Search

CN-121986155-A - Cell separation device and cell separation method

CN121986155ACN 121986155 ACN121986155 ACN 121986155ACN-121986155-A

Abstract

The cell separation device of the embodiment is provided with a culture container and a discharge unit. The culture vessel cultures the cells. The ejection unit is provided at a position not immersed in the liquid in the culture container, and ejects a plurality of droplets onto a culture surface of the culture container to peel the cells from the culture surface.

Inventors

  • IKEDA SHUHEI
  • EBISAWA HISAFUMI
  • UCHIDA KOICHI

Assignees

  • 佳能株式会社
  • 佳能医疗系统株式会社

Dates

Publication Date
20260505
Application Date
20240920
Priority Date
20230922

Claims (20)

  1. 1. A cell separation device is provided with: Culture vessel for culturing cells, and And a discharge unit which is provided in the culture container at a position not immersed in the liquid and discharges a plurality of droplets toward the culture surface of the culture container, thereby peeling the cells from the culture surface.
  2. 2. The cell separation device according to claim 1, wherein, The ejection section ejects the plurality of droplets to a portion of the surface of the cell in contact with air.
  3. 3. The cell separation device according to claim 1 or claim 2, wherein, The plurality of droplets have an average particle diameter of 100 μm or more and 500 μm or less.
  4. 4. The cell separation device according to any one of claim 1 to claim 3, wherein, The average speed of the plurality of droplets is 5km/h or more and 50km/h or less.
  5. 5. The cell separation device according to any one of claim 1 to claim 4, wherein, The average kinetic energy of the plurality of droplets is 5.0X10 -13 J or more and 6.3X10 -9 J or less.
  6. 6. The cell separation device according to any one of claim 1 to claim 5, wherein, The total discharge amount of the plurality of droplets discharged at one time is 3.0mL or less.
  7. 7. The cell separation device according to any one of claim 1 to claim 6, wherein, The discharge time of the plurality of droplets discharged at one time is 0.2 seconds or less.
  8. 8. The cell separation device according to any one of claim 1 to claim 7, wherein, The distance between the culture surface and the ejection part is 10mm to 50 mm.
  9. 9. The cell separation device according to any one of claims 1 to 8, further comprising: A storage part connected with the ejection part for storing the liquid, and A transfer unit for transferring kinetic energy to the liquid stored in the storage unit, The liquid stored in the storage portion is ejected as the plurality of droplets from the ejection portion in a state where the kinetic energy is transmitted by the transmission portion.
  10. 10. The cell separation device according to claim 9, wherein, The storage portion is a hollow cylindrical body having one end connected to the discharge portion, and the transmission portion is a plunger that is attached to the inside of the cylindrical body and moves the liquid toward the one end.
  11. 11. The cell separation device according to any one of claim 1 to claim 10, wherein, Further comprising a discharge unit for discharging the liquid from the culture vessel, The discharge unit discharges liquid from the culture container to reduce the liquid amount in the culture container, and the discharge unit discharges the plurality of droplets to the expanded contact portion of the surface of the cells, in addition to expanding the contact portion with air.
  12. 12. A cell peeling method comprising the steps of: The cells cultured in the culture vessel are peeled off from the culture surface by ejecting a plurality of droplets toward the culture surface of the culture vessel from a position not immersed in the liquid in the culture vessel.
  13. 13. The method for cell separation according to claim 12, wherein, The plurality of droplets are ejected to a contact portion with air in the surface of the cell.
  14. 14. The cell separation method according to claim 12 or claim 13, wherein, The plurality of droplets have an average particle diameter of 100 μm or more and 500 μm or less.
  15. 15. The cell separation method according to any one of claim 12 to claim 14, wherein, The average speed of the plurality of droplets is 5km/h or more and 50km/h or less.
  16. 16. The cell separation method according to any one of claim 12 to claim 15, wherein, The average kinetic energy of the plurality of droplets is 5.0X10 -13 J or more and 6.3X10 -9 J or less.
  17. 17. The cell separation method according to any one of claim 12 to claim 16, wherein, The total discharge amount of the plurality of droplets discharged at one time is 3.0mL or less.
  18. 18. The cell separation method according to any one of claim 12 to claim 17, wherein, The discharge time of the plurality of droplets discharged at one time is 0.2 seconds or less.
  19. 19. The cell separation method according to any one of claim 12 to claim 18, wherein, The distance between the culture surface and the position is 10mm or more and 50mm or less.
  20. 20. The cell separation method according to any one of claim 12 to claim 19, wherein, The liquid amount in the culture vessel is reduced by discharging the liquid from the culture vessel, and the plurality of droplets are ejected to the enlarged contact portion of the surface of the cells with air.

Description

Cell separation device and cell separation method Technical Field Embodiments disclosed in the present specification and drawings relate to a cell separation apparatus and a cell separation method. Background In the cell culture process, it is necessary to peel off cells adhered to the bottom surface of the culture vessel. For example, a technician manually and physically peels off cells via a cell scraper or pipette. However, in the manual work, since there is a risk of contamination (contamination) of cells by microorganisms, damage (damage) to cells by mechanical stimulation, and the like, it is necessary to perform the peeling work by a skilled technician with careful attention while ensuring and maintaining a sterile environment. Prior art literature Patent literature Patent document 1 Japanese patent application laid-open No. 2011-155869 Disclosure of Invention Technical problem to be solved by the invention One of the technical problems to be solved by the embodiments disclosed in the present specification and the drawings is to easily peel off cells. However, the technical problems to be solved by the embodiments disclosed in the present specification and the drawings are not limited to the above technical problems. The technical problems corresponding to the effects of the respective configurations described in the embodiments described below can be also referred to as other technical problems. Means for solving the technical problems The cell separation device of the embodiment is provided with a culture container and a discharge unit. The culture vessel cultures the cells. The ejection unit is provided at a position not immersed in the liquid in the culture container, and ejects a plurality of droplets onto a culture surface of the culture container to peel the cells from the culture surface. Effects of the invention According to the present invention, cells can be easily detached. Drawings FIG. 1 is a schematic view showing an example of the structure of a cell separation device according to the first embodiment. FIG. 2 is a flowchart showing an example of the operation of the cell separation device according to the first embodiment. FIG. 3 is a schematic diagram showing an example of the operation of the cell separation device according to the first embodiment. FIG. 4 is a graph showing the results of cell separation for each experimental condition of the first embodiment. FIG. 5 is a schematic view showing an example of the structure of a cell separation device according to the second embodiment. FIG. 6 is a flowchart showing an example of the operation of the cell separation device according to the second embodiment. FIG. 7 is a schematic diagram showing an example of the operation of the cell separation device according to the second embodiment. Detailed Description Hereinafter, embodiments will be described with reference to the drawings. In the following embodiments, the same reference numerals are given to the same operations, and overlapping descriptions are appropriately omitted. (First embodiment) Fig. 1 is a schematic diagram showing an example of the structure of a cell separation device 1 according to the first embodiment. The figure shows the cell separation device 1 when viewed from the horizontal direction. The cell separation device 1 is a device for separating cells. The cell separation device 1 includes a culture container 2, a nozzle 3, a syringe 4, a pressing mechanism 5, a stopper 6, a support frame 7, a mount 8, a connector 9, and a control device 10. The culture vessel 2 is a vessel for culturing cells C. The culture vessel 2 is formed of a substance having a property (impermeability) of not penetrating the liquid L adhering to the surface. Examples of such materials include polyethylene, polypropylene, polycarbonate, polystyrene, and glass. The culture vessel 2 has an arbitrary size and an arbitrary shape. For example, the culture vessel 2 is a circular vessel having a diameter of 35mm or more and 100mm or less. The culture vessel 2 is an example of a culture section. The cells C are cells (adhesive cells) adhered to the surface of the culture vessel 2 via an adhesive protein. Examples of cell C include iPS (induced Pluripotent Stem) cells, epithelial cells, endothelial cells, synovial cells, cardiomyocytes, myoblasts, fibroblasts and neuroblasts. The cell C may be one cell or a colony (cell mass) composed of a plurality of cells. The liquid L is any kind of liquid. Examples of liquid L include PBS (Phosphate Buffered Saline) and liquid media. The liquid L is preferably a liquid which does not chemically or physiologically damage the cells C. The nozzle 3 is a mechanical member that ejects the liquid L. The nozzle 3 is formed of a relatively high-rigidity metal or the like. The nozzle 3 is physically connected to the syringe 4 by a connector 9. The distance d1 between the tip of the nozzle 3 and the bottom surface of the culture vessel 2 is preferably 10mm to 50 mm. The