# US 2021/0277345 A1 · Cell-containing container, method for evaluating test substance, and method for manufacturing cell-containing container > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 2021/0277345 A1 - Document ID: US20210277345A1 - Status: Published application - Kind: A1 - Application number: 17/190,487 - Filed: 2021-03-03 - Published: 2021-09-09 - Earliest claimed: 2020-03-06 - Inventors: Tomoyuki ARATANI; Daisuke TAKAGI; Hidekazu YAGINUMA; Waka LIN; Shinnosuke KOSHIZUKA; Ryuya MASHIKO; Tomoaki NAKAYAMA; Atsushi MIYAOKA; Takeru SUZUKI; Minoru KO - Web page: https://patentgrep.com/patent/US20210277345A1 ## CPC - C12M 23/04: {Flat or tray type, drawers (C12M23/10, C12M23/12, C12M23/16 take precedence)} - C12M 23/12: {Well or multiwell plates (C12M25/04 takes precedence)} - C12M 25/02: {Membranes; Filters (filters or filtration in general B01D24/00-B01D41/00)} - C12M 33/00: {Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus (chemical or physical laboratory apparatus in general B01L, devices for taking cell samples A61B10/0045, withdrawing or distributing predetermined quantities of fluid B01L99/00)} - C12M 35/08: {Chemical, biochemical or biological means, e.g. plasma jet, co-culture} - C12M 41/46: {of cellular or enzymatic activity or functionality, e.g. cell viability} - C12N 5/0607: {Non-embryonic pluripotent stem cells, e.g. MASC (induced pluripotent stem cells C12N5/0696)} - C12N 5/0696: {Artificially induced pluripotent stem cells, e.g. iPS} - C12N 2535/00: Supports or coatings for cell culture characterised by topography - C12M: APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; {APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS} - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) ## Abstract A cell-containing container includes at least one recessed part accommodating cells, in which the cells adhere to a bottom surface of the recessed part, and a cell density in a first region of the bottom surface is 250 to 20,000 cells/mm2, and a cell density in a second region surrounding a periphery of the first region is less than 250 cells/mm2. ## Claims ### Claim 1 A cell-containing container, comprising: at least one recessed part accommodating cells, wherein the cells adhere to a bottom surface of the recessed part, and a cell density in a first region of the bottom surface is 250 to 20,000 cells/mm2, and a cell density in a second region surrounding a periphery of the first region is less than 250 cells/mm2. ### Claim 2 (depends on 1) The cell-containing container according to claim 1, wherein a cell density in the first region is 500 to 10,000 cells/mm2. ### Claim 3 (depends on 1) The cell-containing container according to claim 1, wherein the cells are primary cells, stem cells or induced pluripotent stem (iPS) cells, wherein the cells are derived from a patient or a healthy subject, or cells differentiated from the iPS cells. ### Claim 4 (depends on 1) The cell-containing container according to claim 1, comprising: a plurality of recessed parts, wherein each of the recessed parts accommodates cells derived from the same patient or the same healthy subject, and the plurality of recessed parts include recessed parts each accommodating cells derived from different patients or different healthy subjects. ### Claim 5 (depends on 1) The cell-containing container according to claim 1, wherein a degree of aggregation of cells in the first region is 100% or less, which is calculated by Formula (1): degree of aggregation (%)=A/B×100 (1) [in Formula (1), A indicates a standard deviation of area of Voronoi regions of each cells, and B indicates an average value of areas of Voronoi regions of each cells, where a Voronoi region of a cell is a region surrounded by perpendicular bisectors in a case where a perpendicular bisector is drawn between a nucleus of a cell and a nucleus of a cell adjacent to the cell]. ### Claim 6 (depends on 1) The cell-containing container according to claim 1, wherein the recessed part accommodates (i) nerve cells, and (ii) at least one cells selected from the group consisting of glial cells, vascular endothelial cells, and smooth muscle cells. ### Claim 7 (depends on 1) The cell-containing container according to claim 1, wherein the recessed part accommodates nerve cells, and an electrode array is provided on the bottom surface of the recessed part. ### Claim 8 A method for evaluating a test substance, the method comprising: adding a test substance into a recessed part of a cell-containing container which has at least one recessed part containing cells and in which a cell density in a center of a bottom surface of the recessed part is higher than that in an edge portion of the bottom surface of the recessed part; and detecting spontaneous firing of nerve cells through an electrode array provided on the bottom surface of the recessed part. ### Claim 9 A method for manufacturing a cell-containing container, the method comprising controlling the number and positions of cells and jetting the cells by an inkjet technique, where the controlling is performed such that, in a recessed part of a container having at least one recessed part, a cell density in a center of a bottom surface of the recessed part is 250 to 20,000 cells/mm2, and a cell density in an edge portion of the bottom surface of the recessed part is less than 250 cells/mm2. ### Claim 10 (depends on 9) The method for manufacturing a cell-containing container according to claim 9, wherein jetting the cells is performed by an inkjet head including at least a liquid-holding part which holds a cell suspension containing cells, a film-like member on which a nozzle is formed and which jets, from the nozzle, the cell suspension held in the liquid-holding part as liquid droplets by vibration, and an atmospheric opening part which opens an inside of the liquid-holding part to the atmosphere. ### Claim 11 (depends on 10) The method for manufacturing a cell-containing container according to claim 10, wherein jetting the cells is performed by operating a plurality of inkjet heads simultaneously or alternatively.