In vitro preparation of tubular tissue structures by stromal cell culture on a three-dimensional framework

• US 5,863,531 A
• Filed: 06/07/1995
• Issued: 01/26/1999
• Est. Priority Date: 04/18/1986
• Status: Expired due to Term

• Current Assignee: Smith & Nephew Wound Management (Smith & Nephew PLC)
• Original Assignee: Advanced Tissue Sciences, Inc.
• Inventors: Naughton, Gail K., Naughton, Brian A.
• Primary Examiner(s): Naff, David M.
• Application Number:
  US08/487,749
• Time in Patent Office: 1,329 Days
• Field of Search: 435/174, 435/180, 435/18.2, 435/240.23, 435/395, 435/398, 424/93.7, 424/423
• US Class Current: 424/93.7
• 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: A61F 2/0063 Implantable repair or support meshes, e.g. hernia meshes
• CPC Class Codes: A61F 2/04 Hollow or tubular parts of organs, e.g. bladders, tracheae, bronchi or bile ducts A61F2/18, A61F2/20 take precedence; devices, other than stent-grafts, providing patency to, or preventing collapsing of, tubular structures of the body other than stent-grafts, e.g. stents A61F2/82; instruments specially adapted for placement or removal of stents or stent-grafts A61F2/95
• CPC Class Codes: A61F 2/06 Blood vessels
• CPC Class Codes: A61F 2/08 Muscles; Tendons; Ligaments sutures A61B17/04
• 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 27/3629 Intestinal tissue, e.g. small intestinal submucosa
• CPC Class Codes: A61L 27/3637 characterised by the origin of the biological material other than human or animal, e.g. plant extracts, algae
• CPC Class Codes: A61L 27/3641 characterised by the site of application in the body materials for artificial blood vessels A61L27/507; materials for use in artificial skin A61L27/60
• CPC Class Codes: A61L 27/3804 characterised by specific cells or progenitors thereof, e.g. fibroblasts, connective tissue cells, kidney cells
• CPC Class Codes: A61L 27/3839 characterised by the site of application in the body materials for artificial blood vessels A61L27/507; materials for use in artificial skin A61L27/60
• CPC Class Codes: A61L 27/386 Ligaments, tendons
• CPC Class Codes: A61L 27/3886 comprising two or more cell types
• 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 together with transferrin C12N2500/25; Insulin-like growth factors C12N2501/105
• CPC Class Codes: C12N 2501/335 : Glucagon; Glucagon-like peptide [GLP]; Exendin
• CPC Class Codes: C12N 2501/39 : Steroid hormones
• CPC Class Codes: C12N 2502/094 : keratinocytes
• CPC Class Codes: C12N 2502/1323 : Adult fibroblasts
• CPC Class Codes: C12N 2502/28 : Vascular endothelial cells
• 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 thermoreactive polymers, e.g. PNIPAm, C12N2539/10
• 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 5/0062 : General methods for three-dimensional culture
• CPC Class Codes: C12N 5/066 : Tenocytes; Tendons, Ligaments
• CPC Class Codes: C12N 5/0671 : Three-dimensional culture, tissue culture or organ culture; Encapsulated cells
• CPC Class Codes: C12N 5/0676 : Pancreatic cells
• CPC Class Codes: C12N 5/0677 : Three-dimensional culture, tissue culture or organ culture; Encapsulated cells
• CPC Class Codes: C12N 5/0684 : Cells of the urinary tract or kidneys
• CPC Class Codes: C12N 5/069 : Vascular Endothelial cells
• CPC Class Codes: C12N 5/0691 : Vascular smooth muscle cells; 3D culture thereof, e.g. models of blood vessels
• CPC Class Codes: C12N 5/0697 : Artificial constructs associating cells of different lineages, e.g. tissue equivalents blood vessels C12N5/0691
• CPC Class Codes: C12N 5/0698 : Skin equivalents
• CPC Class Codes: G01N 2500/00 : Screening for compounds of potential therapeutic value
• CPC Class Codes: G01N 33/5088 : of vertebrates

Abstract

A stromal cell-based three-dimensional cell culture system is provided which can be used to culture a variety of different cells and tissues in vitro for prolonged periods of time. The stromal cells along with connective tissue proteins naturally secreted by the stromal cells attach to and substantially envelope a framework composed of a biocompatible non-living material formed into a three-dimensional structure having interstitial spaces bridged by the stromal cells. Living stromal tissue so formed provides support, growth factors, and regulatory factors necessary to sustain long-term active proliferation of cells in culture and/or cultures implanted in vivo. When grown in this three-dimensional system, the proliferating cells mature and segregate properly to form components of adult tissues analogous to counterparts in vivo, which can be utilized in the body as a corrective tissue. The three-dimensional cultures can be used to form tubular tissue structures, like those of the gastrointestinal and genitourinary tracts, as well as blood vessels; tissues for hernia repair and/or tendons and ligaments.

Claims

1. A tubular 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 three-dimensional tubular framework composed of a biocompatible, non-living material having interstitial spaces bridged by the stromal cells.

2. The tubular living stromal tissue of claim 1 in which the stromal cells are fibroblasts.

3. The tubular living stromal tissue of claim 1 in which the stromal cells are human dermal fibroblasts.

4. The tubular living stromal tissue of claim 1 in which the stromal cells are fibroblasts, pericytes, macrophages, monocytes, plasma cells, mast cells, adipocytes, umbilical cord cells, bone marrow cells from umbilical cord blood, smooth muscle cells, mucosal cells from genitourinary tract, mucosal cells from the nasopharyngeal lining, endothelial cells or a combination thereof.

5. The tubular living stromal tissue of claim 1 in which the framework is composed of a biodegradable material.

6. The tubular living, stromal tissue of claim 5 in which the biodegradable material is polyglycolic acid, cotton, cat gut sutures, cellulose, gelatin, collagen or polyhydroxyalkanoates.

7. The tubular living stromal tissue of claim 1 in which the framework is composed of a non-biodegradable material.

8. The tubular living stromal tissue of claim 7 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.

9. The tubular living stromal tissue of claim 1, 2, 3, 4, 5, 6, 7 or 8 in which the framework is a mesh.

10. A method for implantation of a tubular living stromal tissue, comprising implanting the tubular living stromal tissue of claim 1.

11. A method for preparing a tubular living stromal tissue in vitro, comprising culturing stromal cells previously inoculated onto a three-dimensional tubular framework composed of a biocompatible, non-living material in a culture medium, so that the stromal cells and connective tissue proteins secreted by the stromal cells attach to and substantially envelope the framework to produce a three-dimensional tubular structure having interstitial spaces bridged by the stromal cells.

12. The method of claim 11 in which the stromal cells are fibroblasts.

13. The method of claim 11 in which the stromal cells are human dermal fibroblasts.

14. The method of claim 11 in which the stromal cells are fibroblasts, pericytes, macrophages, monocytes, plasma cells, mast cells, adipocytes, umbilical cord cells, bone marrow cells from umbilical cord blood, smooth muscle cells, mucosal cells from the gastrointestinal tract, mucosal cells from genitourinary tract, mucosal cells from the nasopharyngeal lining, endothelial cells or a combination thereof.

15. The method of claim 11 in which the framework is composed of a biodegradable material.

16. The method of claim 15 in which the biodegradable material is polyglycolic acid, cotton, cat gut sutures, cellulose, gelatin, collagen or polyhydroxyalkanoates.

17. The method of claim 11 in which the framework is composed of a non-biodegradable material.

18. The method of claim 17 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.

19. The method of claim 11, 12, 13, 14, 15, 16, 17, or 18 in which the framework is a mesh.

20. The method of claim 11 further comprising culturing parenchymal cells inoculated onto the living stromal tissue.

21. The method of claim 20 in which the parenchymal cells comprise smooth muscle cells, endothelial cells, mucosal cells from gastrointestinal tract, mucosal cells from genitourinary tract, mucosal cells from the nasopharyngeal lining, or endothelial cells, fibroblasts, umbilical cord cells, or bone marrow cells from umbilical cord blood.

22. The method of claim 11 in which the culture medium is kept under static conditions.

23. The method of claim 11 in which the culture medium is kept in dynamic state by recirculating the culture medium.

24. A three-dimensional tubular blood vessel tissue, comprising endothelial cells cultured on a tubular living stromal tissue prepared in vitro, which stromal tissue comprises stromal cells and connective tissue proteins naturally secreted by the stromal cells attached to and substantially enveloping a three-dimensional tubular framework composed of a biocompatible, non-living material having interstitial spaces bridged by the stromal cells.

25. A three-dimensional tubular gastrointestinal epithelial tissue, comprising gastrointestinal mucosal epithelial cells cultured on a tubular living stromal tissue prepared in vitro, which stromal tissue comprises stromal cells and connective tissue proteins naturally secreted by the stromal cells attached to and substantially enveloping a three-dimensional tubular framework composed of a biocompatible, non-living material having interstitial spaces bridged by the stromal cells.

26. A three-dimensional tubular genitourinary epithelial tissue, comprising genitourinary epithelial cells cultured on a tubular living stromal tissue prepared in vitro, which stromal tissue comprises stromal cells and connective tissue proteins naturally secreted by the stromal cells attached to and substantially enveloping a three-dimensional tubular framework composed of a biocompatible, non-living material having interstitial spaces bridged by the stromal cells.

27. A method for preparing a tubular living stromal tissue in vitro, comprising culturing stromal cells previously inoculated onto a three-dimensional framework composed of a biocompatible, non-living material in a culture medium, so that the stromal cells and connective tissue proteins secreted by the stromal cells attach to and substantially envelope the framework to produce a three-dimensional structure having interstitial spaces bridged by the stromal cells;

and rolling the three-dimensional structure into a tubular living stromal tissue.