# US 12,215,357 B2 · Apparatus and method for establishment of induced pluripotent stem cells > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 12,215,357 B2 - Document ID: US12215357B2 - Status: Granted - Kind: B2 - Application number: 17/503,634 - Filed: 2021-10-18 - Published: 2022-04-21 - Granted: 2025-02-04 - Earliest claimed: 2020-10-19 - Assignees: Canon Medical Systems Corporation; Canon Kabushiki Kaisha - Applicants: Canon Medical Systems Corporation; Canon Kabushiki Kaisha - Inventors: Katsuaki Deguchi - Web page: https://patentgrep.com/patent/US12215357B2 ## CPC - C12N 5/0696: {Artificially induced pluripotent stem cells, e.g. iPS} - C12M 33/14: {with filters, sieves or membranes} - C12M 35/08: {Chemical, biochemical or biological means, e.g. plasma jet, co-culture} - C12M 45/06: {by chemical means or hydrolysis} - C12M 47/04: {Cell isolation or sorting (purging biological preparations of unwanted cells C12N5/0081, determining the presence or kind of microorganism C12Q1/04)} - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) - 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} ## Abstract According to one embodiment, a method for the establishment of an induced pluripotent stem (iPS) cell includes a suspension liquid feeding step, an inducing factor feeding step, and an establishing step. The suspension liquid feeding step feeds a suspension liquid containing a target cell. The inducing factor feeding step feeds an inducing factor to a trap. The trap traps the target cell contained in the suspension liquid fed in the suspension liquid feeding step. The establishing step establishes an iPS cell by introducing the inducing factor, which has been fed to the trap, into the target cell trapped by the trap. ## Claims ### Claim 1 A method for supplying a mixed liquid containing a mononuclear cell and an inducing factor for inducing reprogramming of the mononuclear cell to a container, the method comprising: a first step of sending a suspension liquid containing the mononuclear cell to a trap through a first flow channel in a first direction, the trap configured to trap the mononuclear cell contained in the suspension liquid sent in the first step, the trap being arranged in the first flow channel; and a second step of sending the inducing factor to the trap in a second direction opposite the first direction through the first flow channel, and sending the mononuclear cell trapped by the trap and the inducing factor to the container through a second flow channel branching off from the first flow channel at a branch position upstream of the trap in the first direction. ### Claim 2 (depends on 1) The method according to claim 1, wherein the trap comprises a mesh to catch the mononuclear cell, the mesh being configured to catch a substance having a particle size larger than an opening of the mesh and permit a substance having a particle size smaller than the opening to pass through. ### Claim 3 (depends on 1) The method according to claim 1, wherein the trap comprises a first mesh and a second mesh, the first mesh being configured to catch a substance having a first particle size larger than the mononuclear cell and permit a substance having a particle size smaller than the first particle size to pass through, the second mesh being configured to catch a substance that has passed through the first mesh and has a second particle size equivalent to the mononuclear cell, and permit a substance having a particle size smaller than the second particle size to pass through. ### Claim 4 (depends on 3) The method according to claim 3, wherein the second step sends the inducing factor to a space between the first mesh and the second mesh. ### Claim 5 (depends on 1) The method according to claim 1, further comprising a wash liquid feeding step of sending a wash liquid to the trap, by which a substance other than the mononuclear cell remaining in the trap is washed away. ### Claim 6 (depends on 5) The method according to claim 5, wherein the wash liquid feeding step comprises sending the wash liquid to the trap after the first step sends the suspension liquid and before the second step sends the inducing factor. ### Claim 7 (depends on 5) The method according to claim 5, wherein the wash liquid feeding step comprises repeatedly suctioning and discharging the wash liquid sent to the trap so that the substance other than the mononuclear cell remaining in the trap is washed away. ### Claim 8 (depends on 1) The method according to claim 1, wherein the second step sends, together with the inducing factor, a culture medium. ### Claim 9 (depends on 1) The method according to claim 1, wherein the first step sends blood or a preservative solution as the suspension liquid. ### Claim 10 (depends on 1) The method according to claim 1, further comprising: sending a preservative solution to the trap, and recovering the preservative solution containing the mononuclear cell trapped by the trap. ### Claim 11 A liquid supplying apparatus, comprising: a suspension liquid feeder configured to send a suspension liquid containing a mononuclear cell through a first flow channel in a first direction: a trap connected to the suspension liquid feeder via the first flow channel and configured to trap the mononuclear cell contained in the suspension liquid sent through the first flow channel from the suspension liquid feeder, the trap being arranged in the first flow channel; and an inducing factor feeder, the inducing factor feeder being configured to send an inducing factor for inducing reprogramming of the mononuclear cell to the trap in a second direction opposite the first direction through the first flow channel, and to send a liquid containing the mononuclear cell and the inducing factor to a container, the container being connected to the trap via a second flow channel branching off from the first flow channel at a branch position upstream of the trap in the first direction. ### Claim 12 (depends on 11) The apparatus according to claim 11, wherein the trap comprises a mesh to catch the mononuclear cell, the mesh being configured to catch a substance having a particle size larger than an opening of the mesh and permit a substance having a particle size smaller than the opening to pass through. ### Claim 13 (depends on 11) The apparatus according to claim 11, wherein the trap comprises a first mesh and a second mesh, the first mesh being configured to catch a substance having a first particle size larger than the mononuclear cell and permit a substance having a particle size smaller than the first particle size to pass through, the second mesh being configured to catch a substance that has passed through the first mesh and has a second particle size equivalent to the mononuclear cell, and permit a substance having a particle size smaller than the second particle size to pass through. ### Claim 14 (depends on 11) The apparatus according to claim 11, further comprising the first flow channel, into which the suspension liquid is introduced from the suspension liquid feeder and into which the inducing factor is introduced from the inducing factor feeder, wherein in the first flow channel, the suspension liquid flows in the first direction, and the inducing factor flows in the second direction opposite the first direction, and the trap is located in the first flow channel. ### Claim 15 (depends on 14) The apparatus according to claim 14, wherein an establisher is connected to the second flow channel so that the mononuclear cell and the inducing factor are sent to the establisher via the second flow channel. ### Claim 16 (depends on 11) The apparatus according to claim 11, further comprising a controller configured to control the trap, wherein, at a time of feeding the suspension liquid from the suspension liquid feeder, the controller is configured to control the trap to catch the mononuclear cell contained in the suspension liquid, and at a time of feeding the inducing factor from the inducing factor feeder, the controller is configured to control the trap to release the mononuclear cell.