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BR-112022019023-B1 - DEVICE FOR PERFORMING A CELL-BASED ASSAY AND CELL SEEDING, KIT FOR PERFORMING A CELL-BASED ASSAY, METHOD FOR PERFORMING A CELL-BASED ASSAY AND SEEDING CELLS ON THE SURFACE OF THE PIN

BR112022019023B1BR 112022019023 B1BR112022019023 B1BR 112022019023B1BR-112022019023-B1

Abstract

PIN, PIN SUPPORT, DEVICE FOR PERFORMING A CELL-BASED ASSAY, CELL SEEDING DEVICE, KIT FOR PERFORMING A CELL-BASED ASSAY, METHODS FOR PERFORMING A CELL-BASED ASSAY AND SEEDING CELLS ON THE PIN SURFACE. The present invention provides cell assay systems and their use in cell-based assays.

Inventors

  • Guojun Chen
  • Stephen Fowler
  • NA HONG QIU

Assignees

  • NCL NEW CONCEPT LAB GMBH
  • F. HOFFMANN-LA ROCHE AG

Dates

Publication Date
20260310
Application Date
20210322
Priority Date
20200323

Claims (20)

  1. 1. DEVICE FOR PERFORMING A CELL-BASED ASSAY (1a) comprising: a pin holder (2, 1307) comprising a plurality of openings (7a, 1314) for receiving a pin (1), and a base (3, 1301) comprising a groove channel (7, 1302), wherein the pin (1) comprises a pin body (4) comprising a pin head (4a) and a pin tip (6), wherein the pin tip (6) comprises a surface (6a) for seeding cells, wherein the pin holder (2, 1307) and the base (3, 1301) have a combined shape and the openings (7a, 1314) for receiving the pin (1) are arranged so that they are aligned with the groove channel (7, 1302) of the base (3, 1301), wherein the pin holder (1307) further comprises an opening (1312a) for receiving a cell culture insert (1312) and the base (1301) further comprising a reservoir for receiving a liquid (1303), wherein the reservoir (1303) is disposed within the base (1301), so that it is aligned with the opening (1312a) of the pin holder (1307) for receiving a cell culture insert (1312), characterized in that the device (1a) further comprises a pump (1311) which is fluidically connected to the reservoir (1303), to the groove channel (1302) and to the cell culture insert (1312).
  2. 2. DEVICE FOR PERFORMING A CELL-BASED ASSAY (1a), according to claim 1, characterized in that the pin support (1307) comprises: a plurality of openings (1314) for receiving a pin (1), at least two openings (1312a) for receiving at least two cell culture inserts (1312), at least one opening (1309) for receiving the pump impeller rod (1313), and the base (3, 1301) comprises: at least two reservoirs (1303) for receiving a liquid, in particular a cell culture medium, at least one opening (1304) for receiving the pump (1311), in particular a pump impeller (1311), an inlet channel (1305), an outlet channel (1306), wherein the inlet channel (1305) and the outlet channel (1306) of the pump (1313) are connected. to the reservoir (1303) and to the slot channel (1302).
  3. 3. DEVICE FOR PERFORMING A CELL-BASED TEST (1a) comprising: a pin holder (2, 1307) comprising a plurality of openings (7a, 1314) for receiving a pin (1), wherein the pin (1) comprises a pin body (4) comprising a pin head (4a) and a pin tip (6), wherein the pin tip (6) comprises a surface (6a) for seeding cells, and characterized by comprising a base (3) comprising a groove channel (7) and a reservoir (8), wherein the groove channel (7) is dimensioned to receive the pin tips (6) of the pins (1) being fixed in the openings (7a) of the pin holder (2), wherein the groove channel (7) is connected to the reservoir (8).
  4. 4. DEVICE FOR PERFORMING THE CELL-BASED TEST (1a), according to claim 3, characterized in that the pin support (2) and the base (3) are separable parts of the device.
  5. 5. DEVICE FOR PERFORMING THE CELL-BASED TEST (1a), according to claim 3, characterized in that the pin support (2) and the base (3) are integrated as a single piece.
  6. 6. DEVICE FOR PERFORMING THE CELL-BASED TEST (1a), according to any one of claims 3 to 5, characterized in that the base (2) comprises more than one groove channel (7).
  7. 7. DEVICE FOR PERFORMING THE CELL-BASED TEST (1a), according to any one of claims 3 to 6, characterized in that the groove channel (7) has a stepped shape comprising an upper part (11), a step (10) and a lower part (9), wherein the upper part (11) has a larger dimension than the lower part (9).
  8. 8. DEVICE FOR PERFORMING THE CELL-BASED TEST (1a), according to any one of claims 3 to 7, characterized in that the groove channel (7) is in a linear or circular arrangement in the base (3).
  9. 9. DEVICE FOR PERFORMING THE CELL-BASED TEST (1a), according to any one of claims 3 to 8, characterized in that the reservoir (8) contains the liquid.
  10. 10. DEVICE FOR PERFORMING THE CELL-BASED TEST (1a), according to any one of claims 3 to 9, characterized in that the pin support (2), the pins (1) and the base (3) are made of plastic, preferably polystyrene, polyester and polycarbonate.
  11. 11. CELL SEEDING DEVICE (40) for seeding cells on a pin surface (6a) of a pin (1), wherein the pin (1) comprises a pin body (4) comprising a pin head (4a) and a pin tip (6), wherein the pin tip (6) comprises a surface (6a) for seeding cells, characterized by comprising: a first part (41) and a second part (43) forming a plurality of funnel-shaped cavities (44) when assembled, wherein the funnel-shaped cavity (44) comprises an upper cavity part (44a) with an opening and a lower cavity part (44b) with an opening, wherein the lower cavity part (44b) of the funnel-shaped cavity (44) is dimensioned to receive the tip (6) of the pin (1).
  12. 12. CELL SEEDING DEVICE (40), according to claim 11, characterized in that the funnel-shaped cavities (44) are arranged in a die.
  13. 13. CELL SEEDING DEVICE (40), according to any one of claims 11 to 12, characterized in that the matrix is in a format of 8, 12, 16, 24 to 96 funnels.
  14. 14. CELL SEEDING DEVICE (40), according to any one of claims 11 to 13, characterized in that the lower part of the funnel-shaped cavity (44b) and the inserted tip (6) of the pin (1) form a liquid-tight seal.
  15. 15. CELL SEEDING DEVICE (40), according to any one of claims 11 to 14, characterized in that the cell seeding device (40) is made of plastic, preferably polypropylene, rubber, polystyrene, polyester and polycarbonate.
  16. 16. CELL SEEDING DEVICE (40), according to claim 11, characterized in that the funnel-shaped cavity (44) is formed as a single piece.
  17. 17. CELL SEEDING DEVICE (40), according to any one of claims 11 to 16, characterized in that the cell seeding device (40) comprises a removable reducing sleeve (470) in the upper cavity part (44a), wherein the sleeve (470) comprises a chamber with openings at both ends and the end of the sleeve near the lower cavity part (44b) has the opening (47) with a size smaller than the diameter of the pin tip (1).
  18. 18. CELL SEEDING DEVICE (40), according to any one of claims 11 to 17, characterized in that the end of the sleeve near the bottom of the cavity (44b) contains multiple micro-openings (48).
  19. 19. CELL SEEDING DEVICE (40), according to claim 18, characterized in that the sleeve (470) contains a separator (49) to form multichambers within the sleeve, wherein each chamber contains at least one micro-opening (48) at the end of the sleeve near the bottom of the cavity (44b).
  20. 20. CELL SEEDING DEVICE (40), according to claim 18, characterized in that the cavity of the upper part (44a) has the same diameter as the lower part of the cavity (44b).

Description

FIELD OF THE INVENTION [0001] The present invention provides cell assay systems and their use in cell-based assays. BACKGROUND OF THE INVENTION [0002] Cell-based assays have been widely used in the fields of cell biology, medical science, pharmacology, toxicology, and so on. They are generally performed in a multi-well cell culture plate. Cells can be in the form of suspensions, 2D monolayers, or 3D organoids/spheroids depending on the assay. To meet some special needs, many new systems have been introduced. These include microfluidic cell culture devices, 3D membrane-based perfusion bioreactors, 3D cell culture pillar systems, and so on. Despite bringing some advantages, these systems present more or less drawbacks in terms of complexity, robustness, flexibility, ease of handling, and cost-effectiveness. For example, the flow channel of the microfluidic system is prone to being blocked by air bubbles, hydrogel, and cell clumps. The 3D cell culture pillar system is only suitable for preparing multispheroids from proliferating cells because it has to use hydrogel to immobilize the cells in the pillar. Non-proliferative cells such as human hepatocytes cannot form spheroids in hydrogel. The use of hydrogel also limits the applications of this pillar system, as the hydrogel can have impacts, primarily on the diffusion of large molecules and on assay results. Furthermore, immobilizing the cells to the tiny pillar can be challenging, as the hydrogel needs to solidify in a very short time to prevent evaporation of the small volume. [0003] In view of the shortcomings of these systems, the present invention introduces a mobile pin cell culture system for cell-based assays. The system is not only suitable for 2D monolayer and 3D cell culture, but is also capable of simultaneously testing different types of cells cultured in 2D monolayer, 3D structure and without 3D structure. This system is more flexible, easy to handle and robust for performing cell-based assays. DESCRIPTION OF THE INVENTION [0004] In a first aspect, the present invention relates to a pin for use in a cell-based assay device comprising: a pin body comprising a pin head and a pin tip, wherein the pin tip comprises a surface for seeding cells. [0005] In one embodiment of the pin of the present invention, the pin body comprises at least one conical segment and the pin body forms a T shape together with the pin head. [0006] In one embodiment of the pin of the present invention, the pin body and the pin tip have a cylindrical shape. [0007] In one embodiment of the pin of the present invention, the pin body and the pin tip have a non-round cross-sectional shape. [0008] In one embodiment of the pin of the present invention, the pin head is a stepped pin head comprising a groove. [0009] In one embodiment of the pin of the present invention, the surface is selected from the group consisting of a flat surface, a flat surface with a raised edge on the margin, an inwardly structured surface, a multi-groove surface, a multi-micropit surface, a multi-micropillar surface, a rough surface, a permeable membrane, a porous segment or a magnetic segment; preferably the layer comprises a cell adhesion promoting layer, more preferably a cell adhesion promoting layer selected from hydrogel. [00010] In a second aspect, the present invention relates to a pin holder comprising a plurality of openings for receiving a pin according to the present invention, wherein the pin holder comprises a ring-shaped part connected to a cylindrical part to form a T-shaped pin holder, wherein the diameter of the ring-shaped part is greater than the diameter of the cylindrical part, wherein the ring-shaped part comprises the plurality of openings for receiving the pins. [00011] In one embodiment, the pin support is made of plastic, preferably polytetrafluoroethylene, polystyrene, polyester, and polycarbonate. [00012] In one embodiment, the pin support is sized to fit into the well of a multi-well plate. [00013] In a third aspect, the present invention relates to a device for performing a cell-based assay comprising: a pin holder comprising a plurality of openings for receiving a pin according to the present invention and a base comprising a groove channel. [00014] In one embodiment of the device for performing a cell-based assay, the pin holder and the base have a combined shape and the openings for receiving a pin are arranged so that they are aligned with the groove channel of the base. [00015] In an embodiment of the device for performing a cell-based assay, the pin holder further comprises an opening for receiving a cell culture insert and the base further comprises a reservoir for receiving a liquid, wherein the reservoir is disposed within the base so that it is aligned with the opening of the pin holder for receiving a cell culture insert. [00016] In one embodiment of the device for performing a cell-based assay, the device further comprises a pump that is fluidically connected to the