# US 10,501,726 B2 · Population of isolated chicken cells reprogrammed from somatic cells

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- Number: US 10,501,726 B2
- Document ID: US10501726B2
- Status: Granted
- Kind: B2
- Application number: 15/164,618
- Filed: 2016-05-25
- Published: 2016-11-17
- Granted: 2019-12-10
- Earliest claimed: 2011-06-10
- Assignees: University of Georgia Research Foundation, Inc.
- Applicants: University of Georgia Research Foundation, Inc.
- Inventors: Steven L Stice; Franklin West; Yangqing Lu
- Web page: https://patentgrep.com/patent/US10501726B2

## CPC

- C12N 5/0696: {Artificially induced pluripotent stem cells, e.g. iPS}
- A01K 67/0271: Chimeric vertebrates, e.g. comprising exogenous cells
- A01K 67/0275: Genetically modified vertebrates, e.g. transgenic
- A01K 2207/12: Animals modified by administration of exogenous cells
- A01K 2227/30: Bird
- A01K 2267/02: Animal zootechnically ameliorated
- C12N 2501/60: Transcription factors
- C12N 2501/602: Sox-2
- C12N 2501/603: Oct-3/4
- C12N 2501/605: Nanog
- C12N 2501/608: Lin28
- C12N 2502/99: genetically modified cells
- C12N 2506/02: from embryonic cells
- C12N 2506/1307: from adult fibroblasts
- C12N 2760/16111: Influenzavirus A, i.e. influenza A virus
- C12N 2760/18111: Avulavirus, e.g. Newcastle disease virus
- C12N 2999/007: Technological advancements, e.g. new system for producing known virus, cre-lox system for production of transgenic animals
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- A01K: ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS

## Abstract

The present invention relates to the production of avian induced pluripotent stem cells from non-pluripotent somatic cells, including embryonic fibroblasts and adult somatic cells. In this method, avian (including quail or chicken) somatic cells are reprogrammed into a state closely resembling embryonic stem cells including the expression of key stem cell markers alkaline phosphatase, etc. by transfecting/transducing the non-stem cells with genes (preferably using a non-integrating vector as otherwise described herein or alternatively an integrating vector, such a lentiviral vector, retroviral vector or inducible lentiviral vector, among others) which express at least nanog, Lin28 and cMyc. In preferred aspects of the invention, the transfected/transduced vectors express nanog, Lig28, cMyc, Oct 4 (POU5F1 or PouV), SOX2 and KLF4. The induced stem cells which are produced contribute to all 3 germ layers, the trophectoderm and in certain aspects, the gonad in chimeric offspring.

## Claims

### Claim 1

A population of isolated chicken cells which have been reprogrammed from chicken somatic cells, wherein said isolated chicken cells comprise exogenous nucleic acid sequences encoding OCT4, SOX2, LIN28 and NANOG genes, and wherein said isolated cells exhibit the following characteristics:
the cells express Alkaline Phosphotase, SSEA1, EMA1, Chicken Vasa Homolog (CVH), DAZL, CXCR4, C-KIT and telomerase; and
the cells are capable of differentiating into all 3 germ layers in vitro.

### Claim 2 (depends on 1)

The population of isolated cells according to claim 1 wherein said chicken somatic cells are embryonic chicken somatic cells.

### Claim 3 (depends on 1)

The population of isolated cells according to claim 1 wherein said chicken somatic cells are embryonic chicken fibroblast cells.

### Claim 4 (depends on 1)

The population of isolated cells according to claim 1 wherein said chicken somatic cells are adult chicken somatic cells.

### Claim 5 (depends on 1)

The population of isolated cells according to claim 1 wherein said chicken somatic cells are adult chicken fibroblast cells.

### Claim 6 (depends on 1)

The population of isolated cells according to claim 1 wherein said chicken somatic cells are reprogrammed by transfecting or transducing said somatic cells with at least one vector comprising said exogenous genes.

### Claim 7 (depends on 1)

The population of isolated cells according to claim 1 wherein said exogenous genes are avian or mammalian genes each of which has a sequence homology of at least about 50% of the identical gene of a chicken.

### Claim 8 (depends on 1)

The population according to claim 1 wherein said exogenous genes are chicken, mouse, human, pig or cow genes.

### Claim 9 (depends on 1)

The population according to claim 1 wherein said chicken somatic cells are transfected or transduced with vectors wherein each of said vectors comprises a single exogenous gene.

### Claim 10 (depends on 1)

The population of cells according to claim 1 wherein said exogenous genes include C-MYC.