# US 8,153,421 B2 · Prostate stem cell

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- Number: US 8,153,421 B2
- Document ID: US8153421B2
- Status: Granted
- Kind: B2
- Application number: 10/593,128
- Filed: 2005-03-18
- Published: 2008-09-25
- Granted: 2012-04-10
- Earliest claimed: 2004-03-19
- Assignees: Procure Therapeutics Limited
- Inventors: Norman James Maitland; Anne Collins
- Web page: https://patentgrep.com/patent/US8153421B2

## CPC

- G01N 33/5011: {for testing antineoplastic activity}
- A61P 13/08: of the prostate
- A61P 35/00: Antineoplastic agents
- C12N 5/0695: {Stem cells; Progenitor cells; Precursor cells}
- G01N 33/57555: of the prostate
- A61K 2039/5152: {Tumor cells}
- C12N 2533/90: Substrates of biological origin, e.g. extracellular matrix, decellularised tissue
- 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)
- 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)
- A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D)

## Abstract

We describe a method for the isolation of prostate stem cells, typically prostate stem cells which express CD 133 antigen; stem cells and cancer stem cells isolated by the method and their use.

## Claims

### Claim 1

A method for the selective enrichment of prostate cancer stem cells which express CD133, CD44, and high levels of α2β1 integrin which comprises the following steps:
i) providing a cell preparation comprising prostate cancer stem cells derived from prostate tissue;
ii) providing cell culture conditions which allow the maintenance of said prostate cancer stem cells in culture and the binding of said prostate cancer stem cells to a collagen based matrix;
iii) selecting said bound cells for expression of CD133 and isolating bound cells that express CD133 antigen, CD44 antigen, and α2β1 integrin.

### Claim 2 (depends on 1)

A method according to claim 1 wherein said method includes the additional steps of:
iv) culturing prostate cancer stem cells which express CD133 antigen in culture medium comprising granulocyte macrophage colony stimulating factor (GM-CSF), stem cell factor (SCF) and leukaemia inhibitory factor (LIF); and
v) passaging the prostate cancer stem cells in (i) in a serum free medium.

### Claim 3 (depends on 1)

A method according to claim 1 wherein said selected cells express human epithelial antigen.

### Claim 4 (depends on 1)

A method according to claim 1 wherein said prostate cancer stem cells are metastatic.

### Claim 5 (depends on 1)

A method according to claim 1 wherein said prostate cancer stem cells are from a primary prostate tumour.

### Claim 6 (depends on 1)

A method according to claim 1 wherein said collagen based matrix comprises collagen I.

### Claim 7 (depends on 1)

A prostate cancer stem cell obtainable by the method of claim 1, wherein said prostate cancer stem cells have high in vitro proliferative potential, have higher colony forming efficiency than α2β1 integrinlow CD133− prostate cells and can form cancerous prostatic-like acini in an immune-compromised non-human animal model.

### Claim 8 (depends on 7)

A prostate cancer stem cell according to claim 7 wherein said stem cell is cloned.

### Claim 9

A cell culture of prostate cancer stem cells wherein said cells express CD133 antigen, CD44 antigen, and α2β1 integrin wherein said prostate cancer stem cells have high in vitro proliferative potential, have higher colony forming efficiency than α2β1 integrinlow CD 133− prostate cells and can form cancerous prostatic-like acini in an immune-compromised non-human animal model.

### Claim 10 (depends on 9)

A cell culture according to claim 9 wherein said prostate cancer stem cells express human epithelial antigen.