WO2017033958A1 - Cell ink, cell ink production container, cell ink production kit, cell ink production method, ink cartridge, sterically formed object production method, and sterically formed object - Google Patents

Cell ink, cell ink production container, cell ink production kit, cell ink production method, ink cartridge, sterically formed object production method, and sterically formed object Download PDF

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Publication number
WO2017033958A1
WO2017033958A1 PCT/JP2016/074608 JP2016074608W WO2017033958A1 WO 2017033958 A1 WO2017033958 A1 WO 2017033958A1 JP 2016074608 W JP2016074608 W JP 2016074608W WO 2017033958 A1 WO2017033958 A1 WO 2017033958A1
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Prior art keywords
cell
ink
container
dimensional
cell ink
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PCT/JP2016/074608
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French (fr)
Japanese (ja)
Inventor
巌 和賀
洋子 江尻
Original Assignee
Ｎｅｃソリューションイノベータ株式会社
株式会社クラレ
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Application filed by Ｎｅｃソリューションイノベータ株式会社, 株式会社クラレ filed Critical Ｎｅｃソリューションイノベータ株式会社
Priority to JP2017536456A priority Critical patent/JP6629332B2/en
Publication of WO2017033958A1 publication Critical patent/WO2017033958A1/en

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • 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
    • C12M33/00—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • 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
    • C12M25/00—Means for supporting, enclosing or fixing the microorganisms, e.g. immunocoatings
    • C12M25/02—Membranes; Filters
    • C12M25/04—Membranes; Filters in combination with well or multiwell plates, i.e. culture inserts

Definitions

  • the present invention relates to a cell ink, a cell ink production container, a cell ink production kit, a cell ink production method, an ink cartridge, a method for producing a three-dimensional object, and a three-dimensional object.
  • Patent Document 1 An attempt has been made to produce organs and the like by three-dimensionally culturing pluripotent cells such as iPS cells and ES cells.
  • the present invention relates to a cell ink capable of modeling a three-dimensional structure, a cell ink manufacturing container capable of manufacturing a cell ink, a cell ink manufacturing kit, a method of manufacturing a cell ink, an ink cartridge, and a method of manufacturing a three-dimensional structure. And a three-dimensionally shaped object.
  • the cell ink of the present invention is characterized by containing a cell mass.
  • the cell ink production container of the present invention includes a bottomed cylindrical container, The bottom of the container includes one or more openings;
  • the opening is a culture chamber for culturing a cell mass,
  • the size of the diameter of the opening on the outer surface side of the bottom is not more than the size of the diameter of the ink discharge port of the three-dimensional modeling apparatus that performs three-dimensional modeling by discharging ink.
  • the cell ink production kit of the present invention includes the cell ink production container of the present invention and a collection container for collecting the cell mass,
  • the collection container has a bottomed cylindrical shape, and can accommodate the cell ink production container therein.
  • the method for producing a cell ink of the present invention includes a culture step of culturing cells using the cell ink production container of the present invention, and a recovery step of collecting the cultured cell mass,
  • a culture step of culturing cells using the cell ink production container of the present invention and a recovery step of collecting the cultured cell mass
  • In the culturing step cells are cultured in the opening of the cell ink production container in the presence of a culture solution
  • the collecting step the cell mass led out of the cell ink production container is collected from the opening.
  • the cell ink of the present invention is obtained by the method for producing the cell ink of the present invention.
  • An ink cartridge for a three-dimensional modeling apparatus that performs three-dimensional modeling by discharging the ink of the present invention includes the cell ink of the present invention.
  • the method for producing a three-dimensional structure of the present invention includes a three-dimensional modeling process using cell ink,
  • the three-dimensional modeling step is based on the three-dimensional data of the modeling object, and discharges the cell ink from the ink discharge port of the three-dimensional modeling apparatus that performs ink three-dimensional modeling by discharging ink, and models the three-dimensional modeling object,
  • the cell ink is at least one of the cell ink of the present invention and the cell ink obtained by the method for producing the cell ink of the present invention.
  • the three-dimensional structure of the present invention is obtained by the method for manufacturing a three-dimensional structure of the present invention.
  • FIG. 1 (A) is a schematic perspective view showing an example of the cell ink production container of the present invention
  • FIG. 1 (B) is a schematic cross-sectional view as seen from the II direction in FIG. 1 (A). is there.
  • FIG. 2 (A) is a schematic exploded perspective view showing an example of the cell ink production kit of the present invention
  • FIG. 2 (B) shows a state in which the production container is housed in the collection container in the production kit.
  • FIG. 2C is a schematic perspective view
  • FIG. 2C is a schematic cross-sectional view as seen from the II-II direction in FIG. 2B.
  • downward direction means the bottom surface direction of the bottomed cylindrical container
  • upward direction means the direction opposite to the bottom surface direction of the bottomed cylindrical container, that is, the opening direction.
  • the vertical length is also called height.
  • inside and inside means a region surrounded by a bottom surface and a cylindrical side wall of the bottomed cylindrical container
  • outside means a bottom surface and a cylinder of the bottomed cylindrical container. Means a region not surrounded by a side wall.
  • the cell ink of the present invention includes a cell mass.
  • the cell ink of the present invention is characterized by containing the cell mass, and other configurations and conditions are not particularly limited. Since the cell ink of the present invention includes the cell mass, for example, when modeling a three-dimensional model having the same size, the cell ink can be modeled in a shorter time than a cell ink including a single cell. In addition, according to the cell ink of the present invention, for example, since a three-dimensional model can be modeled in a shorter time, cell death of cells constituting the modeled three-dimensional model can be suppressed.
  • the density of cell adhesion molecules on the cell surface of the cells constituting the cell mass is not impaired.
  • a food with high freshness and close to the original taste can be modeled.
  • an organ in which the organ function is maintained can be modeled.
  • the cell ink of the present invention for example, when a three-dimensional object is modeled by a three-dimensional modeling apparatus (hereinafter, also referred to as “inkjet 3D printer”) that performs three-dimensional modeling by discharging ink described later, It is possible to reduce clogging of the ink discharge port of the ink jet 3D printer.
  • a three-dimensional modeling apparatus hereinafter, also referred to as “inkjet 3D printer”
  • the type of cells constituting the cell mass is not particularly limited, and can be appropriately determined according to, for example, the type of three-dimensional modeled object to be modeled using the cell ink.
  • Examples of the cell include the same cell as the target cell described later or a different cell.
  • examples of the cells include parenchymal cells such as hepatocytes, sinusoidal wall cells, myofibroblasts, bile duct epithelial cells, and connective tissue cells.
  • the cell mass means an aggregate formed from a plurality of the cells, and can also be referred to as, for example, a spheroid.
  • the cell mass may be composed of, for example, one type of cell or may be composed of two or more types of cells.
  • the origin of the cell is not particularly limited, and examples thereof include humans, non-human mammals, birds, reptiles, and seafood.
  • the number of cells constituting the cell mass is not particularly limited, and is, for example, 10 to 10,000, and is preferably 10 to 5000 because cell death of cells inside the cell mass can be suppressed.
  • the size of the cell mass is not particularly limited, and for example, the diameter is 20 to 900 ⁇ m, 50 to 500 ⁇ m.
  • the diameter of the cell mass may be, for example, a short diameter, a long diameter, or an average diameter, but is preferably a long diameter.
  • the diameter of the cell mass can more effectively suppress clogging of the ink discharge port of the ink jet 3D printer.
  • the diameter of the ink discharge port of the 3D printer (for example, the diameter, the same applies hereinafter) or less, more preferably 1/4 to 1 times the diameter of the ink discharge port of the ink jet 3D printer,
  • the length is in the range of 1/3 to 1 / 1.2 times.
  • the diameter of the ink discharge port may be, for example, a short diameter, a long diameter, or an average diameter, but is preferably a short diameter.
  • the diameter of the ink discharge port of the ink jet 3D printer is, for example, 20 to 4000 ⁇ m, 50 to 2000 ⁇ m.
  • the cell mass is preferably a uniform cell mass, and can further suppress clogging.
  • the diameter (D c ) of 90% or more, 95% or more, or 99% or more of the cell mass is ⁇ 30% (0.7% relative to the predetermined diameter (D p ).
  • the predetermined diameter can be appropriately set according to, for example, the diameter of the ink discharge port of the ink jet 3D printer.
  • the predetermined diameter is, for example, 20 to 4000 ⁇ m, 50 to 2000 ⁇ m. Since the diameter of the cell mass can further suppress clogging, in the cell mass contained in the cell ink, 85% or more, 90% or more, 95% or more, 96% or more, 97% or more, 98% or more, 99 % Or 100% of the cell mass preferably has a diameter of 95% or less, 90% or less, or 85% or less with respect to the diameter of the ink discharge port of the ink jet 3D printer, and 100% of cells It is more preferable that the lump has a diameter of 85% or less with respect to the diameter of the ink discharge port of the ink jet 3D printer.
  • the cell mass is, for example, a cell of a cell constituting the cell mass because adhesion between the cell masses is promoted after shaping the three-dimensional modeled object and cell death of the cells constituting the three-dimensional modeled product can be suppressed. It is preferred that the density of cell adhesion molecules on the surface is not impaired. Examples of the cell adhesion molecule include integrin, cadherin and the like. The density of the cell adhesion molecule may be, for example, the density of one type of cell adhesion molecule or the average density of two or more types of cell adhesion molecules.
  • the cell mass is, for example, after the modeling of the three-dimensional modeled object, the adhesion between the cell masses is promoted, and the cell death of the cells constituting the three-dimensional modeled product can be suppressed. Is preferably maintained.
  • the extracellular matrix include proteins such as collagen, proteoglycan, fibronectin and laminin, and sugars such as glycosaminoglycan.
  • the cell ink of the present invention may contain other components, for example.
  • the other components include a solvent for dispersing the cell mass, and specific examples include a cell ink solution described later.
  • the cell ink of the present invention can be produced by, for example, a cell ink production method described later.
  • the cell ink production container of the present invention includes a bottomed cylindrical container as described above, and the bottom of the container includes one or more openings,
  • the opening is a culture chamber for culturing a cell mass, and the size of the diameter of the opening on the outer surface side of the bottom is a diameter of an ink discharge port of a three-dimensional modeling apparatus that performs three-dimensional modeling by discharging ink.
  • the size is less than or equal to.
  • the diameter of the opening on the outer surface side of the bottom is equal to or smaller than the diameter of the ink discharge port of the three-dimensional modeling apparatus that performs three-dimensional modeling by discharging ink.
  • the other features and conditions are not particularly limited.
  • a cell ink capable of modeling a three-dimensional model can be manufactured as described later.
  • the diameter of the opening is equal to or less than the diameter of the ink discharge port of the three-dimensional modeling apparatus.
  • the size of the diameter of the cell mass derived from the opening to the outside of the manufacturing container is, for example, not more than the size of the diameter of the ink discharge port of the three-dimensional modeling apparatus, and is included in the cell ink.
  • the uniformity of the size of the cell mass is high. Therefore, when three-dimensional modeling is performed with the cell ink containing the cell mass, for example, clogging of the cell ink can be reduced at the ink discharge port of the three-dimensional modeling apparatus.
  • the cell ink manufacturing container of the present invention for example, it is possible to manufacture a cell ink in which clogging of ink discharge ports when a solid is modeled by the three-dimensional modeling apparatus is reduced.
  • the description of the cell ink of the present invention can be used for the production container of the present invention.
  • FIG. 1A is a schematic perspective view of the production container of the present embodiment
  • FIG. 1B is a schematic cross-sectional view as viewed from the II direction in FIG.
  • the manufacturing container 10 of the present embodiment includes a bottomed cylindrical container 1, and the container 1 includes a bottom portion 2, a side wall 4, and an edge 5.
  • the bottom 2 is formed at the lower end of the side wall 4, and the edge 5 is formed at the upper end of the side wall 4 so as to be integrated with the side wall 4.
  • the edge 5 protrudes outward of the side wall 4 and becomes, for example, a gripping part that grips the manufacturing container 10.
  • the bottom 2 of the container 1 includes a plurality of openings 3, and the openings 3 are tapered from the inner surface side to the outer surface side of the bottom portion 2.
  • the cell ink is produced, for example, by introducing a culture solution and cells into the inside of the bottomed cylindrical container 1 and culturing the cells and the cell mass formed from the cells at the opening 3. Can be implemented. For this reason, the opening 3 can also be called a culture chamber for culturing the cell mass. The cells and the cell mass will be described later.
  • the manufacturing container 10 of this embodiment contains the edge 5, the edge 5 is arbitrary structures and may or may not exist.
  • the material for forming the production container 10 is not particularly limited.
  • the manufacturing container 10 may be formed from, for example, one type of forming material, or may be formed from two or more types of forming materials. Moreover, in the latter case, each structure of the manufacturing container 10 may be comprised from a different forming material, for example.
  • the production container 10 includes one bottomed cylindrical container 1, but the present invention is not limited to this, and may include a plurality of bottomed cylindrical containers 1.
  • the plurality may be two or more, and the upper limit is not particularly limited. Examples of the plurality include 6, 12, 24, 48, 96, and 384.
  • the shape of the tube of the container 1 is not particularly limited, and examples thereof include a cylindrical shape and a rectangular tube shape.
  • the volume, inner diameter, height, and the like of the container 1 are not particularly limited.
  • the bottom 2 of the container 1 is planar, but the present invention is not limited to this, and may be curved, for example.
  • the bottom part 2 of the container 1 contains the some opening part 3, this invention is not limited to this, For example, you may include the one opening part 3.
  • FIG. When the bottom part 2 has the some opening part 3, the number of the opening parts 3 should just be two or more, and the upper limit in particular is not restrict
  • the number of openings 3 per unit area of the bottom 2 is not particularly limited, and is, for example, 10 to 1000 pieces / cm 2 and 20 to 1000 pieces / cm 2 .
  • the opening 3 is tapered from the inner surface side to the outer surface side of the bottom portion 2, but the present invention is not limited to this, and the opening portion 3 on the outer surface side of the bottom portion 2.
  • the diameter may be an arbitrary shape that is equal to or smaller than the diameter of the ink discharge port of the three-dimensional modeling apparatus that performs three-dimensional modeling by discharging ink.
  • the three-dimensional modeling apparatus that performs three-dimensional modeling by discharging the ink include an ink jet type three-dimensional modeling apparatus.
  • the ink discharge port include an opening of an ink jet head that discharges ink stored in an ink storage unit in the three-dimensional modeling apparatus.
  • the diameter (inner diameter) of the opening 3 is, for example, 50 to 1000 ⁇ m, 200 to 700 ⁇ m, or 300 to 500 ⁇ m.
  • a cell mass of about 200 ⁇ m can be produced.
  • the diameter of the opening 3 is not particularly limited, and can be appropriately set according to the size of the produced cell mass, for example.
  • the diameter of the opening 3 may be, for example, the same diameter from the inner side surface of the bottom part 2 toward the outer side surface or may be a different diameter, but is preferably the latter. In the latter case, the diameter of the opening 3 is preferably reduced from the inner surface to the outer surface of the bottom 2 because, for example, cells can be easily introduced into the opening 3.
  • the height in particular of the opening part 3 is not restrict
  • the height of the opening 3 is, for example, 1 to 10 times and 1.5 to 5 times the radius R of the cell mass having the desired diameter.
  • the radius R of the cell mass may be, for example, a short diameter, a long diameter, or an average diameter (hereinafter the same).
  • the edge 5 protrudes outward from the side wall 4, but the present invention is not limited to this, and can have any shape.
  • the edge 5 holds the production container 10 inside the collection container, and the inner surface of the bottom part of the collection container and the bottom part of the production container 10
  • the edge 5 can also be referred to as a holding portion, for example. The fixed distance will be described later.
  • the cell ink manufacturing kit of the present invention includes the cell ink manufacturing container of the present invention and a recovery container for recovering the cell mass, as described above.
  • the collection container has a bottomed cylindrical shape, and can accommodate the cell ink production container therein.
  • the production kit of the present invention is characterized by including the production container of the present invention, and other configurations and conditions are not particularly limited. According to the production kit of the present invention, for example, a cell ink capable of modeling a three-dimensional model to be described later can be manufactured. For example, the description of the cell ink of the present invention can be used for the production kit of the present invention.
  • FIG. 2A is a schematic exploded perspective view of the production kit of the present embodiment
  • FIG. 2B is a schematic perspective view of the production kit in which the production container is accommodated in the collection container.
  • (C) is a schematic cross-sectional view taken in the direction II-II in (B).
  • the manufacturing kit 20 of the present embodiment includes the manufacturing container 10 and the recovery container 11 of the embodiment.
  • the production container 10 is as described above.
  • the collection container 11 has a bottomed cylindrical shape and includes a holding portion 6 on the upper end side.
  • the cell ink is produced, for example, by introducing a culture solution and cells into the inside of the bottomed cylindrical container 1 of the production container 10, and removing the cells and the cell mass formed from the cells at the opening 3. It can be carried out by culturing, extracting the cultured cell mass from the inner surface side to the outer surface side of the opening 3 and collecting it in the collection container 11.
  • the manufacturing kit 20 of this embodiment includes the edge 5 and the holding part 6, the edge 5 and the holding part 6 are arbitrary structures, and may or may not be any one. May be included, or both may be included.
  • the formation material of the collection container 11 is not particularly limited.
  • the description of the formation material of the production container 10 can be cited by replacing “production container 10” with “recovery container 11”.
  • the forming material of the collection container 11 may be, for example, the same forming material as the manufacturing container 10 or a different forming material.
  • the number of the collection containers 11 is one, but the present invention is not limited to this, and may be plural.
  • the collection container 11 may be a bottomed cylindrical shape, and the shape is not particularly limited. Examples of the shape of the tube of the collection container 11 include a cylindrical shape and a rectangular tube shape.
  • the collection container 11 may have the same shape as the production container 10 or a different shape.
  • the number of collection containers 11 may be, for example, the same as or different from the number of containers 1 of the production container 11, but is preferably the former.
  • the collection container 11 is not particularly limited as long as the production container 10 can be accommodated therein, and its size is not particularly limited.
  • the volume, inner diameter, height, and the like of the collection container 11 can be appropriately set according to, for example, the production container 10 and the volume of the culture solution that can be cultured in the production container 10.
  • the collection container 11 may be, for example, a commercially available dish or a commercially available multiwell plate.
  • the dish include ⁇ 3 cm, ⁇ 5 cm, and ⁇ 10 cm dishes.
  • the multi-well plate include 6, 12, 24, 48, 96, and 384 well plates.
  • the production kit of the present invention can be used, for example, by arranging the production container 10 in the collection container 11.
  • the holding unit 6 is formed as the upper end of the recovery container 11, but the present invention is not limited to this, and the manufacturing container 10 is held inside the recovery container 11 and the recovery container 11
  • the inner surface of the bottom portion and the outer surface of the bottom portion 2 of the manufacturing container 10 may have any shape that can be arranged so as to maintain a certain distance.
  • the method by which the holding unit 6 holds the production container 10 is not particularly limited.
  • the collection container 11 may hold the production container 10 via the holding unit 6, or the collection container 11 may be the holding unit. 6 may be held in the production container 10 via
  • the fixed distance is not particularly limited, and is longer than 0 mm, for example.
  • the collection container 11 is a bottomed cylindrical container, but the present invention is not limited to this, and the collection container 11 is, for example, for a three-dimensional modeling apparatus that performs three-dimensional modeling by discharging ink. It may be an ink tank. In this case, the collection container 11 further includes an ink ejection unit. In the collection container 11, the ink ejecting portion may be disposed so as to eject ink, and the position thereof is not particularly limited. Examples of the position of the ink ejection unit in the collection container 11 include the bottom surface and the side surface of the collection container 11.
  • the production kit of the present invention may further include a lid that closes the opening of the production container 10 or the collection container 11.
  • the production kit of the present invention can prevent foreign matters from being mixed in, for example.
  • the collection container 11 is an ink tank of an ink jet 3D printer, for example, an ink cartridge (hereinafter also referred to as “ink cartridge”) of an ink jet 3D printer described later is mounted on the ink tank. )
  • the lid may be, for example, a lid that covers the production container 10 or the collection container 11, or a lid that is inscribed in the production container 10 or the collection container 11.
  • the method for producing a cell ink of the present invention includes a culture step of culturing cells using the cell ink production container of the present invention, and a recovery step of recovering the cultured cell mass.
  • cells are cultured in the opening of the cell ink production container in the presence of a culture solution, and in the collecting step, the cell mass led out of the cell ink production container from the opening. It is characterized by collect
  • the method for producing cell ink of the present invention for example, cell ink in which clogging of ink discharge ports when a solid is modeled by a three-dimensional modeling apparatus described later can be manufactured.
  • the description of the cell ink of the present invention can be used in the method for producing the cell ink of the present invention.
  • the type of the cell is not particularly limited and can be any cell.
  • the cell may be, for example, the same cell as a cell forming a cell mass obtained after culture (hereinafter also referred to as “target cell”) or a different cell.
  • the target cell can be appropriately determined according to, for example, the type of three-dimensional structure to be formed using the cell ink.
  • the target cells are, for example, parenchymal cells such as hepatocytes, sinusoidal wall cells, myofibroblasts, bile duct epithelial cells, connective tissue cells and the like. Etc.
  • examples of the cell include a cell that differentiates into the target cell.
  • the cells that differentiate into the target cells are not particularly limited, and examples thereof include pluripotent stem cells such as iPS cells and ES cells, stem cells such as somatic stem cells, and progenitor cells.
  • the origin of the cells is not particularly limited, and examples thereof include humans, non-human mammals, birds, reptiles, and seafood.
  • the culture medium is not particularly limited, and can be appropriately determined according to the type of the cell to be cultured.
  • the culture solution may further contain other components. Examples of the other components include a growth factor for growing the cell, a differentiation factor for differentiating the cell, and the like.
  • the growth factor and the differentiation factor are not particularly limited, and can be appropriately determined according to the type of the cell.
  • the type of cells constituting the cell mass is not particularly limited and can be appropriately adjusted depending on the conditions of the cells and culture.
  • the cell ink production container of the present invention is used, and the cells are cultured in the presence of a culture solution in the opening of the cell ink production container.
  • the cells and the culture solution are introduced into the bottomed cylindrical container of the production container from the upper opening of the production container and cultured.
  • the cells may be in a dispersed state or a cell mass at the start of the culture, but the former is preferable.
  • the type of the cells to be introduced may be, for example, one type or two or more types.
  • the cells may be cultured, for example, in the opening of the manufacturing container, in a bottomed cylindrical container of the manufacturing container, or in the manufacturing container.
  • the cells may be cultured in a hanging drop formed from the opening, or may be cultured in combination thereof. For example, the generation of a non-uniform cell mass due to contact between random cells can be suppressed, and more uniform. It is preferable to culture in the opening because a cell mass can be produced.
  • the production container may be centrifuged to introduce the cell into the opening.
  • the culture conditions in the culture are not particularly limited and can be appropriately set according to the type of the cell.
  • the culture temperature is, for example, 36 to 37 ° C.
  • the O 2 partial pressure is, for example, 1 to 21%
  • the CO 2 partial pressure is, for example, 5 to 6%.
  • the culture period in the culture is not particularly limited, and examples thereof include a period for forming a cell mass, and can be appropriately set according to the type of the cell.
  • the culture period is, for example, 12 hours to 30 days. Specifically, when the cells are hepatocytes, the culture period is, for example, 24 hours to 5 days.
  • the culture period for example, after the formation of the three-dimensional structure using the cell ink obtained by the production method of the present invention, adhesion between the cell masses is promoted, and cell death of the cells constituting the three-dimensional structure is reduced. Since it can suppress, it is preferable that it is the period until the said cell mass hold
  • the production container is accommodated in the collection container and the cells are cultured in the presence of the culture solution.
  • the culture solution may be introduced into the collection container in addition to the production container.
  • the cell mass collected in the collecting step can be used as a cell ink as it is, solution exchange for exchanging the culture solution with the cell ink solution prior to the collecting step. It is preferable to include a process.
  • the cell ink solution include a solution containing a component other than the cell mass in the cell ink.
  • Components other than the cell mass are not particularly limited, and examples thereof include lipids, the extracellular matrix, biomaterials, and stimulus-responsive molecules.
  • the stimulus-responsive molecule is a substance whose physical properties are changed by an external stimulus, for example.
  • the external stimulus may be, for example, a mechanical stimulus, an electromagnetic stimulus, or a chemical stimulus.
  • the mechanical stimulus include a physical stimulus, and specifically, a stimulus such as pressure and sound.
  • Examples of the electromagnetic stimulation include stimulation such as light and heat.
  • Examples of the change in physical properties include solidification, gelation, and liquefaction.
  • the cell ink solution may include, for example, a component other than one type of the cell mass, or may include two or more types of components other than the cell mass.
  • the solution constituting the cell ink solution is not particularly limited, and examples thereof include a phosphate buffer solution.
  • the culture medium and the cell ink solution can be exchanged by, for example, removing the culture liquid in the production container and introducing the cell ink solution into the production container.
  • the removal of the culture solution may be, for example, the removal of the culture solution in the bottomed cylindrical container of the production vessel, the removal of the culture solution in the opening, or the removal of both.
  • the removal and introduction are performed, for example, from the upper opening of the production container.
  • the collection step collects the cell mass led out of the cell ink production container from the opening.
  • the method for deriving the cell mass from the opening to the outside of the production container is not particularly limited. The derivation may be performed, for example, by spontaneously dropping a culture solution containing the cell mass from the inside of the production container to the outside of the production container, or applying pressure from the inside of the production container to the outside. Thus, it may be carried out by extruding the culture solution containing the cell mass from the opening, or by sucking out the culture solution containing the cell mass from the inside to the outside of the production container. May be.
  • the recovery step when a manufacturing kit including the manufacturing container is used, it is preferable to recover the cell mass derived from the opening to the recovery container outside the cell ink manufacturing container.
  • the recovery step for example, suppresses damage to the cell mass and can collect the cell mass, so that by introducing a recovery solution into the recovery container and bringing the opening and the recovery solution into contact with each other, More preferably, the cell mass led out to the collection container outside the cell ink production container is collected from the opening.
  • the recovered solution include the cell ink solution.
  • the collection of the cell mass can be carried out, for example, by housing the production container inside the collection container and leading the cell mass out of the production container through the opening.
  • the collection step it is preferable that in the collection step, the cell ink production container is removed from the collection container and the cell mass in the collection container is collected.
  • the cell mass recovered in the recovery step may be used as cell ink, or a solution obtained by dispersing the recovered cell mass in the cell ink solution may be used as cell ink.
  • the cell ink may include, for example, one type of cell mass or two or more types of cell mass.
  • the cell ink of the present invention is obtained by the method for producing the cell ink of the present invention.
  • the cell ink of the present invention is obtained by the method for producing the cell ink of the present invention, and other configurations and conditions are not particularly limited.
  • the cell ink of the present invention for example, when a three-dimensional model is modeled by an ink jet 3D printer described later, clogging of the ink discharge port of the ink jet 3D printer can be reduced.
  • the effect of the cell ink described above can be obtained.
  • the description of the cell ink of the present invention can be used for the cell ink of the present invention.
  • an ink cartridge for a three-dimensional modeling apparatus that performs three-dimensional modeling by discharging the ink of the present invention includes the cell ink of the present invention.
  • the ink cartridge of the present invention is characterized by containing the cell ink of the present invention, and other configurations and conditions are not particularly limited. According to the ink cartridge of the present invention, it is possible to easily model a three-dimensional model using, for example, an ink jet 3D printer.
  • the description of the cell ink of the present invention can be cited.
  • the ink cartridge of the present invention may contain, for example, one type of cell ink or two or more types of cell ink.
  • the ink cartridge may contain, for example, the cell ink in the same container or not mixed, or may be contained in a separate container.
  • the ink cartridge of the present invention can also be referred to as an ink set, for example.
  • the method for manufacturing a three-dimensional object of the present invention includes a three-dimensional object forming process using cell ink, and the three-dimensional object forming process performs three-dimensional object formation by discharging ink based on the three-dimensional data of the object to be formed.
  • the cell ink is ejected from the ink discharge port of the three-dimensional modeling apparatus to form a three-dimensional modeled object, and the cell ink is obtained by the cell ink of the present invention and the method for producing the cell ink of the present invention. It is at least one of ink.
  • the method for producing a three-dimensional structure of the present invention is characterized in that a three-dimensional structure is formed using at least one of the cell ink of the present invention and the cell ink obtained by the method of manufacturing the cell ink of the present invention.
  • the other steps and conditions are not particularly limited.
  • a three-dimensional structure formed from cells such as foods and transplanted organs can be produced.
  • the manufacturing method of the three-dimensional molded item of this invention can also model the three-dimensional molded item which can supply a nutrient, oxygen, etc. to an inside by modeling a blood vessel etc., for example.
  • the manufacturing method of the three-dimensional molded item of this invention compared with the case where an organ etc. are modeled by culture
  • the description of the cell ink of the present invention can be used in the method for producing a three-dimensional structure of the present invention.
  • the cell ink used in the method for producing a three-dimensional structure of the present invention includes the cell mass, for example, when forming a three-dimensional structure of the same size, the cell ink is shorter than a cell ink including a single cell. Can be shaped in time.
  • the manufacturing method of the three-dimensional modeled object of the present invention for example, by using the cell ink, the three-dimensional modeled object can be modeled in a shorter time. Therefore, the cell death of the cells constituting the modeled three-dimensional modeled object Can be suppressed. Furthermore, in the cell ink, for example, the density of cell adhesion molecules on the cell surface of the cells constituting the cell mass is not impaired. For this reason, according to the manufacturing method of the three-dimensional molded item of this invention, when food is modeled as the said three-dimensional molded item using the said cell ink, the food with high freshness and the original taste can be modeled. Further, according to the method for manufacturing a three-dimensional structure of the present invention, for example, when an organ is formed as the three-dimensional structure using the cell ink, an organ in which the function of the organ is maintained can be formed.
  • Examples of the three-dimensional data include data that defines the three-dimensional structure of the target three-dimensional object.
  • Examples of the three-dimensional data include three-dimensional CT (computed tomography) images, three-dimensional MRI (magnetic resonance tomography) images, three-dimensional PET (positron tomography) tomography images, and three-dimensional PET-CT images.
  • the three-dimensional data preferably includes information on a tissue constituting the modeling target because, for example, an organ having a structure closer to a living organ or the like can be modeled.
  • the information include information on structure, volume, density, tissue state, and the like.
  • the tissue information include information on tissues such as epithelial tissue, connective tissue, nerve tissue, vascular tissue, adipose tissue, and muscle tissue.
  • the organization information may be one type or two or more types.
  • the three-dimensional model is not particularly limited as long as it is a three-dimensional model composed of the cells.
  • Examples of the three-dimensional model include foods and organs.
  • the food is not particularly limited, and examples thereof include any food such as liver, tro, sirloin, fin, and loin.
  • the organ is not particularly limited, and examples thereof include liver, heart, kidney, pancreas, and lung.
  • the organ may be, for example, a part or the whole of the organ. Examples of the organ include an organ.
  • the type of the cell ink to be used is not particularly limited and can be appropriately determined according to, for example, the three-dimensional model.
  • the number of the cell inks to be used is not particularly limited, and can be appropriately set according to, for example, the number of types of cells constituting the modeling object.
  • the number of the cell inks used may be, for example, one type or two or more types. In the latter case, the types of cells constituting the cell mass included in each cell ink may be the same or different, for example.
  • the size of the cell mass which each cell ink contains may be the same, for example, and may differ.
  • the method for manufacturing a three-dimensional structure of the present invention may further include a cell ink manufacturing process for manufacturing the cell ink.
  • the manufacturing process of the cell ink can be performed by, for example, the cell ink manufacturing method of the present invention.
  • the cell ink obtained by the said manufacturing process is used, for example.
  • the manufacturing method of the three-dimensional modeled object of the present invention may further include a three-dimensional data acquisition step of acquiring the three-dimensional data of the modeling target prior to the three-dimensional modeling step.
  • the three-dimensional data is acquired by, for example, the inkjet 3D printer.
  • the three-dimensional data acquired in the three-dimensional data acquisition step is not particularly limited, and can be appropriately determined according to the modeling object. Specifically, when the three-dimensional object is a modeling object including a blood vessel, in the three-dimensional data acquisition step, for example, three-dimensional data including blood vessel tissue information of the modeling object is acquired.
  • the method for acquiring the three-dimensional data is not particularly limited.
  • the three-dimensional data may be acquired from a storage medium in which the three-dimensional data is stored, or the three-dimensional data may be acquired by measuring the modeling object. Good.
  • the manufacturing method of the three-dimensional structure of the present invention includes, for example, a tomographic image acquisition step of acquiring a tomographic image of the modeling object and a three-dimensional data generation step of generating the three-dimensional data from the tomographic image.
  • the acquisition of the tomographic image can be acquired, for example, by measuring the modeling object with a tomographic image acquisition device such as CT, MRI, or PET.
  • the three-dimensional data can be produced, for example, from the measurement data obtained by the tomographic image acquisition apparatus, from the three-dimensional CT image, the three-dimensional MRI image, the three-dimensional PET image, the three-dimensional PET-CT image, etc. Can be obtained by creating the three-dimensional data.
  • the modeling object is an organ of a human or non-human animal and the stereoscopic data is stereoscopic data including vascular tissue information, for example, a known angiography method for the human or non-human animal.
  • two or more tomographic images of the modeling object are acquired.
  • the three-dimensional data can be acquired by performing, for example, a known volume data conversion process on the two or more tomographic images.
  • the cellular ink is ejected from an ink discharge port of a three-dimensional modeling apparatus that performs ink three-dimensional modeling by discharging ink, thereby modeling a three-dimensional model.
  • the ink jet 3D printer is, for example, an ink jet 3D printer that includes an ink container and an ink discharge unit, and discharges ink stored in the ink container by the ink discharge unit.
  • the ink cartridge of the present invention is accommodated in the ink accommodating portion.
  • the modeling of the three-dimensional model can be appropriately performed according to, for example, the type of the ink jet 3D printer.
  • the three-dimensional data is input to the ink jet 3D printer and the model is modeled with default settings.
  • the ink jet 3D printer may be, for example, an ink jet 3D printer capable of ejecting one type of ink or an ink jet 3D printer capable of ejecting two or more types of ink.
  • the cell ink for example, a cell ink obtained by the cell ink production method of the present invention may be used instead of the cell ink of the present invention.
  • the ink jet 3D printer may model the three-dimensional model by discharging other ink from the ink discharge port in addition to the cell ink.
  • the other ink include an ink containing a component other than the cell mass. Specifically, an ink containing a gel or the like, an ink containing a seasoning, an ink containing a water-soluble polymer, the stimulus responsiveness, and the like. Examples thereof include ink containing molecules and ink containing the extracellular matrix.
  • a complex organ including a plurality of tissues can be modeled. Therefore, the modeling object includes a plurality of tissues, and the cell ink corresponds to each tissue.
  • 3D modeling apparatus for performing 3D modeling by discharging ink based on 3D data of a modeling object including information on the plurality of tissues using the plurality of cell inks in the 3D modeling process Preferably, the cellular ink is discharged from the ink discharge port to form a three-dimensional structure including a plurality of tissues.
  • the modeling object is a modeling object including a vascular tissue, a fat tissue, and a muscle tissue, for example, it can be performed as follows. Note that these are examples and do not limit the present invention.
  • a cell ink containing a cell mass composed of cells contained in each tissue is produced by the cell ink production method of the present invention.
  • the cell ink corresponding to the vascular tissue includes, for example, vascular endothelial cells.
  • the cell ink corresponding to the fat tissue includes, for example, fat cells.
  • the ink corresponding to the muscle tissue includes, for example, muscle cells.
  • the size of each cell mass is set according to, for example, the proportion of each tissue in the modeling object.
  • the said cell ink is accommodated in the ink cartridge accommodated in the said inkjet type 3D printer.
  • the ink cartridge containing each cell ink is accommodated in the ink accommodating portion of the ink jet 3D printer. Further, the ink jet 3D printer acquires three-dimensional data including information on the vascular tissue, the fat tissue, and the muscle tissue. And it sets so that the cell ink corresponding to each said tissue may be discharged to the position corresponding to each said tissue. In addition, when the said modeling target object has a space
  • the inkjet 3D printer ejects each cell ink to a position corresponding to each tissue, thereby modeling the three-dimensional structure including each tissue.
  • the cell ink containing a vascular endothelial cell etc. is ejected as an example of the cell ink corresponding to the vascular tissue
  • differentiation into the vascular tissue may be shaped using a cell ink containing progenitor cells, an ink containing the stimulus-responsive molecule, an ink containing the extracellular matrix, or the like.
  • the three-dimensional model can be modeled.
  • the three-dimensional object of the present invention is obtained by the method for manufacturing a three-dimensional object of the present invention.
  • the three-dimensional modeled object of the present invention is obtained by the method for manufacturing a three-dimensional modeled object of the present invention, and other configurations and conditions are not particularly limited.
  • the description of the cell ink of the present invention can be used for the three-dimensional modeled object of the present invention.
  • this invention it is possible to produce a cell ink capable of modeling a three-dimensional model. For this reason, this invention is very useful in the medical field, the foodstuff field, etc., for example.

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Abstract

Provided are: a cell ink capable of shaping a sterically formed object; a cell ink production container capable of producing the cell ink; a cell ink production kit; a cell ink production method; an ink cartridge; a sterically formed object production method; and a sterically formed object. The cell ink according to the present invention is characterized by containing a cell aggregation. In the cell ink, the cell aggregation is preferably a homogeneous cell aggregation, and, more specifically, in the cell aggregation contained in the cell ink, 90% or more of the cell aggregation further preferably has a diameter within a range of ±30% with respect to a predetermined diameter.

Description

細胞インク、細胞インクの製造容器、細胞インクの製造キット、細胞インクの製造方法、インクカートリッジ、立体造形物の製造方法、および立体造形物Cell ink, cell ink manufacturing container, cell ink manufacturing kit, cell ink manufacturing method, ink cartridge, three-dimensional object manufacturing method, and three-dimensional object
　本発明は、細胞インク、細胞インクの製造容器、細胞インクの製造キット、細胞インクの製造方法、インクカートリッジ、立体造形物の製造方法、および立体造形物に関する。 The present invention relates to a cell ink, a cell ink production container, a cell ink production kit, a cell ink production method, an ink cartridge, a method for producing a three-dimensional object, and a three-dimensional object.
　ｉＰＳ細胞、ＥＳ細胞等の多能性細胞を三次元的に培養し、臓器等を作製することが試みられている（特許文献１）。 An attempt has been made to produce organs and the like by three-dimensionally culturing pluripotent cells such as iPS cells and ES cells (Patent Document 1).
　しかしながら、臓器等の立体造形物を細胞培養により作製する場合、作製した立体造形物の内部への栄養、酸素等の供給の問題から、例えば、実際の臓器と同等の大きさの臓器を作製することが困難という問題があった。 However, when a three-dimensional object such as an organ is produced by cell culture, an organ having the same size as the actual organ is produced, for example, due to the supply of nutrients and oxygen to the inside of the produced three-dimensional object. There was a problem that it was difficult.
特開２００８－０４９１４６号公報JP 2008-049146 A
　そこで、本発明は、立体造形物の造形が可能な細胞インク、細胞インクを製造可能な細胞インクの製造容器、細胞インクの製造キット、細胞インクの製造方法、インクカートリッジ、立体造形物の製造方法、および立体造形物を提供する。 Accordingly, the present invention relates to a cell ink capable of modeling a three-dimensional structure, a cell ink manufacturing container capable of manufacturing a cell ink, a cell ink manufacturing kit, a method of manufacturing a cell ink, an ink cartridge, and a method of manufacturing a three-dimensional structure. And a three-dimensionally shaped object.
　本発明の細胞インクは、細胞塊を含むことを特徴とする。 The cell ink of the present invention is characterized by containing a cell mass.
　本発明の細胞インクの製造容器は、有底筒状の容器を含み、
前記容器の底部は、１以上の開口部を含み、
前記開口部は、細胞塊を培養するための培養室であり、
前記底部の外面側における前記開口部の径の大きさが、インクを吐出して立体造形を行う立体造形装置のインク吐出口の径の大きさ以下であることを特徴とする。 The cell ink production container of the present invention includes a bottomed cylindrical container,
The bottom of the container includes one or more openings;
The opening is a culture chamber for culturing a cell mass,
The size of the diameter of the opening on the outer surface side of the bottom is not more than the size of the diameter of the ink discharge port of the three-dimensional modeling apparatus that performs three-dimensional modeling by discharging ink.
　本発明の細胞インクの製造キットは、前記本発明の細胞インクの製造容器と、細胞塊を回収するための回収容器とを含み、
前記回収容器は、有底筒状であり、内部に前記細胞インクの製造容器を収容可能であることを特徴とする。 The cell ink production kit of the present invention includes the cell ink production container of the present invention and a collection container for collecting the cell mass,
The collection container has a bottomed cylindrical shape, and can accommodate the cell ink production container therein.
　本発明の細胞インクの製造方法は、前記本発明の細胞インクの製造容器を用いて細胞を培養する培養工程、および
培養された細胞塊を回収する回収工程を含み、
前記培養工程において、培養液の存在下、前記細胞インクの製造容器の前記開口部において、細胞を培養し、
前記回収工程において、前記開口部から、前記細胞インクの製造容器の外部に導出された細胞塊を回収することを特徴とする。 The method for producing a cell ink of the present invention includes a culture step of culturing cells using the cell ink production container of the present invention, and a recovery step of collecting the cultured cell mass,
In the culturing step, cells are cultured in the opening of the cell ink production container in the presence of a culture solution,
In the collecting step, the cell mass led out of the cell ink production container is collected from the opening.
　本発明の細胞インクは、前記本発明の細胞インクの製造方法により得られることを特徴とする。 The cell ink of the present invention is obtained by the method for producing the cell ink of the present invention.
　本発明のインクを吐出して立体造形を行う立体造形装置用のインクカートリッジは、前記本発明の細胞インクを含むことを特徴とする。 An ink cartridge for a three-dimensional modeling apparatus that performs three-dimensional modeling by discharging the ink of the present invention includes the cell ink of the present invention.
　本発明の立体造形物の製造方法は、細胞インクを用いる立体造形工程を含み、
前記立体造形工程が、造形対象物の立体データに基づき、インクを吐出して立体造形を行う立体造形装置のインク吐出口から、前記細胞インクを吐出し、立体造形物を造形し、
前記細胞インクが、前記本発明の細胞インク、および前記本発明の細胞インクの製造方法により得られた細胞インクの少なくとも一方であることを特徴とする。 The method for producing a three-dimensional structure of the present invention includes a three-dimensional modeling process using cell ink,
The three-dimensional modeling step is based on the three-dimensional data of the modeling object, and discharges the cell ink from the ink discharge port of the three-dimensional modeling apparatus that performs ink three-dimensional modeling by discharging ink, and models the three-dimensional modeling object,
The cell ink is at least one of the cell ink of the present invention and the cell ink obtained by the method for producing the cell ink of the present invention.
　本発明の立体造形物は、前記本発明の立体造形物の製造方法により得られることを特徴とする。 The three-dimensional structure of the present invention is obtained by the method for manufacturing a three-dimensional structure of the present invention.
　本発明によれば、立体造形物の造形が可能な細胞インクを提供可能である。 According to the present invention, it is possible to provide a cell ink capable of modeling a three-dimensional model.
図１（Ａ）は、本発明の細胞インクの製造容器の一例を示す模式透視斜視図であり、図１（Ｂ）は、図１（Ａ）において、Ｉ－Ｉ方向からみた模式断面図である。FIG. 1 (A) is a schematic perspective view showing an example of the cell ink production container of the present invention, and FIG. 1 (B) is a schematic cross-sectional view as seen from the II direction in FIG. 1 (A). is there. 図２（Ａ）は、本発明の細胞インクの製造キットの一例を示す模式分解斜視図であり、図２（Ｂ）は、前記製造キットにおいて、前記製造容器を前記回収容器に収容した状態の模式透視斜視図であり、図２（Ｃ）は、図２（Ｂ）において、II－II方向からみた模式断面図である。FIG. 2 (A) is a schematic exploded perspective view showing an example of the cell ink production kit of the present invention, and FIG. 2 (B) shows a state in which the production container is housed in the collection container in the production kit. FIG. 2C is a schematic perspective view, and FIG. 2C is a schematic cross-sectional view as seen from the II-II direction in FIG. 2B.
　つぎに、本発明の実施形態について説明する。なお、本発明は、下記の実施形態によって何ら限定および制限されない。なお、以下の図１および図２において、同一部分には、同一符号を付している。また、各実施形態の説明は、特に言及がない限り、互いの説明を援用できる。さらに、各実施形態の構成は、特に言及がない限り、組合せ可能である。 Next, an embodiment of the present invention will be described. In addition, this invention is not limited and restrict | limited at all by the following embodiment. In addition, in the following FIG. 1 and FIG. 2, the same code | symbol is attached | subjected to the same part. Moreover, the description of each embodiment can use each other's description unless there is particular mention. Further, the configurations of the embodiments can be combined unless otherwise specified.
　以下の説明において、「下方向」は、前記有底筒状の容器の底面方向を意味し、「上方向」は、前記有底筒状の容器の底面方向の反対方向、すなわち開口方向を意味し、上下方向の長さは、高さともいう。また、「内側および内部」は、前記有底筒状の容器の底面および筒状の側壁に囲われた領域を意味し、「外側および外部」は、前記有底筒状の容器の底面および筒状の側壁に囲われない領域を意味する。 In the following description, “downward direction” means the bottom surface direction of the bottomed cylindrical container, and “upward direction” means the direction opposite to the bottom surface direction of the bottomed cylindrical container, that is, the opening direction. The vertical length is also called height. Further, “inside and inside” means a region surrounded by a bottom surface and a cylindrical side wall of the bottomed cylindrical container, and “outside and outside” means a bottom surface and a cylinder of the bottomed cylindrical container. Means a region not surrounded by a side wall.
＜細胞インク＞
　本発明の細胞インクは、前述のように、細胞塊を含むことを特徴とする。本発明の細胞インクは、前記細胞塊を含むことが特徴であり、その他の構成および条件は、特に制限されない。本発明の細胞インクは、前記細胞塊を含むため、例えば、同じ大きさの立体造形物を造形する場合、単細胞を含む細胞インクと比較して、より短時間で造形できる。また、本発明の細胞インクによれば、例えば、より短時間で立体造形物を造形できることから、造形した前記立体造形物を構成する細胞の細胞死を抑制できる。さらに、本発明の細胞インクは、例えば、前記細胞塊を構成する細胞の細胞表面における細胞接着分子の密度が損なわれていない。このため、本発明の細胞インクによれば、例えば、前記立体造形物として食品を造形した場合、鮮度が高く、本来の食味に近い食品が造形できる。また、本発明の細胞インクによれば、例えば、前記立体造形物として臓器を造形した場合、前記臓器の機能が維持された臓器が造形できる。また、本発明の細胞インクによれば、例えば、後述するインクを吐出して立体造形を行なう立体造形装置（以下、「インクジェット式３Ｄプリンター」ともいう。）により立体造形物を造形した際に、前記インクジェット式３Ｄプリンターのインク吐出口の目詰まりを低減できる。 <Cellular ink>
As described above, the cell ink of the present invention includes a cell mass. The cell ink of the present invention is characterized by containing the cell mass, and other configurations and conditions are not particularly limited. Since the cell ink of the present invention includes the cell mass, for example, when modeling a three-dimensional model having the same size, the cell ink can be modeled in a shorter time than a cell ink including a single cell. In addition, according to the cell ink of the present invention, for example, since a three-dimensional model can be modeled in a shorter time, cell death of cells constituting the modeled three-dimensional model can be suppressed. Furthermore, in the cell ink of the present invention, for example, the density of cell adhesion molecules on the cell surface of the cells constituting the cell mass is not impaired. For this reason, according to the cell ink of the present invention, for example, when a food is modeled as the three-dimensional model, a food with high freshness and close to the original taste can be modeled. Moreover, according to the cell ink of the present invention, for example, when an organ is modeled as the three-dimensional modeled object, an organ in which the organ function is maintained can be modeled. Further, according to the cell ink of the present invention, for example, when a three-dimensional object is modeled by a three-dimensional modeling apparatus (hereinafter, also referred to as “inkjet 3D printer”) that performs three-dimensional modeling by discharging ink described later, It is possible to reduce clogging of the ink discharge port of the ink jet 3D printer.
　本発明の細胞インクにおいて、前記細胞塊を構成する細胞の種類は、特に制限されず、例えば、前記細胞インクを使用して造形する立体造形物の種類に応じて、適宜決定できる。前記細胞は、例えば、後述する目的細胞と同じ細胞または異なる細胞があげられる。前記立体造形物が肝臓である場合、前記細胞は、例えば、肝細胞等の実質細胞、類洞壁細胞、筋線維芽細胞、胆管上皮細胞、結合組織細胞等の非実質細胞等があげられる。前記細胞塊は、複数の前記細胞から形成される凝集体を意味し、例えば、スフェロイドということもできる。前記細胞塊は、例えば、１種類の細胞から構成されてもよいし、２種類以上の細胞から構成されてもよい。前記細胞の由来は、特に制限されず、例えば、ヒト、非ヒト哺乳類、鳥類、は虫類、魚介類等があげられる。 In the cell ink of the present invention, the type of cells constituting the cell mass is not particularly limited, and can be appropriately determined according to, for example, the type of three-dimensional modeled object to be modeled using the cell ink. Examples of the cell include the same cell as the target cell described later or a different cell. When the three-dimensional model is a liver, examples of the cells include parenchymal cells such as hepatocytes, sinusoidal wall cells, myofibroblasts, bile duct epithelial cells, and connective tissue cells. The cell mass means an aggregate formed from a plurality of the cells, and can also be referred to as, for example, a spheroid. The cell mass may be composed of, for example, one type of cell or may be composed of two or more types of cells. The origin of the cell is not particularly limited, and examples thereof include humans, non-human mammals, birds, reptiles, and seafood.
　前記細胞塊を構成する細胞の数は、特に制限されず、例えば、１０～１００００個であり、前記細胞塊内部の細胞の細胞死を抑制できることから、好ましくは、１０～５０００個である。 The number of cells constituting the cell mass is not particularly limited, and is, for example, 10 to 10,000, and is preferably 10 to 5000 because cell death of cells inside the cell mass can be suppressed.
　前記細胞塊の大きさは、特に制限されず、例えば、その径が、２０～９００μｍ、５０～５００μｍである。前記細胞塊の径は、例えば、短径でもよいし、長径でもよいし、平均径でもよいが、好ましくは、長径である。前記細胞塊の径は、例えば、インクジェット式３Ｄプリンターを用いて、立体造形物を造形した際に、前記インクジェット式３Ｄプリンターのインク吐出口の目詰まりをより抑制できることから、好ましくは、前記インクジェット式３Ｄプリンターのインク吐出口の径（例えば、直径。以下、同様。）の大きさ以下であり、より好ましくは、前記インクジェット式３Ｄプリンターのインク吐出口の径に対して１／４～１倍、１／３～１／１．２倍の範囲の長さである。前記インク吐出口の径は、例えば、短径でもよいし、長径でもよいし、平均径でもよいが、好ましくは、短径である。前記インクジェット式３Ｄプリンターのインク吐出口の径は、例えば、２０～４０００μｍ、５０～２０００μｍである。 The size of the cell mass is not particularly limited, and for example, the diameter is 20 to 900 μm, 50 to 500 μm. The diameter of the cell mass may be, for example, a short diameter, a long diameter, or an average diameter, but is preferably a long diameter. For example, when the three-dimensional model is formed using an ink jet 3D printer, the diameter of the cell mass can more effectively suppress clogging of the ink discharge port of the ink jet 3D printer. The diameter of the ink discharge port of the 3D printer (for example, the diameter, the same applies hereinafter) or less, more preferably 1/4 to 1 times the diameter of the ink discharge port of the ink jet 3D printer, The length is in the range of 1/3 to 1 / 1.2 times. The diameter of the ink discharge port may be, for example, a short diameter, a long diameter, or an average diameter, but is preferably a short diameter. The diameter of the ink discharge port of the ink jet 3D printer is, for example, 20 to 4000 μm, 50 to 2000 μm.
　本発明の細胞インクにおいて、例えば、前記インクジェット式３Ｄプリンターのインク吐出口の目詰まりをより抑制できることから、前記細胞塊は、均一な細胞塊で有ることが好ましく、目詰まりをさらに抑制できることから、前記細胞インクに含まれる細胞塊において、９０％以上、９５％以上、または９９％以上の細胞塊の径（Ｄｃ）が、所定径（Ｄｐ）に対して、±３０％（０．７×Ｄｐ≦Ｄｃ≦１．３×Ｄｐ）、±２０％（０．８×Ｄｐ≦Ｄｃ≦１．２×Ｄｐ）、または±１０％（０．９×Ｄｐ≦Ｄｃ≦１．１×Ｄｐ）の範囲であることが好ましい。前記所定径は、例えば、前記インクジェット式３Ｄプリンターのインク吐出口の径に応じて、適宜設定できる。前記所定径は、例えば、２０～４０００μｍ、５０～２０００μｍである。前記細胞塊の径は、目詰まりをさらに抑制できることから、前記細胞インクに含まれる細胞塊において、８５％以上、９０％以上、９５％以上、９６％以上、９７％以上、９８％以上、９９％以上、または１００％の細胞塊が、前記インクジェット式３Ｄプリンターのインク吐出口の径に対して、９５％以下、９０％以下、または８５％以下の径を有することが好ましく、１００％の細胞塊が、前記インクジェット式３Ｄプリンターのインク吐出口の径に対して、８５％以下の径を有することがより好ましい。 In the cell ink of the present invention, for example, since the clogging of the ink discharge port of the ink jet 3D printer can be further suppressed, the cell mass is preferably a uniform cell mass, and can further suppress clogging. In the cell mass contained in the cell ink, the diameter (D c ) of 90% or more, 95% or more, or 99% or more of the cell mass is ± 30% (0.7% relative to the predetermined diameter (D p ). × D p ≦ D c ≦ 1.3 × D p ), ± 20% (0.8 × D p ≦ D c ≦ 1.2 × D p ), or ± 10% (0.9 × D p ≦ D c ≦ 1.1 × D p ). The predetermined diameter can be appropriately set according to, for example, the diameter of the ink discharge port of the ink jet 3D printer. The predetermined diameter is, for example, 20 to 4000 μm, 50 to 2000 μm. Since the diameter of the cell mass can further suppress clogging, in the cell mass contained in the cell ink, 85% or more, 90% or more, 95% or more, 96% or more, 97% or more, 98% or more, 99 % Or 100% of the cell mass preferably has a diameter of 95% or less, 90% or less, or 85% or less with respect to the diameter of the ink discharge port of the ink jet 3D printer, and 100% of cells It is more preferable that the lump has a diameter of 85% or less with respect to the diameter of the ink discharge port of the ink jet 3D printer.
　前記細胞塊は、例えば、前記立体造形物の造形後、前記細胞塊間の接着が促進され、前記立体造形物を構成する細胞の細胞死を抑制できることから、前記細胞塊を構成する細胞の細胞表面における細胞接着分子の密度が損なわれていないことが好ましい。前記細胞接着分子は、例えば、インテグリン、カドヘリン等があげられる。前記細胞接着分子の密度は、例えば、１種類の細胞接着分子の密度でもよいし、２種類以上の細胞接着分子の密度の平均の密度でもよい。 The cell mass is, for example, a cell of a cell constituting the cell mass because adhesion between the cell masses is promoted after shaping the three-dimensional modeled object and cell death of the cells constituting the three-dimensional modeled product can be suppressed. It is preferred that the density of cell adhesion molecules on the surface is not impaired. Examples of the cell adhesion molecule include integrin, cadherin and the like. The density of the cell adhesion molecule may be, for example, the density of one type of cell adhesion molecule or the average density of two or more types of cell adhesion molecules.
　前記細胞塊は、例えば、前記立体造形物の造形後、前記細胞塊間の接着が促進され、前記立体造形物を構成する細胞の細胞死を抑制できることから、前記細胞塊の周囲に細胞外マトリックスを保持していることが好ましい。前記細胞外マトリックスは、例えば、コラーゲン、プロテオグリカン、フィブロネクチン、ラミニン等のタンパク質、グリコサミノグリカン等の糖等があげられる。 The cell mass is, for example, after the modeling of the three-dimensional modeled object, the adhesion between the cell masses is promoted, and the cell death of the cells constituting the three-dimensional modeled product can be suppressed. Is preferably maintained. Examples of the extracellular matrix include proteins such as collagen, proteoglycan, fibronectin and laminin, and sugars such as glycosaminoglycan.
　本発明の細胞インクは、例えば、その他の成分を含んでもよい。前記その他の成分は、例えば、前記細胞塊を分散させる溶媒があげられ、具体的には、後述する細胞インク用溶液等があげられる。 The cell ink of the present invention may contain other components, for example. Examples of the other components include a solvent for dispersing the cell mass, and specific examples include a cell ink solution described later.
　本発明の細胞インクは、例えば、後述する細胞インクの製造方法により製造できる。 The cell ink of the present invention can be produced by, for example, a cell ink production method described later.
＜細胞インクの製造容器＞
　本発明の細胞インクの製造容器（以下、「製造容器」ともいう。）は、前述のように、有底筒状の容器を含み、前記容器の底部は、１以上の開口部を含み、前記開口部は、細胞塊を培養するための培養室であり、前記底部の外面側における前記開口部の径の大きさが、インクを吐出して立体造形を行う立体造形装置のインク吐出口の径の大きさ以下であることを特徴とする。本発明の製造容器は、前記製造容器において、前記底部の外面側における前記開口部の径の大きさが、インクを吐出して立体造形を行う立体造形装置のインク吐出口の径の大きさ以下であることが特徴であり、その他の構成および条件は、特に制限されない。本発明の製造容器によれば、例えば、後述するように立体造形物の造形が可能な細胞インクを製造可能である。また、本発明の製造容器は、前記開口部の径の大きさが、前記立体造形装置のインク吐出口の径の大きさ以下である。このため、前記開口部から前記製造容器の外部に導出された細胞塊の径の大きさが、例えば、前記立体造形装置のインク吐出口の径の大きさ以下であり、且つ細胞インクに含まれる細胞塊の径の大きさの均一性が高い。したがって、前記細胞塊を含む細胞インクで立体造形を行った際に、例えば、前記立体造形装置のインク吐出口において、前記細胞インクの目詰まりを低減できる。このため、本発明の細胞インクの製造容器によれば、例えば、前記立体造形装置により立体を造形した際のインク吐出口の目詰まりが低減された細胞インクを製造できる。本発明の製造容器は、例えば、前記本発明の細胞インク等の説明を援用できる。 <Cellular ink manufacturing container>
The cell ink production container of the present invention (hereinafter also referred to as “production container”) includes a bottomed cylindrical container as described above, and the bottom of the container includes one or more openings, The opening is a culture chamber for culturing a cell mass, and the size of the diameter of the opening on the outer surface side of the bottom is a diameter of an ink discharge port of a three-dimensional modeling apparatus that performs three-dimensional modeling by discharging ink. The size is less than or equal to. In the manufacturing container of the present invention, in the manufacturing container, the diameter of the opening on the outer surface side of the bottom is equal to or smaller than the diameter of the ink discharge port of the three-dimensional modeling apparatus that performs three-dimensional modeling by discharging ink. The other features and conditions are not particularly limited. According to the production container of the present invention, for example, a cell ink capable of modeling a three-dimensional model can be manufactured as described later. In the manufacturing container of the present invention, the diameter of the opening is equal to or less than the diameter of the ink discharge port of the three-dimensional modeling apparatus. For this reason, the size of the diameter of the cell mass derived from the opening to the outside of the manufacturing container is, for example, not more than the size of the diameter of the ink discharge port of the three-dimensional modeling apparatus, and is included in the cell ink. The uniformity of the size of the cell mass is high. Therefore, when three-dimensional modeling is performed with the cell ink containing the cell mass, for example, clogging of the cell ink can be reduced at the ink discharge port of the three-dimensional modeling apparatus. For this reason, according to the cell ink manufacturing container of the present invention, for example, it is possible to manufacture a cell ink in which clogging of ink discharge ports when a solid is modeled by the three-dimensional modeling apparatus is reduced. For example, the description of the cell ink of the present invention can be used for the production container of the present invention.
　本発明の製造容器の一例について、図面を参照し説明する。また、図面においては、説明の便宜上、各部の構造は適宜簡略化して示す場合があり、各部の寸法比等は、実際とは異なり、模式的に示す場合がある。 An example of the production container of the present invention will be described with reference to the drawings. In the drawings, for convenience of explanation, the structure of each part may be simplified as appropriate, and the dimensional ratio of each part may be schematically shown, unlike the actual case.
　図１（Ａ）は、本実施形態の製造容器の模式透視斜視図であり、（Ｂ）は、（Ａ）において、Ｉ－Ｉ方向からみた模式断面図である。図１（Ａ）および（Ｂ）に示すように、本実施形態の製造容器１０は、有底筒状の容器１を含み、容器１は、底部２と側壁４と縁５とを含む。製造容器１０において、底部２は、側壁４の下側端に、縁５は、側壁４の上側端に、側壁４と一体となるように形成されている。縁５は、側壁４の外方向に突出しており、例えば、製造容器１０を把持する把持部となる。また、容器１の底部２は、複数の開口部３を含み、開口部３は、底部２の内面側から外面側に向かってテーパー状である。製造容器１０において、細胞インクの製造は、例えば、有底筒状の容器１の内側に培養液および細胞を導入し、開口部３で細胞および前記細胞から形成された細胞塊を培養することにより実施できる。このため、開口部３は、前記細胞塊を培養するための培養室ということもできる。前記細胞および前記細胞塊については、後述する。なお、本実施形態の製造容器１０は、縁５を含むが、縁５は、任意の構成であり、あってもよいし、なくてもよい。 FIG. 1A is a schematic perspective view of the production container of the present embodiment, and FIG. 1B is a schematic cross-sectional view as viewed from the II direction in FIG. As shown in FIGS. 1A and 1B, the manufacturing container 10 of the present embodiment includes a bottomed cylindrical container 1, and the container 1 includes a bottom portion 2, a side wall 4, and an edge 5. In the production container 10, the bottom 2 is formed at the lower end of the side wall 4, and the edge 5 is formed at the upper end of the side wall 4 so as to be integrated with the side wall 4. The edge 5 protrudes outward of the side wall 4 and becomes, for example, a gripping part that grips the manufacturing container 10. The bottom 2 of the container 1 includes a plurality of openings 3, and the openings 3 are tapered from the inner surface side to the outer surface side of the bottom portion 2. In the production container 10, the cell ink is produced, for example, by introducing a culture solution and cells into the inside of the bottomed cylindrical container 1 and culturing the cells and the cell mass formed from the cells at the opening 3. Can be implemented. For this reason, the opening 3 can also be called a culture chamber for culturing the cell mass. The cells and the cell mass will be described later. In addition, although the manufacturing container 10 of this embodiment contains the edge 5, the edge 5 is arbitrary structures and may or may not exist.
　製造容器１０の形成材料は、特に制限されず、例えば、ポリ乳酸、ポリグリコール酸、スチレン樹脂、アクリル樹脂、アクリル・スチレン共重合樹脂、ポリカーボネート樹脂、ポリビニルアルコール樹脂、ポリエチレン樹脂、エチレン・ビニルアルコール共重合樹脂、熱可塑性エラストマー、ポリエチレンテレフタレート樹脂、塩化ビニル樹脂、シリコン樹脂等があげられる。製造容器１０は、例えば、１種類の形成材料から形成されてもよいし、２種類以上の形成材料から形成されてもよい。また、後者の場合、製造容器１０の各構成は、例えば、異なる形成材料から構成されてもよい。 The material for forming the production container 10 is not particularly limited. For example, polylactic acid, polyglycolic acid, styrene resin, acrylic resin, acrylic / styrene copolymer resin, polycarbonate resin, polyvinyl alcohol resin, polyethylene resin, ethylene / vinyl alcohol Examples thereof include polymer resins, thermoplastic elastomers, polyethylene terephthalate resins, vinyl chloride resins, and silicon resins. The manufacturing container 10 may be formed from, for example, one type of forming material, or may be formed from two or more types of forming materials. Moreover, in the latter case, each structure of the manufacturing container 10 may be comprised from a different forming material, for example.
　本実施形態において、製造容器１０は、１つの有底筒状の容器１を含むが、本発明はこれに限定されず、複数の有底筒状の容器１を含んでもよい。前記複数は、２以上であればよく、その上限は、特に制限されない。前記複数は、例えば、６、１２、２４、４８、９６、３８４等があげられる。容器１の筒の形状は、特に制限されず、例えば、円筒状、角筒状等があげられる。容器１の容積、内径、および高さ等は、特に制限されない。 In the present embodiment, the production container 10 includes one bottomed cylindrical container 1, but the present invention is not limited to this, and may include a plurality of bottomed cylindrical containers 1. The plurality may be two or more, and the upper limit is not particularly limited. Examples of the plurality include 6, 12, 24, 48, 96, and 384. The shape of the tube of the container 1 is not particularly limited, and examples thereof include a cylindrical shape and a rectangular tube shape. The volume, inner diameter, height, and the like of the container 1 are not particularly limited.
　本実施形態の製造容器１０において、容器１の底部２は、平面状であるが、本発明はこれに限定されず、例えば、曲面状でもよい。また、容器１の底部２は、複数の開口部３を含むが、本発明はこれに限定されず、例えば、１つの開口部３を含んでもよい。底部２が複数の開口部３を有する場合、開口部３の個数は、２以上であればよく、その上限は、特に制限されない。底部２の単位面積当たりの開口部３の個数は、特に制限されず、例えば、１０～１０００個／ｃｍ２、２０～１０００個／ｃｍ２である。 In the manufacturing container 10 of the present embodiment, the bottom 2 of the container 1 is planar, but the present invention is not limited to this, and may be curved, for example. Moreover, although the bottom part 2 of the container 1 contains the some opening part 3, this invention is not limited to this, For example, you may include the one opening part 3. FIG. When the bottom part 2 has the some opening part 3, the number of the opening parts 3 should just be two or more, and the upper limit in particular is not restrict | limited. The number of openings 3 per unit area of the bottom 2 is not particularly limited, and is, for example, 10 to 1000 pieces / cm 2 and 20 to 1000 pieces / cm 2 .
　本実施形態の製造容器１０において、開口部３は、底部２の内面側から外面側に向かってテーパー状であるが、本発明はこれに限定されず、底部２の外面側における開口部３の径の大きさが、インクを吐出して立体造形を行う立体造形装置のインク吐出口の径の大きさ以下である任意の形状としてよい。前記インクを吐出して立体造形を行う立体造形装置は、例えば、インクジェット式の立体造形装置があげられる。前記インク吐出口は、例えば、前記立体造形装置において、インク収容部に収容されたインクを吐出するインクジェットヘッドの開口部があげられる。具体的に、開口部３の径（内径）は、例えば、５０～１０００μｍ、２００～７００μｍ、３００～５００μｍである。開口部３の径をこのような長さとすることで、例えば、約２００μｍの細胞塊を作製できる。開口部３の径は、特に制限されず、例えば、製造した細胞塊の大きさに応じて、適宜設定できる。開口部３の径は、例えば、底部２の内側面から外側面に向かって同一の径であってもよいし、異なる径であってもよいが、好ましくは、後者である。また、後者の場合、開口部３の径は、例えば、開口部３への細胞の導入が容易になることから、底部２の内側面から外側面に向かって、減少することが好ましい。開口部３の高さは、特に制限されず、例えば、作製する細胞塊の有する所望の径に応じて適宜設定できる。開口部３の高さは、例えば、前記所望の径を有する細胞塊の半径Ｒに対して１～１０倍、１．５～５倍の高さである。前記細胞塊の半径Ｒは、例えば、短径でもよいし、長径でもよいし、平均径でもよい（以下、同様）。 In the manufacturing container 10 of the present embodiment, the opening 3 is tapered from the inner surface side to the outer surface side of the bottom portion 2, but the present invention is not limited to this, and the opening portion 3 on the outer surface side of the bottom portion 2. The diameter may be an arbitrary shape that is equal to or smaller than the diameter of the ink discharge port of the three-dimensional modeling apparatus that performs three-dimensional modeling by discharging ink. Examples of the three-dimensional modeling apparatus that performs three-dimensional modeling by discharging the ink include an ink jet type three-dimensional modeling apparatus. Examples of the ink discharge port include an opening of an ink jet head that discharges ink stored in an ink storage unit in the three-dimensional modeling apparatus. Specifically, the diameter (inner diameter) of the opening 3 is, for example, 50 to 1000 μm, 200 to 700 μm, or 300 to 500 μm. By setting the diameter of the opening 3 to such a length, for example, a cell mass of about 200 μm can be produced. The diameter of the opening 3 is not particularly limited, and can be appropriately set according to the size of the produced cell mass, for example. The diameter of the opening 3 may be, for example, the same diameter from the inner side surface of the bottom part 2 toward the outer side surface or may be a different diameter, but is preferably the latter. In the latter case, the diameter of the opening 3 is preferably reduced from the inner surface to the outer surface of the bottom 2 because, for example, cells can be easily introduced into the opening 3. The height in particular of the opening part 3 is not restrict | limited, For example, it can set suitably according to the desired diameter which the cell mass to produce has. The height of the opening 3 is, for example, 1 to 10 times and 1.5 to 5 times the radius R of the cell mass having the desired diameter. The radius R of the cell mass may be, for example, a short diameter, a long diameter, or an average diameter (hereinafter the same).
　本実施形態の製造容器１０において、縁５は、側壁４から外側に突出しているが、本発明はこれに限定されず、任意の形状とできる。製造容器１０を後述する細胞インクの製造キットの回収容器に配置した際に、縁５が、前記回収容器の内部に製造容器１０を保持し、前記回収容器の底部の内面と製造容器１０の底部の外面とを、一定距離を保つように、製造容器１０に配置されている場合、縁５は、例えば、保持部ともいうことができる。前記一定距離については、後述する。 In the manufacturing container 10 of the present embodiment, the edge 5 protrudes outward from the side wall 4, but the present invention is not limited to this, and can have any shape. When the production container 10 is arranged in a collection container of a cell ink production kit, which will be described later, the edge 5 holds the production container 10 inside the collection container, and the inner surface of the bottom part of the collection container and the bottom part of the production container 10 When the outer surface is arranged in the production container 10 so as to maintain a certain distance, the edge 5 can also be referred to as a holding portion, for example. The fixed distance will be described later.
＜細胞インクの製造キット＞
　本発明の細胞インクの製造キット（以下、「製造キット」ともいう。）は、前述のように、前記本発明の細胞インクの製造容器と、細胞塊を回収するための回収容器とを含み、前記回収容器は、有底筒状であり、内部に前記細胞インクの製造容器を収容可能であることを特徴とする。本発明の製造キットは、前記本発明の製造容器を含むことが特徴であり、その他の構成および条件は、特に制限されない。本発明の製造キットによれば、例えば、後述する立体造形物の造形が可能な細胞インクを製造可能である。本発明の製造キットは、例えば、前記本発明の細胞インク等の説明を援用できる。 <Cell ink manufacturing kit>
The cell ink manufacturing kit of the present invention (hereinafter also referred to as “manufacturing kit”) includes the cell ink manufacturing container of the present invention and a recovery container for recovering the cell mass, as described above. The collection container has a bottomed cylindrical shape, and can accommodate the cell ink production container therein. The production kit of the present invention is characterized by including the production container of the present invention, and other configurations and conditions are not particularly limited. According to the production kit of the present invention, for example, a cell ink capable of modeling a three-dimensional model to be described later can be manufactured. For example, the description of the cell ink of the present invention can be used for the production kit of the present invention.
　本発明の製造キットの一例について、図面を参照し説明する。また、図面においては、説明の便宜上、各部の構造は適宜簡略化して示す場合があり、各部の寸法比等は、実際とは異なり、模式的に示す場合がある。 An example of the production kit of the present invention will be described with reference to the drawings. In the drawings, for convenience of explanation, the structure of each part may be simplified as appropriate, and the dimensional ratio of each part may be schematically shown, unlike the actual case.
　図２（Ａ）は、本実施形態の製造キットの模式分解斜視図であり、（Ｂ）は、前記製造キットにおいて、前記製造容器を前記回収容器に収容した状態の模式透視斜視図であり、（Ｃ）は、（Ｂ）において、II－II方向からみた模式断面図である。図２（Ａ）～（Ｃ）に示すように、本実施形態の製造キット２０は、前記実施形態の製造容器１０および回収容器１１を含む。製造容器１０は、前述の通りである。回収容器１１は、有底筒状であり、上端側に保持部６を含む。製造キット２０において、細胞インクの製造は、例えば、製造容器１０の有底筒状の容器１の内側に培養液および細胞を導入し、開口部３で細胞および前記細胞から形成された細胞塊を培養し、前記培養後の細胞塊を開口部３の内面側から外面側に導出し、回収容器１１で回収することにより実施できる。なお、本実施形態の製造キット２０は、縁５および保持部６を含むが、縁５および保持部６は、任意の構成であり、あってもよいし、なくてもよいし、いずれか１つを含んでもよいし、両者を含んでもよい。 FIG. 2A is a schematic exploded perspective view of the production kit of the present embodiment, and FIG. 2B is a schematic perspective view of the production kit in which the production container is accommodated in the collection container. (C) is a schematic cross-sectional view taken in the direction II-II in (B). As shown in FIGS. 2A to 2C, the manufacturing kit 20 of the present embodiment includes the manufacturing container 10 and the recovery container 11 of the embodiment. The production container 10 is as described above. The collection container 11 has a bottomed cylindrical shape and includes a holding portion 6 on the upper end side. In the production kit 20, the cell ink is produced, for example, by introducing a culture solution and cells into the inside of the bottomed cylindrical container 1 of the production container 10, and removing the cells and the cell mass formed from the cells at the opening 3. It can be carried out by culturing, extracting the cultured cell mass from the inner surface side to the outer surface side of the opening 3 and collecting it in the collection container 11. In addition, although the manufacturing kit 20 of this embodiment includes the edge 5 and the holding part 6, the edge 5 and the holding part 6 are arbitrary structures, and may or may not be any one. May be included, or both may be included.
　回収容器１１の形成材料は、特に制限されず、例えば、「製造容器１０」を「回収容器１１」に読み替えて、製造容器１０の形成材料の説明を援用できる。回収容器１１の形成材料は、例えば、製造容器１０と同じ形成材料でもよいし、異なる形成材料でもよい。 The formation material of the collection container 11 is not particularly limited. For example, the description of the formation material of the production container 10 can be cited by replacing “production container 10” with “recovery container 11”. The forming material of the collection container 11 may be, for example, the same forming material as the manufacturing container 10 or a different forming material.
　本実施形態において、回収容器１１は、１つであるが、本発明はこれに限定されず、複数であってもよい。回収容器１１は、有底筒状であればよく、その形状は、特に制限されない。回収容器１１の筒の形状は、例えば、円筒状、角筒状等があげられる。回収容器１１は、例えば、製造容器１０と同じ形状でもよいし、異なる形状でもよい。回収容器１１の数は、例えば、製造容器１１の容器１の数と同じでもよいし、異なってもよいが、好ましくは、前者である。回収容器１１は、内部に製造容器１０を収容可能であればよく、その大きさは、特に制限されない。回収容器１１の容積、内径、および高さ等は、例えば、製造容器１０および製造容器１０内で培養可能な培養液の容積等に応じて、適宜設定できる。 In the present embodiment, the number of the collection containers 11 is one, but the present invention is not limited to this, and may be plural. The collection container 11 may be a bottomed cylindrical shape, and the shape is not particularly limited. Examples of the shape of the tube of the collection container 11 include a cylindrical shape and a rectangular tube shape. For example, the collection container 11 may have the same shape as the production container 10 or a different shape. The number of collection containers 11 may be, for example, the same as or different from the number of containers 1 of the production container 11, but is preferably the former. The collection container 11 is not particularly limited as long as the production container 10 can be accommodated therein, and its size is not particularly limited. The volume, inner diameter, height, and the like of the collection container 11 can be appropriately set according to, for example, the production container 10 and the volume of the culture solution that can be cultured in the production container 10.
　回収容器１１は、例えば、市販のディッシュ、市販のマルチウェルプレート等でもよい。前記ディッシュは、例えば、φ３ｃｍ、φ５ｃｍ、φ１０ｃｍのディッシュ等があげられる。前記マルチウェルプレートは、例えば、６、１２、２４、４８、９６、３８４ウェルのプレート等があげられる。この場合、本発明の製造キットは、例えば、回収容器１１に、製造容器１０を配置することにより使用できる。 The collection container 11 may be, for example, a commercially available dish or a commercially available multiwell plate. Examples of the dish include φ3 cm, φ5 cm, and φ10 cm dishes. Examples of the multi-well plate include 6, 12, 24, 48, 96, and 384 well plates. In this case, the production kit of the present invention can be used, for example, by arranging the production container 10 in the collection container 11.
　本実施形態において、保持部６は、回収容器１１の上側端として形成されているが、本発明はこれに限定されず、製造容器１０を回収容器１１の内部に保持し、且つ回収容器１１の底部の内面と、製造容器１０の底部２の外面とを、一定距離を保つように配置できる任意の形状としてよい。保持部６が、製造容器１０を保持する方法は、特に制限されず、例えば、回収容器１１が、保持部６を介して製造容器１０を保持してもよいし、回収容器１１が、保持部６を介して製造容器１０に保持されていてもよい。前記一定距離は、特に制限されず、例えば、０ｍｍより長い。 In the present embodiment, the holding unit 6 is formed as the upper end of the recovery container 11, but the present invention is not limited to this, and the manufacturing container 10 is held inside the recovery container 11 and the recovery container 11 The inner surface of the bottom portion and the outer surface of the bottom portion 2 of the manufacturing container 10 may have any shape that can be arranged so as to maintain a certain distance. The method by which the holding unit 6 holds the production container 10 is not particularly limited. For example, the collection container 11 may hold the production container 10 via the holding unit 6, or the collection container 11 may be the holding unit. 6 may be held in the production container 10 via The fixed distance is not particularly limited, and is longer than 0 mm, for example.
　本実施形態において、回収容器１１は、有底筒状の容器であるが、本発明はこれに限定されず、回収容器１１は、例えば、インクを吐出して立体造形を行なう立体造形装置用のインクタンクであってもよい。この場合、回収容器１１は、さらに、インク吐出部を含む。回収容器１１において、前記インク吐出部は、インクを吐出できるように配置されればよく、その位置は、特に制限されない。回収容器１１における前記インク吐出部の位置は、例えば、回収容器１１の底面、側面等があげられる。 In the present embodiment, the collection container 11 is a bottomed cylindrical container, but the present invention is not limited to this, and the collection container 11 is, for example, for a three-dimensional modeling apparatus that performs three-dimensional modeling by discharging ink. It may be an ink tank. In this case, the collection container 11 further includes an ink ejection unit. In the collection container 11, the ink ejecting portion may be disposed so as to eject ink, and the position thereof is not particularly limited. Examples of the position of the ink ejection unit in the collection container 11 include the bottom surface and the side surface of the collection container 11.
　本発明の製造キットは、さらに、製造容器１０または回収容器１１の開口を塞ぐ蓋を含んでもよい。本発明の製造キットは、前記蓋を含むことで、例えば、外部からの異物の混入を防止できる。また、回収容器１１がインクジェット式３Ｄプリンターのインクタンクである場合、前記蓋を前記インクタンクに装着することで、例えば、後述するインクジェット式３Ｄプリンターのインクカートリッジ（以下、「インクカートリッジ」ともいう。）として使用できる。前記蓋は、例えば、製造容器１０または回収容器１１を覆う蓋でもよいし、製造容器１０または回収容器１１に内接する蓋でもよい。 The production kit of the present invention may further include a lid that closes the opening of the production container 10 or the collection container 11. By including the lid, the production kit of the present invention can prevent foreign matters from being mixed in, for example. Further, when the collection container 11 is an ink tank of an ink jet 3D printer, for example, an ink cartridge (hereinafter also referred to as “ink cartridge”) of an ink jet 3D printer described later is mounted on the ink tank. ) Can be used. The lid may be, for example, a lid that covers the production container 10 or the collection container 11, or a lid that is inscribed in the production container 10 or the collection container 11.
＜細胞インクの製造方法＞
　本発明の細胞インクの製造方法は、前述のように、前記本発明の細胞インクの製造容器を用いて細胞を培養する培養工程、および培養された細胞塊を回収する回収工程を含み、前記培養工程において、培養液の存在下、前記細胞インクの製造容器の前記開口部において、細胞を培養し、前記回収工程において、前記開口部から、前記細胞インクの製造容器の外部に導出された細胞塊を回収することを特徴とする。本発明の細胞インクの製造方法によれば、例えば、後述する立体造形装置により立体を造形した際のインク吐出口の目詰まりが低減された細胞インクを製造できる。本発明の細胞インクの製造方法は、例えば、前記本発明の細胞インク等の説明を援用できる。 <Method for producing cell ink>
As described above, the method for producing a cell ink of the present invention includes a culture step of culturing cells using the cell ink production container of the present invention, and a recovery step of recovering the cultured cell mass. In the step, cells are cultured in the opening of the cell ink production container in the presence of a culture solution, and in the collecting step, the cell mass led out of the cell ink production container from the opening. It is characterized by collect | recovering. According to the method for producing cell ink of the present invention, for example, cell ink in which clogging of ink discharge ports when a solid is modeled by a three-dimensional modeling apparatus described later can be manufactured. For example, the description of the cell ink of the present invention can be used in the method for producing the cell ink of the present invention.
　前記細胞の種類は、特に制限されず、任意の細胞とできる。前記細胞は、例えば、培養後に得られる細胞塊を形成する細胞（以下、「目的細胞」ともいう。）と同じ細胞でもよいし、異なる細胞でもよい。前記目的細胞は、例えば、前記細胞インクを使用し造形する立体造形物の種類に応じて適宜決定できる。具体的に、前記立体造形物が肝臓である場合、前記目的細胞は、例えば、肝細胞等の実質細胞、類洞壁細胞、筋線維芽細胞、胆管上皮細胞、結合組織細胞等の非実質細胞等があげられる。前記目的細胞が前記細胞と異なる場合、前記細胞は、例えば、前記目的細胞に分化する細胞等があげられる。前記目的細胞に分化する細胞は、特に制限されず、例えば、ｉＰＳ細胞、ＥＳ細胞等の多能性幹細胞、体性幹細胞等の幹細胞、前駆細胞等があげられる。 The type of the cell is not particularly limited and can be any cell. The cell may be, for example, the same cell as a cell forming a cell mass obtained after culture (hereinafter also referred to as “target cell”) or a different cell. The target cell can be appropriately determined according to, for example, the type of three-dimensional structure to be formed using the cell ink. Specifically, when the three-dimensional model is a liver, the target cells are, for example, parenchymal cells such as hepatocytes, sinusoidal wall cells, myofibroblasts, bile duct epithelial cells, connective tissue cells and the like. Etc. When the target cell is different from the cell, examples of the cell include a cell that differentiates into the target cell. The cells that differentiate into the target cells are not particularly limited, and examples thereof include pluripotent stem cells such as iPS cells and ES cells, stem cells such as somatic stem cells, and progenitor cells.
　前記細胞の由来は、特に制限されず、例えば、ヒト、非ヒト哺乳類、鳥類、は虫類、魚介類等があげられる。 The origin of the cells is not particularly limited, and examples thereof include humans, non-human mammals, birds, reptiles, and seafood.
　前記培養液は、特に制限されず、培養する前記細胞の種類に応じて、適宜決定できる。前記培養液は、さらに、その他の成分を含んでもよい。前記その他の成分は、例えば、前記細胞を増殖させる増殖因子、前記細胞を分化させる分化因子等があげられる。前記増殖因子および前記分化因子は、特に制限されず、前記細胞の種類に応じて適宜決定できる。 The culture medium is not particularly limited, and can be appropriately determined according to the type of the cell to be cultured. The culture solution may further contain other components. Examples of the other components include a growth factor for growing the cell, a differentiation factor for differentiating the cell, and the like. The growth factor and the differentiation factor are not particularly limited, and can be appropriately determined according to the type of the cell.
　前記細胞塊を構成する細胞の種類は、特に制限されず、前記細胞および培養等の条件により適宜調整できる。 The type of cells constituting the cell mass is not particularly limited and can be appropriately adjusted depending on the conditions of the cells and culture.
　前記培養工程は、前記本発明の細胞インクの製造容器を用い、前記細胞インクの製造容器の前記開口部において、培養液の存在下、細胞を培養する。具体的に、前記培養工程は、例えば、前記製造容器の有底筒状の容器内に、前記製造容器の上側の開口から前記細胞および前記培養液を導入し、培養する。前記細胞は、例えば、前記培養開始時において、分散状態でもよいし、細胞塊の状態でもよいが、好ましくは、前者である。また、導入する前記細胞の種類は、例えば、１種類でもよいし、２種類以上でもよい。前記培養工程において、前記細胞は、例えば、前記製造容器の前記開口部において培養してもよいし、前記製造容器の有底筒状の容器内において培養してもよいし、前記製造容器の前記開口部から形成された懸滴内において培養してもよいし、これらの組合せで培養してもよいが、例えば、ランダムな細胞同士の接触による不均一な細胞塊の発生を抑制でき、より均一な細胞塊を作製できることから、前記開口部において培養することが好ましい。前記開口部において培養する場合、前記細胞の導入後、例えば、前記製造容器を遠心等し、前記細胞を前記開口部に導入してもよい。 In the culturing step, the cell ink production container of the present invention is used, and the cells are cultured in the presence of a culture solution in the opening of the cell ink production container. Specifically, in the culture step, for example, the cells and the culture solution are introduced into the bottomed cylindrical container of the production container from the upper opening of the production container and cultured. For example, the cells may be in a dispersed state or a cell mass at the start of the culture, but the former is preferable. Moreover, the type of the cells to be introduced may be, for example, one type or two or more types. In the culturing step, the cells may be cultured, for example, in the opening of the manufacturing container, in a bottomed cylindrical container of the manufacturing container, or in the manufacturing container. The cells may be cultured in a hanging drop formed from the opening, or may be cultured in combination thereof. For example, the generation of a non-uniform cell mass due to contact between random cells can be suppressed, and more uniform. It is preferable to culture in the opening because a cell mass can be produced. When culturing in the opening, after the introduction of the cell, for example, the production container may be centrifuged to introduce the cell into the opening.
　前記培養における培養条件は、特に制限されず、前記細胞の種類に応じて、適宜設定できる。前記培養工程において、前記培養温度は、例えば、３６～３７℃であり、Ｏ２分圧は、例えば、１～２１％であり、ＣＯ２分圧は、例えば、５～６％である。 The culture conditions in the culture are not particularly limited and can be appropriately set according to the type of the cell. In the culture step, the culture temperature is, for example, 36 to 37 ° C., the O 2 partial pressure is, for example, 1 to 21%, and the CO 2 partial pressure is, for example, 5 to 6%.
　前記培養における培養期間は、特に制限されず、例えば、細胞塊を形成する期間があげられ、前記細胞の種類に応じて、適宜設定できる。前記培養期間は、例えば、１２時間～３０日である。具体的に、前記細胞が肝実質細胞の場合、前記培養期間は、例えば、２４時間～５日である。前記培養期間は、例えば、本発明の製造方法により得られた細胞インクを用いて立体造形物の造形後、前記細胞塊間の接着が促進され、前記立体造形物を構成する細胞の細胞死を抑制できることから、前記細胞塊が、前記細胞塊から分泌された細胞外マトリックスを前記細胞塊の周囲に保持するまでの期間であることが好ましい。 The culture period in the culture is not particularly limited, and examples thereof include a period for forming a cell mass, and can be appropriately set according to the type of the cell. The culture period is, for example, 12 hours to 30 days. Specifically, when the cells are hepatocytes, the culture period is, for example, 24 hours to 5 days. In the culture period, for example, after the formation of the three-dimensional structure using the cell ink obtained by the production method of the present invention, adhesion between the cell masses is promoted, and cell death of the cells constituting the three-dimensional structure is reduced. Since it can suppress, it is preferable that it is the period until the said cell mass hold | maintains the extracellular matrix secreted from the said cell mass around the said cell mass.
　前記培養工程において、前記製造容器を含む製造キットを使用する場合、前記製造容器を前記回収容器の内部に収容し、前記培養液の存在下、細胞を培養することが好ましい。この場合、前記培養液は、例えば、前記製造容器に加え、前記回収容器に導入してもよい。 In the culturing step, when a production kit including the production container is used, it is preferable that the production container is accommodated in the collection container and the cells are cultured in the presence of the culture solution. In this case, for example, the culture solution may be introduced into the collection container in addition to the production container.
　本発明の細胞インクの製造方法は、例えば、前記回収工程において回収した細胞塊を、そのまま細胞インクとして使用できることから、前記回収工程に先立ち、前記培養液を前記細胞インク用溶液と交換する溶液交換工程を含むことが好ましい。前記細胞インク用溶液は、例えば、前記細胞インクにおける前記細胞塊以外の成分を含む溶液等があげられる。前記細胞塊以外の成分は、特に制限されず、例えば、脂質、前記細胞外マトリックス、生体材料、刺激応答性分子等があげられる。前記刺激応答性分子は、例えば、外部刺激により物性が変化する物質である。前記外部刺激は、例えば、機械的刺激でもよいし、電磁的刺激でもよいし、化学的刺激でもよい。前記機械的刺激は、例えば、物理的刺激があげられ、具体的には、圧力、音等の刺激があげられる。前記電磁的刺激は、例えば、光、熱等の刺激があげられる。前記物性の変化は、例えば、固形化、ゲル化、液化等があげられる。前記細胞インク用溶液は、例えば、１種類の前記細胞塊以外の成分を含んでもよいし、２種類以上の前記細胞塊以外の成分を含んでもよい。前記細胞インク用溶液を構成する溶液は、特に制限されず、例えば、リン酸緩衝液等があげられる。 In the method for producing a cell ink of the present invention, for example, since the cell mass collected in the collecting step can be used as a cell ink as it is, solution exchange for exchanging the culture solution with the cell ink solution prior to the collecting step. It is preferable to include a process. Examples of the cell ink solution include a solution containing a component other than the cell mass in the cell ink. Components other than the cell mass are not particularly limited, and examples thereof include lipids, the extracellular matrix, biomaterials, and stimulus-responsive molecules. The stimulus-responsive molecule is a substance whose physical properties are changed by an external stimulus, for example. The external stimulus may be, for example, a mechanical stimulus, an electromagnetic stimulus, or a chemical stimulus. Examples of the mechanical stimulus include a physical stimulus, and specifically, a stimulus such as pressure and sound. Examples of the electromagnetic stimulation include stimulation such as light and heat. Examples of the change in physical properties include solidification, gelation, and liquefaction. The cell ink solution may include, for example, a component other than one type of the cell mass, or may include two or more types of components other than the cell mass. The solution constituting the cell ink solution is not particularly limited, and examples thereof include a phosphate buffer solution.
　前記液体交換工程において、前記培養液と前記細胞インク用溶液との交換は、例えば、前記製造容器内の培養液を除去し、前記細胞インク用溶液を前記製造容器内に導入することにより実施できる。前記培養液の除去は、例えば、前記製造容器の有底筒状の容器内の培養液の除去でもよいし、前記開口部の培養液の除去でもよいし、両者の除去でもよい。前記除去および導入は、例えば、前記製造容器の上側の開口から実施する。 In the liquid exchange step, the culture medium and the cell ink solution can be exchanged by, for example, removing the culture liquid in the production container and introducing the cell ink solution into the production container. . The removal of the culture solution may be, for example, the removal of the culture solution in the bottomed cylindrical container of the production vessel, the removal of the culture solution in the opening, or the removal of both. The removal and introduction are performed, for example, from the upper opening of the production container.
　前記回収工程は、前記開口部から、前記細胞インクの製造容器の外部に導出された細胞塊を回収する。前記細胞塊を前記開口部から前記製造容器の外部へ導出する方法は、特に制限されない。前記導出は、例えば、前記製造容器の内部から前記製造容器の外部に前記細胞塊を含む培養液を自然滴下させることにより実施してもよいし、前記製造容器の内部から外部方向に圧力をかけることにより、前記細胞塊を含む培養液を前記開口部から押出することにより実施してもよいし、前記製造容器の内部から外部方向に、前記細胞塊を含む培養液を吸出すことにより実施してもよい。 The collection step collects the cell mass led out of the cell ink production container from the opening. The method for deriving the cell mass from the opening to the outside of the production container is not particularly limited. The derivation may be performed, for example, by spontaneously dropping a culture solution containing the cell mass from the inside of the production container to the outside of the production container, or applying pressure from the inside of the production container to the outside. Thus, it may be carried out by extruding the culture solution containing the cell mass from the opening, or by sucking out the culture solution containing the cell mass from the inside to the outside of the production container. May be.
　また、前記回収工程において、前記製造容器を含む製造キットを使用する場合、前記開口部から、前記細胞インクの製造容器の外部の前記回収容器に導出された細胞塊を回収することが好ましい。また、前記回収工程は、例えば、前記細胞塊に対するダメージを抑制し、前記細胞塊を回収できることから、前記回収容器に回収溶液を導入し、前記開口部と前記回収溶液とを接触させることにより、前記開口部から、前記細胞インクの製造容器の外部の前記回収容器に導出された細胞塊を回収することがより好ましい。前記回収溶液は、例えば、前記細胞インク用溶液等があげられる。前記細胞塊の回収は、例えば、前記製造容器を前記回収容器の内部に収容し、前記細胞塊を前記開口部から前記製造容器の外部へ導出することにより実施できる。前記製造容器を含む製造キットを使用する場合、前記回収工程において、前記回収容器から前記細胞インクの製造容器を取り外し、前記回収容器内の前記細胞塊を回収することが好ましい。 In the recovery step, when a manufacturing kit including the manufacturing container is used, it is preferable to recover the cell mass derived from the opening to the recovery container outside the cell ink manufacturing container. In addition, the recovery step, for example, suppresses damage to the cell mass and can collect the cell mass, so that by introducing a recovery solution into the recovery container and bringing the opening and the recovery solution into contact with each other, More preferably, the cell mass led out to the collection container outside the cell ink production container is collected from the opening. Examples of the recovered solution include the cell ink solution. The collection of the cell mass can be carried out, for example, by housing the production container inside the collection container and leading the cell mass out of the production container through the opening. When a production kit including the production container is used, it is preferable that in the collection step, the cell ink production container is removed from the collection container and the cell mass in the collection container is collected.
　このようにして、前記回収工程で回収した細胞塊を細胞インクとしてもよいし、前記回収した細胞塊を、さらに、前記細胞インク用溶液に分散した溶液を細胞インクとしてもよい。前記細胞インクは、例えば、１種類の細胞塊を含んでもよいし、２種類以上の細胞塊を含んでもよい。 Thus, the cell mass recovered in the recovery step may be used as cell ink, or a solution obtained by dispersing the recovered cell mass in the cell ink solution may be used as cell ink. The cell ink may include, for example, one type of cell mass or two or more types of cell mass.
＜細胞インク＞
　本発明の細胞インクは、前述のように、前記本発明の細胞インクの製造方法により得られることを特徴とする。本発明の細胞インクは、前記本発明の細胞インクの製造方法により得られたことが特徴であり、その他の構成および条件は、特に制限されない。本発明の細胞インクによれば、例えば、後述するインクジェット式３Ｄプリンターにより立体造形物を造形した際に、前記インクジェット式３Ｄプリンターのインク吐出口の目詰まりを低減できる。また、本発明の細胞インクによれば、例えば、前述の細胞インクの効果を得ることができる。本発明の細胞インクは、例えば、前記本発明の細胞インク等の説明を援用できる。 <Cellular ink>
As described above, the cell ink of the present invention is obtained by the method for producing the cell ink of the present invention. The cell ink of the present invention is obtained by the method for producing the cell ink of the present invention, and other configurations and conditions are not particularly limited. According to the cell ink of the present invention, for example, when a three-dimensional model is modeled by an ink jet 3D printer described later, clogging of the ink discharge port of the ink jet 3D printer can be reduced. Moreover, according to the cell ink of the present invention, for example, the effect of the cell ink described above can be obtained. For example, the description of the cell ink of the present invention can be used for the cell ink of the present invention.
＜インクカートリッジ＞
　本発明のインクを吐出して立体造形を行う立体造形装置用のインクカートリッジは、前述のように、前記本発明の細胞インクを含むことを特徴とする。本発明のインクカートリッジは、前記本発明の細胞インクを含むことが特徴であり、その他の構成および条件は、特に制限されない。本発明のインクカートリッジによれば、例えば、インクジェット式３Ｄプリンターを用いて、簡便に立体造形物を造形できる。本発明のインクカートリッジは、例えば、前記本発明の細胞インク等の説明を援用できる。 <Ink cartridge>
As described above, an ink cartridge for a three-dimensional modeling apparatus that performs three-dimensional modeling by discharging the ink of the present invention includes the cell ink of the present invention. The ink cartridge of the present invention is characterized by containing the cell ink of the present invention, and other configurations and conditions are not particularly limited. According to the ink cartridge of the present invention, it is possible to easily model a three-dimensional model using, for example, an ink jet 3D printer. For the ink cartridge of the present invention, for example, the description of the cell ink of the present invention can be cited.
　本発明のインクカートリッジは、例えば、１種類の細胞インクを含んでもよいし、２種類以上の細胞インクを含んでもよい。後者の場合、前記インクカートリッジは、例えば、前記細胞インクを同一の容器に混合、または未混同で含んでもよいし、別個の容器に含んでもよい。前記２種類以上の細胞インクを別個の容器に含む場合、本発明のインクカートリッジは、例えば、インクセットということもできる。 The ink cartridge of the present invention may contain, for example, one type of cell ink or two or more types of cell ink. In the latter case, the ink cartridge may contain, for example, the cell ink in the same container or not mixed, or may be contained in a separate container. When the two or more types of cell ink are contained in separate containers, the ink cartridge of the present invention can also be referred to as an ink set, for example.
＜立体造形物の製造方法＞
　本発明の立体造形物の製造方法は、前述のように、細胞インクを用いる立体造形工程を含み、前記立体造形工程が、造形対象物の立体データに基づき、インクを吐出して立体造形を行う立体造形装置のインク吐出口から、前記細胞インクを吐出し、立体造形物を造形し、前記細胞インクが、前記本発明の細胞インク、および前記本発明の細胞インクの製造方法により得られた細胞インクの少なくとも一方であることを特徴とする。本発明の立体造形物の製造方法は、前記本発明の細胞インク、および前記本発明の細胞インクの製造方法により得られた細胞インクの少なくとも一方を用い、立体造形物を造形することが特徴であり、その他の工程および条件は、特に制限されない。本発明の立体造形物の製造方法によれば、例えば、食品、移植用臓器等の細胞から形成される立体造形物が製造可能である。また、本発明の立体造形物の製造方法は、例えば、血管等を造形することで、内部への栄養、酸素等の供給を行うことができる立体造形物も造形可能である。このため、本発明の立体造形物の製造方法によれば、例えば、培養により臓器等を造形した場合と比較して、生体の臓器等により近い大きさの臓器等が造形できる。本発明の立体造形物の製造方法は、例えば、前記本発明の細胞インク等の説明を援用できる。また、本発明の立体造形物の製造方法に用いる細胞インクは、前記細胞塊を含むため、例えば、同じ大きさの立体造形物を造形する場合、単細胞を含む細胞インクと比較して、より短時間で造形できる。また、本発明の立体造形物の製造方法によれば、例えば、前記細胞インクを用いることにより、より短時間で立体造形物を造形できることから、造形した前記立体造形物を構成する細胞の細胞死を抑制できる。さらに、前記細胞インクは、例えば、前記細胞塊を構成する細胞の細胞表面における細胞接着分子の密度が損なわれていない。このため、本発明の立体造形物の製造方法によれば、例えば、前記細胞インクを用いて前記立体造形物として食品を造形した場合、鮮度が高く、本来の食味に近い食品が造形できる。また、本発明の立体造形物の製造方法によれば、例えば、前記細胞インクを用いて前記立体造形物として臓器を造形した場合、前記臓器の機能が維持された臓器が造形できる。 <Method for manufacturing a three-dimensional model>
As described above, the method for manufacturing a three-dimensional object of the present invention includes a three-dimensional object forming process using cell ink, and the three-dimensional object forming process performs three-dimensional object formation by discharging ink based on the three-dimensional data of the object to be formed. The cell ink is ejected from the ink discharge port of the three-dimensional modeling apparatus to form a three-dimensional modeled object, and the cell ink is obtained by the cell ink of the present invention and the method for producing the cell ink of the present invention. It is at least one of ink. The method for producing a three-dimensional structure of the present invention is characterized in that a three-dimensional structure is formed using at least one of the cell ink of the present invention and the cell ink obtained by the method of manufacturing the cell ink of the present invention. The other steps and conditions are not particularly limited. According to the method for producing a three-dimensional structure of the present invention, for example, a three-dimensional structure formed from cells such as foods and transplanted organs can be produced. Moreover, the manufacturing method of the three-dimensional molded item of this invention can also model the three-dimensional molded item which can supply a nutrient, oxygen, etc. to an inside by modeling a blood vessel etc., for example. For this reason, according to the manufacturing method of the three-dimensional molded item of this invention, compared with the case where an organ etc. are modeled by culture | cultivation, for example, the organ of a magnitude | size close | similar to a biological organ etc. can be modeled. For example, the description of the cell ink of the present invention can be used in the method for producing a three-dimensional structure of the present invention. In addition, since the cell ink used in the method for producing a three-dimensional structure of the present invention includes the cell mass, for example, when forming a three-dimensional structure of the same size, the cell ink is shorter than a cell ink including a single cell. Can be shaped in time. Moreover, according to the manufacturing method of the three-dimensional modeled object of the present invention, for example, by using the cell ink, the three-dimensional modeled object can be modeled in a shorter time. Therefore, the cell death of the cells constituting the modeled three-dimensional modeled object Can be suppressed. Furthermore, in the cell ink, for example, the density of cell adhesion molecules on the cell surface of the cells constituting the cell mass is not impaired. For this reason, according to the manufacturing method of the three-dimensional molded item of this invention, when food is modeled as the said three-dimensional molded item using the said cell ink, the food with high freshness and the original taste can be modeled. Further, according to the method for manufacturing a three-dimensional structure of the present invention, for example, when an organ is formed as the three-dimensional structure using the cell ink, an organ in which the function of the organ is maintained can be formed.
　前記立体データは、例えば、目的の立体造形物の三次元的な構造を規定するデータ等があげられる。前記立体データは、例えば、三次元ＣＴ（computed tomography）画像、三次元ＭＲＩ（magnetic resonance imaging）画像、三次元ＰＥＴ（positron emission tomography）画像、三次元ＰＥＴ－ＣＴ画像等があげられる。 Examples of the three-dimensional data include data that defines the three-dimensional structure of the target three-dimensional object. Examples of the three-dimensional data include three-dimensional CT (computed tomography) images, three-dimensional MRI (magnetic resonance tomography) images, three-dimensional PET (positron tomography) tomography images, and three-dimensional PET-CT images.
　前記立体データは、例えば、生体の臓器等により近い構造を有する臓器を造形できることから、前記造形対象物を構成する組織の情報を含むことが好ましい。前記情報は、例えば、構造、体積、密度、組織の状態等の情報があげられる。具体的に、前記組織情報は、例えば、上皮組織、結合組織、神経組織、血管組織、脂肪組織、筋肉組織等の組織の情報があげられる。前記組織情報は、例えば、１種類でもよいし、２種類以上でもよい。 The three-dimensional data preferably includes information on a tissue constituting the modeling target because, for example, an organ having a structure closer to a living organ or the like can be modeled. Examples of the information include information on structure, volume, density, tissue state, and the like. Specifically, examples of the tissue information include information on tissues such as epithelial tissue, connective tissue, nerve tissue, vascular tissue, adipose tissue, and muscle tissue. For example, the organization information may be one type or two or more types.
　前記立体造形物は、特に制限されず、前記細胞から構成される立体造形物であればよい。前記立体造形物は、例えば、食品、臓器等があげられる。前記食品は、特に制限されず、例えば、レバー、トロ、サーロイン、ひれ、ロース等の任意の食品があげられる。前記臓器は、特に制限されず、例えば、肝臓、心臓、腎臓、膵臓、肺等があげられる。前記臓器は、例えば、前記臓器の一部でもよいし、全体でもよい。前記臓器の一部は、例えば、器官等があげられる。 The three-dimensional model is not particularly limited as long as it is a three-dimensional model composed of the cells. Examples of the three-dimensional model include foods and organs. The food is not particularly limited, and examples thereof include any food such as liver, tro, sirloin, fin, and loin. The organ is not particularly limited, and examples thereof include liver, heart, kidney, pancreas, and lung. The organ may be, for example, a part or the whole of the organ. Examples of the organ include an organ.
　使用する前記細胞インクの種類は、特に制限されず、例えば、前記立体造形物に応じて適宜決定できる。使用する前記細胞インクの数は、特に制限されず、例えば、前記造形対象物を構成する細胞の種類数に応じて、適宜設定できる。使用する前記細胞インクの数は、例えば、１種類でもよいし、２種類以上でもよい。後者の場合、各細胞インクが含む細胞塊を構成する細胞の種類は、例えば、同じでもよいし、異なってもよい。また、各細胞インクが含む細胞塊の大きさは、例えば、同じでもよいし、異なってもよい。 The type of the cell ink to be used is not particularly limited and can be appropriately determined according to, for example, the three-dimensional model. The number of the cell inks to be used is not particularly limited, and can be appropriately set according to, for example, the number of types of cells constituting the modeling object. The number of the cell inks used may be, for example, one type or two or more types. In the latter case, the types of cells constituting the cell mass included in each cell ink may be the same or different, for example. Moreover, the size of the cell mass which each cell ink contains may be the same, for example, and may differ.
　本発明の立体造形物の製造方法は、さらに、前記細胞インクを製造する細胞インクの製造工程を含んでもよい。この場合、前記細胞インクの製造工程は、例えば、前記本発明の細胞インクの製造方法により実施できる。また、前記立体造形工程では、例えば、前記製造工程により得られた細胞インクを用いる。 The method for manufacturing a three-dimensional structure of the present invention may further include a cell ink manufacturing process for manufacturing the cell ink. In this case, the manufacturing process of the cell ink can be performed by, for example, the cell ink manufacturing method of the present invention. Moreover, in the said three-dimensional modeling process, the cell ink obtained by the said manufacturing process is used, for example.
　本発明の立体造形物の製造方法は、さらに、前記立体造形工程に先立ち、前記造形対象物の立体データを取得する立体データ取得工程を含んでもよい。前記立体データは、例えば、前記インクジェット式３Ｄプリンターが取得する。前記立体データ取得工程において取得する立体データは、特に制限されず、前記造形対象物に応じて適宜決定できる。具体的に、前記立体造形物が、血管を含む造形対象物である場合、前記立体データ取得工程において、例えば、前記造形対象物の血管組織情報を含む立体データを取得する。 The manufacturing method of the three-dimensional modeled object of the present invention may further include a three-dimensional data acquisition step of acquiring the three-dimensional data of the modeling target prior to the three-dimensional modeling step. The three-dimensional data is acquired by, for example, the inkjet 3D printer. The three-dimensional data acquired in the three-dimensional data acquisition step is not particularly limited, and can be appropriately determined according to the modeling object. Specifically, when the three-dimensional object is a modeling object including a blood vessel, in the three-dimensional data acquisition step, for example, three-dimensional data including blood vessel tissue information of the modeling object is acquired.
　前記立体データの取得方法は、特に制限されず、例えば、前記立体データが記憶された記憶媒体から取得してもよいし、前記造形対象物を測定することにより、前記立体データを取得してもよい。後者の場合、本発明の立体造形物の製造方法は、例えば、前記造形対象物の断層画像を取得する断層画像取得工程と、前記断層画像から前記立体データを作製する立体データ作製工程とを含む。前記断層画像の取得は、例えば、前記造形対象物を、ＣＴ、ＭＲＩ、ＰＥＴ等の断層画像取得装置で測定することにより取得できる。また、前記立体データの作製は、例えば、前記断層画像取得装置で得られた測定データから、前記三次元ＣＴ画像、前記三次元ＭＲＩ画像、前記三次元ＰＥＴ画像、前記三次元ＰＥＴ－ＣＴ画像等の立体データを作製することにより取得できる。具体的に、前記造形対象物がヒトまたは非ヒト動物の臓器であり、前記立体データが血管組織情報を含む立体データである場合、例えば、前記ヒトまたは非ヒト動物に対し、公知の血管造影法を実施することにより、前記造形対象物の断層画像を２枚以上取得する。つぎに、前記２枚以上の断層画像に対し、例えば、公知のボリュームデータ化処理を行なうことで、前記立体データを取得できる。 The method for acquiring the three-dimensional data is not particularly limited. For example, the three-dimensional data may be acquired from a storage medium in which the three-dimensional data is stored, or the three-dimensional data may be acquired by measuring the modeling object. Good. In the latter case, the manufacturing method of the three-dimensional structure of the present invention includes, for example, a tomographic image acquisition step of acquiring a tomographic image of the modeling object and a three-dimensional data generation step of generating the three-dimensional data from the tomographic image. . The acquisition of the tomographic image can be acquired, for example, by measuring the modeling object with a tomographic image acquisition device such as CT, MRI, or PET. The three-dimensional data can be produced, for example, from the measurement data obtained by the tomographic image acquisition apparatus, from the three-dimensional CT image, the three-dimensional MRI image, the three-dimensional PET image, the three-dimensional PET-CT image, etc. Can be obtained by creating the three-dimensional data. Specifically, when the modeling object is an organ of a human or non-human animal and the stereoscopic data is stereoscopic data including vascular tissue information, for example, a known angiography method for the human or non-human animal. As a result, two or more tomographic images of the modeling object are acquired. Next, the three-dimensional data can be acquired by performing, for example, a known volume data conversion process on the two or more tomographic images.
　前記立体造形工程は、前記立体データに基づき、インクを吐出して立体造形を行う立体造形装置のインク吐出口から、前記細胞インクを吐出し、立体造形物を造形する。前記インクジェット式３Ｄプリンターは、例えば、インク収容部とインク吐出手段を含み、前記インク収容部に収容されたインクを前記インク吐出手段によって吐出するインクジェット式３Ｄプリンターである。また、前記インクジェット式３Ｄプリンターは、例えば、前記インク収容部に、前記本発明のインクカートリッジが収容されている。前記立体造形物の造形は、例えば、前記インクジェット式３Ｄプリンターの種類に応じて、適宜実施でき、具体的には、前記インクジェット式３Ｄプリンターに、前記立体データを入力し、デフォルトの設定で造形することにより実施できる。前記インクジェット式３Ｄプリンターは、例えば、１種類のインクを吐出可能なインクジェット式３Ｄプリンターでもよいし、２種類以上のインクを吐出可能なインクジェット式３Ｄプリンターでもよい。前記細胞インクは、例えば、前記本発明の細胞インクに代えて、前記本発明の細胞インクの製造方法により得られた細胞インクを用いてもよい。 In the three-dimensional modeling process, based on the three-dimensional data, the cellular ink is ejected from an ink discharge port of a three-dimensional modeling apparatus that performs ink three-dimensional modeling by discharging ink, thereby modeling a three-dimensional model. The ink jet 3D printer is, for example, an ink jet 3D printer that includes an ink container and an ink discharge unit, and discharges ink stored in the ink container by the ink discharge unit. In the ink jet 3D printer, for example, the ink cartridge of the present invention is accommodated in the ink accommodating portion. The modeling of the three-dimensional model can be appropriately performed according to, for example, the type of the ink jet 3D printer. Specifically, the three-dimensional data is input to the ink jet 3D printer and the model is modeled with default settings. Can be implemented. The ink jet 3D printer may be, for example, an ink jet 3D printer capable of ejecting one type of ink or an ink jet 3D printer capable of ejecting two or more types of ink. As the cell ink, for example, a cell ink obtained by the cell ink production method of the present invention may be used instead of the cell ink of the present invention.
　また、前記立体造形工程において、前記インクジェット式３Ｄプリンターは、前記細胞インクに加え、他のインクを前記インク吐出口から吐出し、前記立体造形物を造形してもよい。前記他のインクは、例えば、前記細胞塊以外の成分を含むインクがあげられ、具体的には、ゲル等を含むインク、調味料等を含むインク、水溶性ポリマーを含むインク、前記刺激応答性分子を含むインク、前記細胞外マトリックスを含むインク等があげられる。 Further, in the three-dimensional modeling step, the ink jet 3D printer may model the three-dimensional model by discharging other ink from the ink discharge port in addition to the cell ink. Examples of the other ink include an ink containing a component other than the cell mass. Specifically, an ink containing a gel or the like, an ink containing a seasoning, an ink containing a water-soluble polymer, the stimulus responsiveness, and the like. Examples thereof include ink containing molecules and ink containing the extracellular matrix.
　本発明の立体造形物の製造方法において、例えば、複数の組織を含む複雑な臓器等を造形できることから、前記造形対象物が、複数の組織を含み、前記細胞インクが、各組織に対応する複数の細胞インクを含み、前記立体造形工程において、前記複数の細胞インクを用いて、前記複数の組織の情報を含む造形対象物の立体データに基づき、インクを吐出して立体造形を行う立体造形装置のインク吐出口から、前記細胞インクを吐出し、複数の組織を含む立体造形物を造形することが好ましい。 In the method for manufacturing a three-dimensional modeled object of the present invention, for example, a complex organ including a plurality of tissues can be modeled. Therefore, the modeling object includes a plurality of tissues, and the cell ink corresponds to each tissue. 3D modeling apparatus for performing 3D modeling by discharging ink based on 3D data of a modeling object including information on the plurality of tissues using the plurality of cell inks in the 3D modeling process Preferably, the cellular ink is discharged from the ink discharge port to form a three-dimensional structure including a plurality of tissues.
　具体的には、前記造形対象物が血管組織、脂肪組織、および筋肉組織を含む造形対象物の場合、例えば、以下のように実施できる。なお、これらは一例であり、本発明を制限するものではない。 Specifically, when the modeling object is a modeling object including a vascular tissue, a fat tissue, and a muscle tissue, for example, it can be performed as follows. Note that these are examples and do not limit the present invention.
　まず、各組織に含まれる細胞から構成される細胞塊を含む細胞インクを、前記本発明の細胞インクの製造方法により製造する。前記血管組織に対応する細胞インクは、例えば、血管内皮細胞等を含む。前記脂肪組織に対応する細胞インクは、例えば、脂肪細胞等を含む。前記筋肉組織に対応するインクは、例えば、筋細胞等を含む。各細胞インクにおいて、各細胞塊の大きさは、例えば、前記造形対象物における各組織の占める割合に応じて、設定する。そして、前記細胞インクを、前記インクジェット式３Ｄプリンターに収容されるインクカートリッジに収容する。 First, a cell ink containing a cell mass composed of cells contained in each tissue is produced by the cell ink production method of the present invention. The cell ink corresponding to the vascular tissue includes, for example, vascular endothelial cells. The cell ink corresponding to the fat tissue includes, for example, fat cells. The ink corresponding to the muscle tissue includes, for example, muscle cells. In each cell ink, the size of each cell mass is set according to, for example, the proportion of each tissue in the modeling object. And the said cell ink is accommodated in the ink cartridge accommodated in the said inkjet type 3D printer.
　つぎに、前記インクジェット式３Ｄプリンターのインク収容部に、各細胞インクを含む前記インクカートリッジを収容する。また、前記インクジェット式３Ｄプリンターで、前記血管組織、前記脂肪組織、および前記筋肉組織の情報を含む立体データを取得する。そして、前記各組織に対応する細胞インクを、前記各組織に対応する位置に吐出するよう設定する。なお、前記造形対象物が空隙を有する場合、例えば、前記空隙に対し、前記各細胞インクを吐出しないように、前記インクジェット式３Ｄプリンターを設定してもよいし、前記ゲル等を含むその他のインクを吐出するように、前記インクジェット式３Ｄプリンターを設定してもよい。前記空隙は、例えば、血管組織内の空間等があげられる。 Next, the ink cartridge containing each cell ink is accommodated in the ink accommodating portion of the ink jet 3D printer. Further, the ink jet 3D printer acquires three-dimensional data including information on the vascular tissue, the fat tissue, and the muscle tissue. And it sets so that the cell ink corresponding to each said tissue may be discharged to the position corresponding to each said tissue. In addition, when the said modeling target object has a space | gap, you may set the said inkjet type 3D printer so that each said cell ink may not be discharged with respect to the said space | gap, for example, and other ink containing the said gel etc. The ink jet 3D printer may be set so as to eject the ink. Examples of the void include a space in vascular tissue.
　そして、前記立体データに基づき、前記インクジェット式３Ｄプリンターが、前記各組織に対応する位置に前記各細胞インクを吐出することにより、前記各組織を含む前記立体造形物を造形する。なお、前記血管組織に対応する細胞インクとして、血管内皮細胞等を含む細胞インクを吐出する場合を例にあげたが、例えば、前記血管組織に対応する細胞インクに代えて、前記血管組織に分化する前駆細胞を含む細胞インク、前記刺激応答性分子を含むインク、前記細胞外マトリックスを含むインク等を用いて、前記血管組織を造形してもよい。 Then, based on the three-dimensional data, the inkjet 3D printer ejects each cell ink to a position corresponding to each tissue, thereby modeling the three-dimensional structure including each tissue. In addition, although the case where the cell ink containing a vascular endothelial cell etc. is ejected as an example of the cell ink corresponding to the vascular tissue has been described as an example, for example, instead of the cell ink corresponding to the vascular tissue, differentiation into the vascular tissue The vascular tissue may be shaped using a cell ink containing progenitor cells, an ink containing the stimulus-responsive molecule, an ink containing the extracellular matrix, or the like.
　このようにして、前記立体造形物を造形することができる。 In this way, the three-dimensional model can be modeled.
＜立体造形物＞
　本発明の立体造形物は、前述のように、前記本発明の立体造形物の製造方法により得られることを特徴とする。本発明の立体造形物は、前記本発明の立体造形物の製造方法により得られたことが特徴であり、その他の構成および条件は、特に制限されない。本発明の立体造形物は、例えば、前記本発明の細胞インク等の説明を援用できる。 <3D objects>
As described above, the three-dimensional object of the present invention is obtained by the method for manufacturing a three-dimensional object of the present invention. The three-dimensional modeled object of the present invention is obtained by the method for manufacturing a three-dimensional modeled object of the present invention, and other configurations and conditions are not particularly limited. For example, the description of the cell ink of the present invention can be used for the three-dimensional modeled object of the present invention.
　以上、実施形態を参照して本発明を説明したが、本発明は、上記実施形態に限定されるものではない。本発明の構成や詳細には、本発明のスコープ内で当業者が理解しうる様々な変更をできる。 As mentioned above, although this invention was demonstrated with reference to embodiment, this invention is not limited to the said embodiment. Various changes that can be understood by those skilled in the art can be made to the configuration and details of the present invention within the scope of the present invention.
　この出願は、２０１５年８月２４日に出願された日本出願特願２０１５－１６５２９５を基礎とする優先権を主張し、その開示のすべてをここに取り込む。 This application claims priority based on Japanese Patent Application No. 2015-165295 filed on August 24, 2015, the entire disclosure of which is incorporated herein.
　本発明によれば、立体造形物の造形が可能な細胞インクを製造可能である。このため、本発明は、例えば、医療分野、食品分野等において、極めて有用である。 According to the present invention, it is possible to produce a cell ink capable of modeling a three-dimensional model. For this reason, this invention is very useful in the medical field, the foodstuff field, etc., for example.
１　　　　　　　　　容器
２　　　　　　　　　底部
３　　　　　　　　　開口部
４　　　　　　　　　側壁
５　　　　　　　　　縁
６　　　　　　　　　保持部
１０　　　　　　　　製造容器
１１　　　　　　　　回収容器
２０　　　　　　　　製造キット DESCRIPTION OF SYMBOLS 1 Container 2 Bottom part 3 Opening part 4 Side wall 5 Edge 6 Holding part 10 Manufacturing container 11 Recovery container 20 Manufacturing kit

Claims (34)

  1. 細胞塊を含むことを特徴とする、細胞インク。 A cell ink comprising a cell mass.
  2. 前記細胞塊が、均一な細胞塊である、請求項１記載の細胞インク。 The cell ink according to claim 1, wherein the cell mass is a uniform cell mass.
  3. 前記細胞インクに含まれる細胞塊において、９０％以上の細胞塊が、所定径に対して±３０％の範囲の径を有する、請求項１または２記載の細胞インク。 3. The cell ink according to claim 1, wherein in the cell mass contained in the cell ink, 90% or more of the cell masses have a diameter in a range of ± 30% with respect to a predetermined diameter.
  4. 前記細胞塊の径が、インクを吐出して立体造形を行う立体造形装置のインク吐出口の径の大きさ以下である、請求項１から３のいずれか一項に記載の細胞インク。 The cell ink according to any one of claims 1 to 3, wherein a diameter of the cell mass is equal to or less than a diameter of an ink discharge port of a three-dimensional modeling apparatus that performs three-dimensional modeling by discharging ink.
  5. 前記細胞塊の径が、インクを吐出して立体造形を行う立体造形装置のインク吐出口の径に対して１／４～１倍の範囲の長さである、請求項１から４のいずれか一項に記載の細胞インク。 The diameter of the cell mass is a length in a range of 1/4 to 1 times the diameter of an ink discharge port of a three-dimensional modeling apparatus that performs three-dimensional modeling by discharging ink. The cell ink according to one item.
  6. 前記細胞塊を構成する細胞の細胞表面におけるインテグリンおよびカドヘリンの少なくとも一方の密度が損なわれていない、請求項１から５のいずれか一項に記載の細胞インク。 The cell ink according to any one of claims 1 to 5, wherein the density of at least one of integrin and cadherin on the cell surface of the cells constituting the cell mass is not impaired.
  7. 有底筒状の容器を含み、
    前記容器の底部は、１以上の開口部を含み、
    前記開口部は、細胞塊を培養するための培養室であり、
    前記底部の外面側における前記開口部の径の大きさが、インクを吐出して立体造形を行う立体造形装置のインク吐出口の径の大きさ以下であることを特徴とする、細胞インクの製造容器。     Including a bottomed cylindrical container,
    The bottom of the container includes one or more openings;
    The opening is a culture chamber for culturing a cell mass,
    Manufacturing of cellular ink, wherein the size of the diameter of the opening on the outer surface side of the bottom is equal to or less than the size of the diameter of an ink ejection port of a three-dimensional modeling apparatus that performs three-dimensional modeling by discharging ink container.
  8. 前記外面側における前記開口部の径が、５０～１０００μｍである、請求項７記載の細胞インクの製造容器。 The cell ink production container according to claim 7, wherein the diameter of the opening on the outer surface side is 50 to 1000 袖 m.
  9. 前記開口部が、前記底部の内面側から外面側に向かって、テーパー状である、請求項７または８記載の細胞インクの製造容器。 The cell ink production container according to claim 7 or 8, wherein the opening is tapered from the inner surface side to the outer surface side of the bottom portion.
  10. 前記開口部の径が、前記底部の内面側から外面側に向かって、減少する、請求項７から９のいずれか一項に記載の細胞インクの製造容器。 The cell ink production container according to any one of claims 7 to 9, wherein a diameter of the opening portion decreases from an inner surface side to an outer surface side of the bottom portion.
  11. 前記開口部の高さが、所望の径を有する細胞塊の半径Ｒに対して１～１０倍の高さである、請求項７から１０のいずれか一項に記載の細胞インクの製造容器。 The cell ink production container according to any one of claims 7 to 10, wherein the height of the opening is 1 to 10 times the radius R of a cell mass having a desired diameter.
  12. 請求項７から１１のいずれか一項に記載の細胞インクの製造容器と、細胞塊を回収するための回収容器とを含み、
    前記回収容器は、有底筒状であり、内部に前記細胞インクの製造容器を収容可能であることを特徴とする、細胞インクの製造キット。     A cell ink production container according to any one of claims 7 to 11, and a collection container for collecting cell masses,
    The cell ink production kit, wherein the collection container has a bottomed cylindrical shape and can accommodate the cell ink production container.
  13. 前記回収容器は、前記細胞インクの製造容器を内部に保持する保持部を含み、
    前記保持部は、前記回収容器の底部の内面と、前記細胞インクの製造容器の底部の外面とを、一定距離を保つように前記回収容器に配置される、請求項１２記載の細胞インクの製造キット。     The collection container includes a holding unit that holds the cell ink production container inside,
    The said holding | maintenance part is manufactured in the said collection | recovery container so that the inner surface of the bottom part of the said collection | recovery container and the outer surface of the bottom part of the said production | generation container of a cell ink may maintain a fixed distance. kit.
  14. 前記回収容器が、インクを吐出して立体造形を行う立体造形装置用のインクタンクであり、前記回収容器が、インク吐出部を含む、請求項１２または１３記載の細胞インクの製造キット。 The cellular ink manufacturing kit according to claim 12 or 13, wherein the collection container is an ink tank for a three-dimensional modeling apparatus that performs three-dimensional modeling by discharging ink, and the collection container includes an ink discharge unit.
  15. さらに、前記細胞インクの製造容器または前記回収容器の開口を塞ぐ蓋を含む、請求項１２から１４のいずれか一項に記載の細胞インクの製造キット。 The cell ink production kit according to claim 12, further comprising a lid that closes an opening of the cell ink production container or the collection container.
  16. 請求項７から１１のいずれか一項に記載の細胞インクの製造容器を用いて細胞を培養する培養工程、および
    培養された細胞塊を回収する回収工程を含み、
    前記培養工程において、培養液の存在下、前記細胞インクの製造容器の前記開口部において、細胞を培養し、
    前記回収工程において、前記開口部から、前記細胞インクの製造容器の外部に導出された細胞塊を回収することを特徴とする、細胞インクの製造方法。     A culture step of culturing cells using the cell ink production container according to any one of claims 7 to 11, and a recovery step of collecting the cultured cell mass,
    In the culturing step, cells are cultured in the opening of the cell ink production container in the presence of a culture solution,
    In the collection step, a cell mass derived from the opening to the outside of the cell ink production container is collected.
  17. 前記培養工程において、
    前記細胞インクの製造容器を含む請求項１２から１５のいずれか一項に記載の細胞インクの製造キットを使用し、
    前記細胞インクの製造容器を前記回収容器の内部に収容し、前記培養液の存在下、細胞を培養し、
    前記回収工程において、前記開口部から、前記細胞インクの製造容器の外部の前記回収容器に導出された細胞塊を回収する、請求項１６記載の細胞インクの製造方法。     In the culturing step,
    A cell ink production kit according to any one of claims 12 to 15, comprising the cell ink production container,
    Storing the cell ink production container in the collection container, culturing the cells in the presence of the culture solution,
    The method for producing cell ink according to claim 16, wherein in the collecting step, the cell mass led to the collecting container outside the cell ink producing container is collected from the opening.
  18. 前記回収工程において、前記回収容器に回収溶液を導入し、前記開口部と前記回収溶液とを接触させることにより、前記開口部から、前記細胞インクの製造容器の外部の前記回収容器に導出された細胞塊を回収する、請求項１７記載の細胞インクの製造方法。 In the recovery step, the recovery solution is introduced into the recovery container, and the opening and the recovery solution are brought into contact with each other to be led out from the opening to the recovery container outside the cell ink manufacturing container. The method for producing a cell ink according to claim 17, wherein the cell mass is collected.
  19. 前記回収溶液が、細胞インク用溶液である、請求項１８記載の細胞インクの製造方法。 19. The method for producing cell ink according to claim 18, wherein the recovered solution is a cell ink solution.
  20. 前記回収工程において、前記回収容器から前記細胞インクの製造容器を取り外し、前記回収容器の内部の前記細胞塊を回収する、請求項１７から１９のいずれか一項に記載の細胞インクの製造方法。 The method for producing cell ink according to any one of claims 17 to 19, wherein, in the collecting step, the cell ink producing container is removed from the collecting container, and the cell mass inside the collecting container is collected.
  21. 前記回収工程に先立ち、前記培養液を細胞インク用溶液と交換する溶液交換工程を含む、請求項１６から２０のいずれか一項に記載の細胞インクの製造方法。 The method for producing cell ink according to any one of claims 16 to 20, further comprising a solution exchanging step of exchanging the culture solution with a cell ink solution prior to the collecting step.
  22. 前記細胞インク用溶液が、脂質、細胞外マトリックス、生体材料、および刺激応答性分子からなる群から選択された少なくとも１つを含む、請求項１９または２１記載の細胞インクの製造方法。 The method for producing cell ink according to claim 19 or 21, wherein the solution for cell ink comprises at least one selected from the group consisting of lipid, extracellular matrix, biomaterial, and stimulus-responsive molecule.
  23. 前記培養開始時において、前記細胞が分散状態である、請求項１６から２２のいずれか一項に記載の細胞インクの製造方法。 The method for producing cell ink according to any one of claims 16 to 22, wherein the cells are in a dispersed state at the start of the culture.
  24. 請求項１６から２３のいずれか一項に記載の細胞インクの製造方法により得られることを特徴とする、細胞インク。 24. A cell ink obtained by the method for producing a cell ink according to any one of claims 16 to 23.
  25. 請求項１から６および２４のいずれか一項に記載の細胞インクを含むことを特徴とする、インクを吐出して立体造形を行う立体造形装置用のインクカートリッジ。 An ink cartridge for a three-dimensional modeling apparatus for performing three-dimensional modeling by discharging ink, comprising the cell ink according to any one of claims 1 to 6 and 24.
  26. 細胞インクを用いる立体造形工程を含み、
    前記立体造形工程が、造形対象物の立体データに基づき、インクを吐出して立体造形を行う立体造形装置のインク吐出口から、前記細胞インクを吐出し、立体造形物を造形し、
    前記細胞インクが、請求項１から６のいずれか一項に記載の細胞インク、および請求項１６から２３のいずれか一項に記載の細胞インクの製造方法により得られた細胞インクの少なくとも一方であることを特徴とする、立体造形物の製造方法。     Including a three-dimensional modeling process using cell ink,
    The three-dimensional modeling step is based on the three-dimensional data of the modeling object, and discharges the cell ink from the ink discharge port of the three-dimensional modeling apparatus that performs ink three-dimensional modeling by discharging ink, and models the three-dimensional modeling object,
    The cell ink according to any one of claims 1 to 6 and the cell ink obtained by the method for producing cell ink according to any one of claims 16 to 23 are used as the cell ink. A method for producing a three-dimensional structure, characterized by being.
  27. さらに、前記細胞インクを製造する細胞インクの製造工程を含み、
    前記細胞インクの製造工程が、請求項１６から２３のいずれか一項に記載の細胞インクの製造方法により実施される、請求項２６記載の立体造形物の製造方法。     And further comprising a cell ink production process for producing the cell ink,
    The manufacturing method of the three-dimensional molded item according to claim 26, wherein the manufacturing process of the cell ink is performed by the manufacturing method of the cell ink according to any one of claims 16 to 23.
  28. さらに、前記造形対象物の立体データを取得する立体データ取得工程を含む、請求項２６または２７記載の立体造形物の製造方法。 Furthermore, the manufacturing method of the three-dimensional molded item of Claim 26 or 27 including the three-dimensional data acquisition process of acquiring the three-dimensional data of the said modeling target object.
  29. 前記立体データが、前記造形対象物を構成する組織の情報を含む、請求項２６から２８のいずれか一項に記載の立体造形物の製造方法。 The manufacturing method of the three-dimensional molded item according to any one of claims 26 to 28, wherein the three-dimensional data includes information on a structure constituting the modeling target object.
  30. 前記組織情報が、上皮組織、結合組織、神経組織、血管組織、脂肪組織、および筋肉組織からなる群から選択された少なくとも１つの組織情報である、請求項２９記載の立体造形物の製造方法。 30. The method for producing a three-dimensional structure according to claim 29, wherein the tissue information is at least one tissue information selected from the group consisting of epithelial tissue, connective tissue, nerve tissue, vascular tissue, adipose tissue, and muscle tissue.
  31. 前記造形対象物が、血管を含む造形対象物であり、
    前記立体データ取得工程において、前記造形対象物の血管組織の情報を含む立体データを取得する、請求項２８または２９記載の立体造形物の製造方法。     The modeling object is a modeling object including a blood vessel,
    30. The method of manufacturing a three-dimensional structure according to claim 28 or 29, wherein in the three-dimensional data acquisition step, three-dimensional data including information on a vascular tissue of the modeling object is acquired.
  32. 前記立体データが、三次元ＣＴ画像、三次元ＭＲＩ画像、三次元ＰＥＴ画像、または三次元ＰＥＴ－ＣＴ画像である、請求項２６から３１のいずれか一項に記載の立体造形物の製造方法。 32. The method of manufacturing a three-dimensional structure according to claim 26, wherein the three-dimensional data is a three-dimensional CT image, a three-dimensional MRI image, a three-dimensional PET image, or a three-dimensional PET-CT image.
  33. 前記造形対象物が、複数の組織を含み、
    前記細胞インクが、各組織に対応する複数の細胞インクを含み、
    前記立体造形工程において、前記複数の細胞インクを用いて、前記複数の組織の情報を含む造形対象物の立体データに基づき、インクを吐出して立体造形を行う立体造形装置のインク吐出口から、前記細胞インクを吐出し、複数の組織を含む立体造形物を造形する、請求項２６から３２のいずれか一項に記載の立体造形物の製造方法。     The modeling object includes a plurality of tissues,
    The cellular ink includes a plurality of cellular inks corresponding to each tissue,
    In the three-dimensional modeling step, using the plurality of cell inks, based on the three-dimensional data of the modeling object including information on the plurality of tissues, from the ink ejection port of the three-dimensional modeling apparatus that performs three-dimensional modeling by discharging ink, The method for producing a three-dimensional structure according to any one of claims 26 to 32, wherein the three-dimensional structure including a plurality of tissues is formed by discharging the cell ink.
  34. 請求項２６から３３のいずれか一項に記載の立体造形物の製造方法により得られることを特徴とする、立体造形物。 It is obtained by the manufacturing method of the three-dimensional molded item as described in any one of Claim 26 to 33, The three-dimensional molded item characterized by the above-mentioned.
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