Spheroid cell culture well article and methods thereof

Patent metadata

Field	Value
Publication number	US-9790465-B2
Application number	US-201314087906-A
Country	US
Kind code	B2
Filing date	Nov 22, 2013
Priority date	Apr 30, 2013
Publication date	Oct 17, 2017
Grant date	Oct 17, 2017

Abstract

A spheroid cell culture article including:
a frame having a chamber including:
an opaque side wall surface; a top aperture; a gas-permeable, transparent bottom; and optionally a chamber annex surface and second bottom,

and at least a portion of the transparent bottom includes at least one concave arcuate surface, is disclosed. Methods of making and using the article are also disclosed.

First claim

What is claimed is:

1. A cell culture article comprising:
a chamber comprising:
an opaque side wall;
a top aperture; and
a gas-permeable, liquid impermeable bottom comprising at least one concave surface, wherein at least a portion of the bottom is transparent and wherein at least a portion of the bottom comprises a low-adhesion or no-adhesion material in or on the at least one concave surface.

2. The article of claim 1 further comprising a liner having a porous membrane situated within a portion of the chamber to divide the chamber into an upper chamber and a lower chamber.

3. The article of claim 1 further comprising a chamber annex for receiving a pipette tip for aspiration, the chamber annex comprises a surface adjacent to and in fluid communication with the chamber, the chamber annex having a second bottom spaced away and at an elevation above the gas-permeable, transparent bottom, wherein the second bottom deflects fluid dispensed from a pipette away from the transparent bottom.

4. The article of claim 3 further comprising a liner having a porous membrane situated within a portion of the chamber to form an upper chamber and a lower chamber the porous membrane providing separation of a second cellular material in the upper chamber, from a first cellular material in the lower chamber .

5. The article of claim 1 wherein the article comprises from 1 to about 2,000 of said chambers, wherein each chamber is physically separated from any other chamber.

6. The article of claim 1 wherein the at least one concave surface comprises a plurality of concave surfaces within the same chamber.

7. The article of claim 1 wherein the at least one concave surface comprises a hemi-spherical surface, a conical surface having a taper of 30 to about 60 degrees from the opaque side walls to the bottom, or a combination thereof.

8. The article of claim 1 wherein the opaque side wall surface comprises a vertical cylinder, a portion of a vertical conic of decreasing diameter from the chamber's top to bottom surface, a vertical square shaft having a conical transition to the at least one concave bottom surface, or a combination thereof.

9. The article of claim 1 wherein the at least one concave surface comprises a hemisphere, or a portion thereof, having a diameter of from about 250 to about 5,000 microns.

10. The article of claim 1 , further comprising culture media and a spheroid in said chamber.

11. The article of claim 1 , wherein said opaque side wall is adjacent to said transparent portion of said bottom.

12. The article of claim 1 , further comprising one or more auxiliary chambers in fluid communication with said chamber.

13. A perfusion plate apparatus comprising:
at least one cell culture article of claim 1 ;
a media source well in fluid communication with at least one chamber of the cell culture article that controllably provides a source fresh media to the at least one chamber; and a waste well in fluid communication with the at least one chamber of the cell culture article that controllably receives waste media from the at least one chamber.

14. The apparatus of claim 13 further comprising a porous liner situated within a portion of the at least one chamber to form an upper chamber and a lower chamber, the porous liner providing separation of a second cellular material in the upper portion of the chamber, from a first cellular material in the lower chamber.

15. The apparatus of claim 13 further comprising: a perfusion plug between the source well and the at least one chamber, a perfusion plug between the at least one chamber and the waste well, or both.

16. The apparatus of claim 13 wherein the at least one concave surface comprises a plurality of adjacent concave arcuate surfaces.

17. A method of making the article of claim 1 comprising:
combining the gas-permeable, transparent, bottom portion and an opaque side wall surface portion to form the article.

18. The method of claim 17 wherein combining is accomplished with: injection molding, thermal reforming, post forming, distortion printing, over molding, or combinations thereof.

19. The method of claim 17 wherein the gas-permeable, transparent, bottom portion includes a distortion collar integral with or attached to the gas permeable transparent bottom portion to prevent opaque side wall material from entering the arcuate bottom portion.

20. The method of claim 17 further comprising inserting a porous membrane in at least one of the chambers of the resulting article.

21. A method of culturing spheroids comprising:
charging the article of claim 1 with culture media; and
adding spheroid forming cells to the culture media.

22. The method of claim 21 further comprising optically examining the cultured spheroid forming cells through the transparent bottom.

Assignees

• Corning Inc

Inventors

• Bennett Scott Matthew
• Douglass Brian Robb
• Gagnon Jr Paul Ernest
• Martin Gregory Roger
• Szlosek Paul Michael
• Tanner Allison Jean

Classifications

• C12M23/24

  Gas permeable parts · CPC title

• C12M23/02 Primary

  Form or structure of the vessel (large containers B65D88/00) · CPC title

• C12M29/04

  Filters; Permeable or porous membranes or plates, e.g. dialysis · CPC title

• C12M23/12

  Well or multiwell plates (C12M25/04 takes precedence) · CPC title

• C12M25/04

  in combination with well or multiwell plates, i.e. culture inserts · CPC title