# US 12,180,442 B2 · Method for aggregating cell mass and device for aggregating cell mass

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- Number: US 12,180,442 B2
- Document ID: US12180442B2
- Status: Granted
- Kind: B2
- Application number: 16/765,495
- Filed: 2018-11-30
- Published: 2020-10-08
- Granted: 2024-12-31
- Earliest claimed: 2017-11-30
- Assignees: Public University Corporation Yokohama City University; Japan Aerospace Exploration Agency
- Applicants: Public University Corporation Yokohama City University; Japan Aerospace Exploration Agency
- Inventors: Hideki Taniguchi; Tomomi Tadokoro; Tetsuya Sakashita; Satoshi Matsumoto; Satoshi Adachi; Akira Higashibata
- Web page: https://patentgrep.com/patent/US12180442B2

## CPC

- C12M 21/08: {for producing artificial tissue or for ex-vivo cultivation of tissue (prostheses A61F2/00, grafts A61L27/00)}
- C12M 35/04: {Mechanical means, e.g. sonic waves, stretching forces, pressure or shear stimuli}
- C12M 27/10: {Rotating vessel}
- C12M 37/00: {Means for sterilizing, maintaining sterile conditions or avoiding chemical or biological contamination (C12M23/38 takes precedence; filtration in general and filters per se B01D24/00-B01D41/00; autoclaves B01J3/04; treatment of microorganisms with electrical or wave energy C12N13/00)}
- C12M 41/40: {of pressure}
- C12M 41/48: {Automatic or computerized control (automatic analysis G01N35/00)}
- C12M 47/02: {Separating microorganisms from the culture medium; Concentration of biomass (separating microorganisms from their culture media C12N1/02)}
- C12N 1/00: Microorganisms; Compositions thereof (medicinal preparations containing material from protozoa, bacteria or viruses A61K35/66, from algae A61K36/02, from fungi A61K36/06; preparing medicinal bacterial antigen or antibody compositions, e.g. bacterial vaccines, A61K39/00); Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N 5/0062: {General methods for three-dimensional culture}
- C12N 5/0697: {Artificial constructs associating cells of different lineages, e.g. tissue equivalents (blood vessels C12N5/0691)}
- C12N 2535/10: Patterned coating
- C12M: APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; {APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS}
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)

## Abstract

A method for aggregating cell masses includes performing a cell mass aggregating step of rotating a rotating body containing a specific gravity adjustment solution and cell masses to aggregate the cell masses, the specific gravity adjustment solution having biocompatibility, the cell masses having a lower specific gravity than the specific gravity adjustment solution.

## Claims

### Claim 1

A method for aggregating cell masses, comprising:
the step of attaching a first end and a second end of a tubular shaped base material that is an artificial blood vessel or a regeneration blood vessel, respectively, to a first mounting portion and a second mounting portion that are arranged on a central axis of the rotating body and are spaced apart from each other, to linearly extend the base material on the central axis from the first end to the second end without bending the base material and to connect the first mounting portion and the second mounting portion via the artificial base material; and
a cell mass aggregating step of rotating at the rotation speed a rotating body containing a specific gravity adjustment solution and cell masses and aggregating the cell masses on an outer surface of the base material by centripetal force acting on the cell masses due to rotation of the rotating body and difference in specific gravity between the specific gravity adjustment solution and the cell masses, the specific gravity adjustment solution having biocompatibility, the cell masses having a lower specific gravity than the specific gravity adjustment solution, and the rotation speed being selected to cause cell aggregation by the centripetal force.

### Claim 2 (depends on 1)

The method for aggregating cell masses according to claim 1, wherein in the cell mass aggregating step, the rotating body is rotated about the central axis of the rotating body to aggregate the cell masses around the central axis.

### Claim 3 (depends on 1)

The method for aggregating cell masses according to claim 1, wherein the cell mass aggregating step is performed while supplying a liquid culture medium into the base material.

### Claim 4 (depends on 1)

The method for aggregating cell masses according to claim 1, further comprising a deaeration step of rotating the rotating body in which air is contained, collecting the air on the base material by centripetal force acting on the air due to rotation of the rotating body and difference in specific gravity between the specific gravity adjustment solution and the air, and removing the air from the rotating body by the air passing through the base material having gas permeability.

### Claim 5 (depends on 4)

The method for aggregating cell masses according to claim 4, wherein the deaeration step is performed before the cell masses are injected into the rotating body.

### Claim 6 (depends on 1)

The method for aggregating cell masses according to claim 1, wherein the specific gravity adjustment solution includes a liquid culture medium.

### Claim 7 (depends on 1)

The method for aggregating cell masses according to claim 1, the method comprising completely covering the outer surface of the base material with uniformly arranged vascular organoids using the centripetal force, and culturing vascular organoids to form and mature a surface vascular network.