Galina Kaseko; Tohsak L. Mahaworasilpa
- Web page: https://patentgrep.com/patent/US9752128B2

## CPC

- C12N 5/16: Animal cells
- C12N 5/0634: {Cells from the blood or the immune system}
- C12N 5/0635: {B lymphocytes}
- C12N 5/0636: {T lymphocytes}
- C12N 5/0646: {Natural killers cells [NK], NKT cells}
- 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 relates to cellular differentiation and, in particular to hybrid cells that exhibit phenotypic plasticity and methods for producing these cells. The invention also relates to methods for generating specific cells of a desired phenotype. The invention still further relates to methods of producing hybrid cells with a capacity to de-differentiate into an earlier progenitor state. The invention further contemplates the use of hybrid cells in a range of applications, for example tissue generation.

## Claims

### Claim 1

A method of generating CD8+ T-cell from a tri-hybrid cell, said method comprising the steps of
(i) hybridizing a first, a second and a third cell to produce a tri-hybrid cell that exhibits genetic stability, phenotypic plasticity and cellular markers of said first, second and third cell, wherein a myeloma cell is not included in the hybridization, wherein the first cell is an immortalized myeloid cell, wherein the second cell is selected from the group consisting of a CD19+ mature B cells, a CD20−CD72+ early B cells, a CD20+CD72+ activated B cells and a CD19−CD5+ antigen experienced B cells, and wherein the third cell is selected from group consisting of a CD4+ helper T cell, a CD3+CD5+antigen-experienced T cells, CD3+ T cell and a CD4+CD8+ T cells;
(ii) determining cellular markers of said first, second and third cell in the tri-hybrid cell to establish the presence in the genetically stable tri-hybrid cell of all the cellular markers of said first, second and third cell; and
(iii) culturing said genetically stable tri-hybrid cell with human thymic stromal cells to produce a CD8 expressing T-cell.

### Claim 2 (depends on 1)

The method according to claim 1, wherein said first cell is a K562 cell.

### Claim 3 (depends on 1)

The method according to claim 1, wherein said second cell is a WIL2NS cell and said third cell is a MOLT4cell.

### Claim 4 (depends on 1)

The method according to claim 1, wherein said second cell is a WIL2NS cell.

### Claim 5 (depends on 1)

The method according to claim 1, wherein said third cell is a MOLT4cell.

### Claim 6 (depends on 1)

The method according to claim 1, wherein said first cell is a K562cell, said second cell is a WIL2NS cell and said third cell is a MOLT4 cell.

### Claim 7 (depends on 1)

The method according to claim 1, wherein said first cell is a K562cell, said second cell is a primary B cell and said third cell is a primary T cell.