Cell growth chambers and method of use thereof

• US 4,975,377 A
• Filed: 05/14/1987
• Issued: 12/04/1990
• Est. Priority Date: 05/14/1987
• Status: Expired due to Fees

• Current Assignee: Marc E. Key
• Original Assignee: Marc E. Key
• Inventors: Key, Marc E.
• Primary Examiner(s): Kepplinger, Esther L.
• Assistant Examiner(s): Scheiner, Toni R.
• Application Number:
  US07/050,510
• Time in Patent Office: 1,300 Days
• Field of Search: 422/102, 206/1, 428/35, 435/284, 435/285, 435/286, 435/287, 435/297, 435/299, 435/300, 435/301, 435/240.25, 435/240.243
• US Class Current: 435/297.5
• CPC Class Codes: C12M 23/10 Petri dish crystallising dishes B01L3/06
• CPC Class Codes: C12M 23/20 Material Coatings immunocoatings C12M25/00
• CPC Class Codes: C12M 25/14 Scaffolds; Matrices in general C12N5/0068

Abstract

Growth chambers for anchorage-independent cell growth therein are formed of a gel matrix having a surface disallowing anchorage-dependent cell growth over the full interior thereof. In a preferred form the chambers have a generally cylidnrical wall and an integral convex bottom wall forming an annular volume at the foot of the cylindrical wall which is substantially lower than the central portion to concentrate such anchorage-indpendent cells. The gel matrix is sufficiently permeable to permit passage of cell-growth nutrients and waste product solutes through said wall when the chambers are filled below the open end and submerged in a growth medium. Preferably the gel matrix is formed of 1% to 5% cross-linked polyacrylamide and from 99% to 95% water.

In a preferred method of using the growth chambers, undifferentiated tumor cells and normal cells, including fibroblasts, are cultured together. Anchorage-independent tumor cells proliferate while anchorage-dependent cells are unable to grow without attachment. The method is useful for evaluating in vitro therapeutic agents to control tumor growth, normal cell growth or microspheres and generation of immunoglobulins from lymphocyte cells.

Claims

1. A cell growth chamber for anchorage independent cell growth therein, said growth chamber having a generally continuous cylindrical vertical wall of substantially uniform thickness with the foot of the interior wall surface terminating around its circumference with the outer circumference of an integral convex bottom wall to form a single cylindrical fluid container within said chamber having a single annular volume adjacent said foot of said interior cylindrical wall at a level substantially lower than the central portion of said bottom wall for concentrating the growth of anchorage-index cells in said annular volume, said chamber and bottom walls being formed of a gel matrix having a surface disallowing attachment of anchorage-dependent cells over the full interior of said chamber, and said gel matrix being capable of retaining said uniform thickness of said chamber walls while immersed in a cell-growth medium, and having a permeability sufficient to permit passage of cell-growth nutrient and waste product solutes through said cylindrical vertical wall while so immersed.

2. A cell-growth chamber in accordance with claim 1 wherein said gel matrix contains from 85% to 99.5% water.

3. A cell-growth chamber in accordance with claim 2 wherein said gel matrix is from 1% to 5% polyacrylamide and from 99% to 95% water.

4. A cell-growth chamber in accordance with claim 2 wherein said gel matrix is from 0.5% to 15% agar and from 99.5% to 85% water.

5. A cell-growth chamber in accordance with claim 1 wherein said gel matrix is a cross-linked acrylamide and sufficient water adequate to prevent cell attachment on the surface of said gel.