# US 2018/0023055 A1 · Method for culturing primary cells

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- Number: US 2018/0023055 A1
- Document ID: US20180023055A1
- Status: Published application
- Kind: A1
- Application number: 15/444,588
- Filed: 2017-02-28
- Published: 2018-01-25
- Earliest claimed: 2016-07-22
- Assignees: Osaka Prefectural Hospital Organization
- Applicants: Osaka Prefectural Hospital Organization
- Inventors: Norikatsu MIYOSHI; Shiki FUJINO; Masayuki OHUE; Masayoshi YASUI
- Web page: https://patentgrep.com/patent/US20180023055A1

## CPC

- C12N 5/0693: {Tumour cells; Cancer cells}
- C12N 5/0679: {Cells of the gastro-intestinal tract}
- G01N 33/5005: {involving human or animal cells (immunoassay G01N33/56966; immunoassays of protozoa G01N33/56905; protozoa in screening assays C12Q1/025)}
- G01N 33/5011: {for testing antineoplastic activity}
- C12N 2503/00: Use of cells in diagnostics
- C12N 2503/02: Drug screening
- C12N 2509/10: Mechanical dissociation
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- 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)

## Abstract

Method for producing a primary culture of cancer cells, comprising: (a) fragmenting cancer tissues derived from a living body and removing impurities from the fragmented cancer tissues, (b) subjecting the tissue masses obtained in (a) to suspension culture, and (c) subjecting the culture obtained in (b) to adherent culture.

## Claims

### Claim 1

A method for producing a primary culture of cancer cells, comprising:
(a) fragmenting cancer tissues derived from a living body and removing impurities from the fragmented cancer tissues;
(b) subjecting the tissue masses obtained in (a) to suspension culture; and
(c) subjecting the culture obtained in (b) to adherent culture.

### Claim 2 (depends on 1)

The method according to claim 1, wherein
the removing of the impurities is performed by sieving, and.
a lower limit size and an upper limit size of cell masses obtained by the sieving are 20 μm to 100 μm and 200 μm to 500 μm, respectively.

### Claim 3 (depends on 1)

The method according to claim 1, wherein the suspension culture is performed for 24 to 48 hours.

### Claim 4 (depends on 1)

The method according to claim 1, further comprising:
subjecting the culture obtained in (c) to subculture.

### Claim 5 (depends on 1)

A method for elucidating a molecular mechanism of cancer, performing a drug-susceptibility test, or screening a pharmaceutical, comprising
using the primary culture of cancer cells obtained the method according to claim 1.

### Claim 6 (depends on 1)

A kit for elucidating a molecular mechanism of cancer, performing a drug-susceptibility test, or screening a pharmaceutical, comprising
the primary culture of cancer cells obtained by the method. according to claim 1.