Three-dimensional cell and tissue culture apparatus

• US 5,160,490 A
• Filed: 02/21/1991
• Issued: 11/03/1992
• Est. Priority Date: 04/18/1986
• Status: Expired due to Term

• Current Assignee: RegeneMed, Inc.
• Original Assignee: Advanced Tissue Sciences, Inc.
• Inventors: Naughton, Gail K., Naughton, Brian A.
• Primary Examiner(s): Robinson, Douglas W.
• Assistant Examiner(s): WILLIAMS, JANE
• Application Number:
  US07/659,220
• Time in Patent Office: 621 Days
• Field of Search: 435/297, 435/300, 435/301, 435/32, 435/2, 435/4, 435/240.2, 435/240.243, 435/240.23, 435/240.21, 435/240.1, 435/284
• US Class Current: 435/287.1
• CPC Class Codes: A61B 2010/0258 Marrow samples
• CPC Class Codes: A61B 90/00 Instruments, implements or accessories specially adapted for surgery or diagnosis and not covered by any of the groups A61B1/00 - A61B50/00, e.g. for luxation treatment or for protecting wound edges protective face masks A41D13/11; surgeon's or patient's gowns or dresses A41D13/12; devices for carrying-off, for treatment of, or for carrying-over, body liquids A61M1/00
• CPC Class Codes: A61K 2035/124 the cells being hematopoietic, bone marrow derived or blood cells
• CPC Class Codes: A61K 35/12 Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells vaccines or medicinal preparations containing antigens or antibodies A61K39/00
• CPC Class Codes: A61K 35/28 Bone marrow; Haematopoietic stem cells; Mesenchymal stem cells of any origin, e.g. adipose-derived stem cells
• CPC Class Codes: A61L 27/38 containing added animal cells organs or tissue containing native cells A61L27/36
• CPC Class Codes: A61L 27/60 Materials for use in artificial skin
• CPC Class Codes: C12N 2500/34 Sugars
• CPC Class Codes: C12N 2501/11 Epidermal growth factor [EGF]
• CPC Class Codes: C12N 2501/33 Insulin
• CPC Class Codes: C12N 2501/335 Glucagon; Glucagon-like peptide [GLP]; Exendin
• CPC Class Codes: C12N 2501/39 Steroid hormones
• CPC Class Codes: C12N 2502/091 melanocytes
• CPC Class Codes: C12N 2502/094 keratinocytes
• CPC Class Codes: C12N 2502/1323 Adult fibroblasts
• CPC Class Codes: C12N 2503/04 Screening or testing on artificial tissues
• CPC Class Codes: C12N 2510/00 Genetically modified cells
• CPC Class Codes: C12N 2533/30 Synthetic polymers
• CPC Class Codes: C12N 2533/40 Polyhydroxyacids, e.g. polymers of glycolic or lactic acid (PGA, PLA, PLGA); Bioresorbable polymers
• CPC Class Codes: C12N 2533/70 Polysaccharides
• CPC Class Codes: C12N 5/0698 : Skin equivalents

Abstract

The present invention relates to a three-dimensional cell culture system which can be used to culture a variety of different cells and tissues in vitro for prolonged periods of time. In accordance with the invention, cells derived from a desired tissue are inoculated and grown on a pre-established stromal support matrix. The stromal support matrix comprises stromal cells, such as fibroblasts actively growing on a three-dimensional matrix. Stromal cells may also include other cells found in loose connective tissue such as endothelial cells, macrophages/monocytes, adipocytes, pericytes, reticular cells found in bone marrow stroma, etc. The stromal matrix provides the support, growth factors, and regulatory factors necessary to sustain long-term active proliferation of cells in culture. When grown in this three-dimensional system, the proliferating cells mature and segregate properly to form components of adult tissues analogous to counterparts found in vivo.

Claims

1. A cytological testing apparatus, comprising a three dimensional cell culture positioned in a container to which a test substance can be added, in which the three dimensional cell culture comprises parenchymal cells cultured on a living stromal tissue prepared in vitro comprising stromal cells and connective tissue proteins naturally secreted by the stromal cells attached to and substantially enveloping a framework composed of a biocompatible, non-living material formed to a three dimensional structure having interstitial spaces bridged by the stromal cells.

2. The cytological testing apparatus of claim 1 in which the stromal cells are fibroblasts.

3. The cytological testing apparatus of claim 1 in which the stromal cells are a combination of fibroblasts and anyone or more of the following stromal cell types:

endothelial cells, pericytes, macrophages, monocytes, leukocytes, plasma cells, mast cells or adipocytes.

4. The cytological testing apparatus of claim 1 in which the framework is composed of a biodegradable material.

5. The cytological testing apparatus of claim 4 in which the biodegradable material is cotton, polyglycolic acid, cat gut sutures, cellulose, gelatin or dextran.

6. The cytological testing apparatus of claim 1 in which the framework is composed of a non-biodegradable material.

7. The cytological testing apparatus of claim 6 in which the non-biodegradable material is a polyamide, a polyester, a polystyrene, a polypropylene, a polyacrylate, a polyvinyl, a polycarbonate, a polytetrafluorethylene, or a nitrocellulose compound.

8. The cytological testing apparatus of claim 4, 5, 6, or 7 in which the framework is precoated with collagen.

9. The cytological testing apparatus of claim 1, 2, 3, 4, 5, 6, or 7 in which the framework is a mesh.

10. The cytological testing apparatus of claim 8 in which the framework is a mesh.

11. The cytological testing apparatus of claim 1, 2, 3, 4, 5, 6, or 7 in which the container is a multi-welled container.

12. The cytological testing apparatus of claim 8 in which the container is a multi-welled container.

13. The cytological testing apparatus of claim 9 in which the container is a multi-welled container

14. The cytological testing apparatus of claim 1 in which the container is a multi-welled container.

15. The cytological testing apparatus of claim 1 in which the parachymal cells are hematopoietic cells.

16. The cytological testing apparatus of claim 1 in which the parachymal cells are melancocytes and keratincocytes.

17. The cytological testing apparatus of claim 1 in which the parenchymal cells are hepatocytes.

18. The cytological testing apparatus of claim 1 in which the parenchymal cells are pancreatic acinar cells.

19. The cytological testing apparatus of claim 1 in which the parenchymal cells are kidney cells.