# US 11,898,162 B2 · Reprogramming somatic cells into pluripotent cells using a vector encoding Oct4 and Sox2

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- Number: US 11,898,162 B2
- Document ID: US11898162B2
- Status: Granted
- Kind: B2
- Application number: 16/818,958
- Filed: 2020-03-13
- Published: 2020-07-30
- Granted: 2024-02-13
- Earliest claimed: 2007-03-23
- Assignees: Wisconsin Alumni Research Foundation
- Applicants: Wisconsin Alumni Research Foundation
- Inventors: James A. Thomson; Junying Yu
- Web page: https://patentgrep.com/patent/US11898162B2

## CPC

- A61P 3/10: for hyperglycaemia, e.g. antidiabetics
- C12N 5/0607: {Non-embryonic pluripotent stem cells, e.g. MASC (induced pluripotent stem cells C12N5/0696)}
- A61P 25/00: Drugs for disorders of the nervous system
- A61P 25/16: Anti-Parkinson drugs
- C12N 5/0696: {Artificially induced pluripotent stem cells, e.g. iPS}
- C12N 5/0606: {Pluripotent embryonic cells, e.g. embryonic stem cells [ES] (embryonic germ cells C12N5/0611, induced pluripotent stem cells C12N5/0696)}
- C12N 2501/60: Transcription factors
- C12N 2501/602: Sox-2
- C12N 2501/603: Oct-3/4
- C12N 2501/604: Klf-4
- C12N 2501/605: Nanog
- C12N 2501/606: c-Myc
- C12N 2501/608: Lin28
- C12N 2502/99: genetically modified cells
- C12N 2510/00: Genetically modified cells
- C12N 2799/027: where the vector is derived from a retrovirus
- A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- 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 methods for reprogramming a somatic cell to pluripotency by administering into the somatic cell at least one or a plurality of potency-determining factors. The invention also relates to pluripotent cell populations obtained using a reprogramming method.

## Claims

### Claim 1

A method of reprogramming primate somatic cells, the method comprising the steps of:
introducing a retroviral or non-integrating vector comprising a polynucleotide encoding potency-determining factors into the primate somatic cells under conditions sufficient to express the potency-determining factors, thereby reprogramming the cells, wherein the potency-determining factors comprise Oct-4 and Sox2, and do not comprise c-Myc and Klf4; and
culturing the primate somatic cells under embryonic stem cell culture conditions to obtain pluripotent reprogrammed cells.

### Claim 2 (depends on 1)

The method of claim 1, wherein the primate somatic cells are obtained from a postnatal individual.

### Claim 3 (depends on 1)

The method of claim 1, wherein the primate somatic cells are obtained by in vitro differentiation of a stem cell.

### Claim 4 (depends on 1)

The method of claim 1, wherein the potency-determining factors are introduced to the somatic cells as a reprogramming sequence in which a nucleic acid sequence encoding the potency-determining factor is operably linked to a heterologous promoter.

### Claim 5 (depends on 1)

The method of claim 1, wherein the potency-determining factors are Oct-4, Sox2, and at least one of Nanog and Lin28.

### Claim 6 (depends on 1)

The method of claim 1, wherein the potency-determining factors are Oct-4 and Sox2.

### Claim 7 (depends on 1)

The method of claim 1, wherein the reprogrammed cells (i) express a cell marker selected from the group consisting of Oct-4, SSEA3, SSEA4, Tra-1-60 and Tra-1-81; (ii) exhibit morphology characteristic of pluripotent cells; and (iii) form teratomas when introduced into an immunocompromised animal.

### Claim 8

A method of reprogramming primate somatic cells, the method comprising the steps of:
introducing a retroviral or non-integrating vector comprising a polynucleotide encoding potency-determining factors into the primate somatic cells under conditions sufficient to express the potency-determining factors, thereby reprogramming the cells, wherein the potency-determining factors comprise Oct-4, Sox2, Nanog, and Lin28; and
culturing the primate somatic cells under embryonic stem cell culture conditions to obtain reprogrammed cells having a higher potency level than the primate somatic cells.