# US 9,750,826 B2 · Method for preparing patient-specific glioblastoma animal model, and uses thereof

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- Number: US 9,750,826 B2
- Document ID: US9750826B2
- Status: Granted
- Kind: B2
- Application number: 14/409,668
- Filed: 2013-06-21
- Published: 2015-04-16
- Granted: 2017-09-05
- Earliest claimed: 2012-06-21
- Assignees: Samsung Life Public Welfare Foundation
- Applicants: Samsung Life Public Welfare Foundation
- Inventors: Do Hyun Nam; Ju Yuon Jin; Kyeung Min Joo
- Web page: https://patentgrep.com/patent/US9750826B2

## CPC

- A61K 49/0008: {Screening agents using (non-human) animal models or transgenic animal models or chimeric hosts, e.g. Alzheimer disease animal model, transgenic model for heart failure}
- A01K 67/0271: Chimeric vertebrates, e.g. comprising exogenous cells
- A01K 2227/105: Murine
- A01K 2267/0331: Animal model for proliferative diseases
- A01K 2267/0356: Animal model for processes and diseases of the central nervous system, e.g. stress, learning, schizophrenia, pain, epilepsy
- 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)
- A01K: ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS

## Abstract

Provided is a method for producing a glioblastoma mouse model and the mouse model produced thereby, the method including the steps of: (a) dividing a glioblastoma tissue, isolated from a patient, into 4 or more sections, and collecting one or more pieces from each of the sections; (b) dissociating a mixture of the collected pieces into glioblastoma cells as single cells; and (c) orthotopically transplanting a graft sample containing the glioblastoma cells obtained in step (b), into the brain of an immunodeficient mouse. Further provided are a method of screening a glioblastoma therapeutic agent using the mouse model and a method of providing information for selection of a patient-specific glioblastoma therapy using the mouse model. The glioblastoma mouse model shows the same genetic, morphological and pathological characteristics as those of the parental tumor, and it allows screening patient-specific glioblastoma therapeutic agent or selecting safer and more effective patient-specific glioblastoma therapy.

## Claims

### Claim 1

A method for producing a glioblastoma mouse model comprising the steps of:
(a) dividing a whole glioblastoma tissue, isolated from a patient, into 4 or more sections, collecting one or more pieces from each of the 4 or more sections, and homogenously mixing the collected pieces to prepare a homogenous mixture of the glioblastoma tissue pieces collected from each of the 4 or more sections of the whole glioblastoma tissue;
(b) dissociating single cells of glioblastoma including glioblastoma stem cells from the homogenously mixed glioblastoma tissue pieces by sequentially performing mechanical dissociation and enzymatic dissociation with an enzyme mixture including protease and DNase;
(b)′ removing erythrocytes by Percoll gradient centrifugation from a graft sample, which contains the glioblastoma cells as single cells obtained in step (b); and
(c) orthotopically transplanting a graft sample comprising 1×105 to 2.0×105 glioblastoma cells obtained in step (b) in a volume of 5-10 μl, into the brain of an immunodeficient mouse by injecting the sample into the brain at the location of 1.5-2.5 mm left and 0.5-1.5 mm anterior to the bregma and 1.8-2.2 mm deep from the dura.

### Claim 2 (depends on 1)

The method of claim 1, wherein each of the one or more pieces has a volume of 1-8 mm3 and is collected from the 4 or more sections of the glioblastoma tissue in step (a).

### Claim 3 (depends on 1)

The method of claim 1, wherein the volume of the mixture of the pieces collected in step (b) is 30-100% of the volume of the glioblastoma tissue isolated from the patient.

### Claim 4 (depends on 1)

The method of claim 1, wherein the immunodeficient mouse is NOG (NOD/Shi-scid, IL-2Ry KO) mouse.

### Claim 5 (depends on 1)

The method of claim 1, wherein the glioblastoma mouse model shows the same genetic and morphological characteristics as those of glioblastoma of the patient.