Array of micromolded structures for sorting adherent cells

Patent metadata

Field	Value
Publication number	US-11046926-B2
Application number	US-201815945294-A
Country	US
Kind code	B2
Filing date	Apr 4, 2018
Priority date	Feb 16, 2010
Publication date	Jun 29, 2021
Grant date	Jun 29, 2021

Abstract

An apparatus for collecting or culturing cells or cell colonies includes: a common substrate formed from a flexible resilient polymeric material and having a plurality of wells formed therein; and a plurality of rigid cell carriers releasably connected to said common substrate, with said carriers arranged in the form of an array, and with each of the carriers resiliently received in one of the wells. A method of collecting or culturing cells or cell colonies with such an apparatus is carried out by depositing a liquid media carrying cells on the apparatus so that said cells settle on or adhere to said the carriers; and then (c) releasing at least one selected carrier having said cells thereon by gradual application of release energy to each carrier from the cavity in which it is received (e.g., by pushing with a probe).

First claim

That which is claimed is:

1. An apparatus for collecting or culturing cells or cell colonies, said apparatus comprising:
a substrate formed from a flexible resilient polymeric material and having a first surface and an opposed second surface and a plurality of wells formed in the first surface in the form of an array, the wells in the substrate defined by side walls,
a housing for supporting the substrate, and
a plurality of rigid cell carriers, each carrier disposed in one of said wells such that the carrier engages the side walls of the well and is resiliently held in one of said wells, the carriers configured to release from said substrate upon mechanical distortion of said substrate.

2. The apparatus of claim 1 , wherein said carriers are transparent or semitransparent.

3. The apparatus of claim 1 wherein:
said wells in said substrate are separated by walls, said walls have an average width of at least 2 micrometers, up to 1000 micrometers;
and said walls have an average height of at least 2 micrometers, up to 1000 micrometers;
said wells in said substrate have floors, and said floors have an average thickness of from 2 to 500 micrometers;
said substrate has a top surface and said carriers have a top surface, and said carrier top surfaces are positioned at or below said substrate top surface;
said carriers have heights of at least 2 micrometers, up to 500 micrometers; and
said carriers have maximum widths of at least 5 micrometers, up to 1000 micrometers.

4. The apparatus of claim 1 , wherein said substrate has a top surface and said carriers have a top surface, and wherein said carrier top surfaces are positioned at or below said substrate top surface.

5. The apparatus of claim 1 , wherein said carriers are produced by the process of casting said carriers in said wells.

6. The apparatus of claim 1 , wherein said carriers have a concave top surface portion.

7. The apparatus of claim 6 , wherein said concave top surface portion is formed by meniscus coating of the side walls of said wells during the process of casting said carriers in said wells.

8. The apparatus of claim 1 , wherein said carriers are coated with a biologically active molecule on at least the top surface thereof.

9. The apparatus of claim 1 , wherein said carriers are magnetic or ferromagnetic.

10. The apparatus of claim 1 , wherein said carriers comprise a plurality of layers.

11. The apparatus of claim 1 , wherein said carriers comprise polystyrene.

12. The apparatus of claim 1 , wherein said carriers comprise an anionic transparent magnetic polystyrene.

13. The apparatus of claim 1 , wherein said carriers comprise:
a rigid lower layer; and
a cell-growth compatible upper layer.

14. The apparatus of claim 1 , wherein said carriers comprise a hydrogel upper layer, said hydrogel containing live feeder cells.

15. The apparatus for collecting or culturing cells or cell colonies as recited in claim 1 , wherein the polymeric material is selected from silicones, polyurethane, styrene-butadiene copolymer, polyolefin and polydiene elastomers, thermoplastic elastomers or combinations thereof.

16. The apparatus for collecting or culturing cells or cell colonies as recited in claim 1 , wherein the array is uniform or non-uniform.

17. The apparatus for collecting or culturing cells or cell colonies as recited in claim 1 , wherein the array is arranged in an interdigitated pattern.

18. The apparatus for collecting or culturing cells or cell colonies as recited in claim 1 , wherein the housing comprises a cassette formed from polycarbonate or glass.

19. The apparatus for collecting or culturing cells or cell colonies as recited in claim 18 , wherein the cassette includes a scaffold.

20. The apparatus for collecting or culturing cells or cell colonies as recited in claim 1 , wherein the shape of the cell carriers is cylindrical, elliptical, triangular, rectangular, square, hexagonal, pentagonal, octagonal or combinations thereof.

Assignees

• Univ North Carolina Chapel Hill

Inventors

• Allbritton Nancy
• Sims Christopher
• Wang Yuli
• Shah Pavak Kirit

Classifications

• C12N2509/10

  Mechanical dissociation · CPC title

• C12N5/0068

  General culture methods using substrates (for specific animal cell type C12N5/06) · CPC title

• C12N1/02

  Separating microorganisms from their culture media · CPC title

• B01J2219/00743

  Cells · CPC title

• B01J2219/00452

  Means for the recovery of reactants or products · CPC title