Classifications

• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
  • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
  • A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
  • A61L27/507—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials for artificial blood vessels
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
  • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
  • A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
  • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
  • A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
  • A61L27/60—Materials for use in artificial skin
• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
  • C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
  • C12N5/0068—General culture methods using substrates
  • C12N5/0075—General culture methods using substrates using microcarriers
• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
  • C12N2533/00—Supports or coatings for cell culture, characterised by material
  • C12N2533/20—Small organic molecules
• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
  • C12N2533/00—Supports or coatings for cell culture, characterised by material
  • C12N2533/70—Polysaccharides
• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
  • C12N2533/00—Supports or coatings for cell culture, characterised by material
  • C12N2533/70—Polysaccharides
  • C12N2533/76—Agarose, agar-agar

Definitions

• This invention relates to a surface for the attachment and growth of cells, processes for preparing said surface, and methods of promoting cell adhesion and growth on said surface.
• the proliferation of many types of cells such as mammalian cells only takes place if there is a compatible surface on which the cells will adhere and spread.
• the compatibility of a culture surface is believed to be related to its ability to adsorb specific adhesive proteins from the serum component of the growth medium.
• a variety of chemically unrelated substances, including proteins, plastics, polysaccharides, metals and ceramics will support cell growth in culture, and in most cases it seems that cell attachment and growth is mediated by the adhesive protein, fibronectin, being adsorbed onto the substrate.
• Another serum protein, vitronectin or serum spreading factor has also been proposed as a major promotor of cell adhesion in routine tissue culture.
• adhesive proteins such as fibronectin, fibronectin-fragments and vitronectin are recognised as being relatively unstable when bound to substrate, due to their molecular complexity and structure-function specificity.
• indoles selected from the group containing serotonin, tryptamine, L-tryptophan and analogues thereof, adsorb from serum containing culture media adhesive proteins such as vitronectin, fibronectin or fibronectin fragments having a strong affinity for a variety of cells.
• serum containing culture media adhesive proteins such as vitronectin, fibronectin or fibronectin fragments having a strong affinity for a variety of cells.
• serotonin (5-hydroxytryptamine) was believed to interact strongly with glycoconjugates only.
• serotonin is immobilized by covalent coupling to agarose beads, it has a high affinity for glycoproteins and glycopeptides, and acts as an adhesive surface for growth of mammalian cells.
• the present inventors have now found that serotonin and other indole analogues adsorb serum proteins to provide a functionally stable complex, which complex has a strong affinity for a variety of cells. This property of the indole group to adsorb adhesive serum proteins was not recognised prior to the work of the present inventors.
• a surface for the growth of cells comprising an adhesive serum protein adsorbed to an indole selected from the group containing serotonin, tryptamine, L-tryptophan and analogues thereof, said indole-protein complex being immobilized to an appropriate substrate.
• the surface may optionally include adherent cells, which effectively act as a starter culture to facilitate the further growth of cells on such surfaces.
• the invention also provides a process for preparing a surface for the adhesion and growth of cells, said process comprising immobilizing an indole selected from the group containing serotonin, tryptamine, L-tryptophan and analogues thereof to an appropriate substrate and exposing adhesive serum proteins thereto, whereupon said indole adsorbs said proteins to form an immobilized indole-protein complex for the growth of cells.
• the surface so prepared may optionally be exposed to a predetermined concentration of cells to provide a starter culture for the subsequent adhesion and growth of cells.
• a method of promoting cell adhesion and growth on a substrate comprising exposing cells to a surface comprising an adhesive serum protein adsorbed to an indole selected from the group containing serotonin, tryptamine, L-tryptophan and analogues thereof, said indole-protein complex being immobilized to an appropriate substrate.
• the invention also provides a method of promoting cell adhesion and growth on a surface, which method comprises a) providing a surface comprising an indole selected from the group containing serotonin, tryptamine, L-tryptophan and analogues thereof, immobilized to an appropriate substrate, and, b) exposing said surface to a medium containing . biological cells and adhesive proteins.
• a surface comprising an indole selected from the group containing serotonin, tryptamine, L-tryptophan and analogues thereof, immobilized to an appropriate substrate
• a medium containing . biological cells and adhesive proteins e.g., the surfaces and methods of the invention may be used both in vitro and in vivo.
• the surfaces and methods of the invention are likely to find application wherever surfaces need to be populated by adherent cells.
• the possibility of derivitizing the indoles of the invention onto agarose and other substrates opens the way to applying the invention to provide bioimplants such as vascular
• a human tissue implant incorporating the novel surfaces of the invention.
• the substrate used in producing the surface may be any polymer, ceramic or metal considered appropriate by persons skilled in the art.
• these polymers include agarose, collagen, polyvinyl alcohol, polyester, polytetrafluoroethylene, polyurethane, silicone rubber and polyhydroxyethyl methacrylate, and any other polymers having free -OH groups or free -SH groups which present sites for covalent coupling.
• ceramic substrates include alumina, partially stabilized zirconium, carbon and silicon nitride.
• metallic substrates include platinum, titanium and iridium.
• the substrate is not to be restricted to the examples specified herein, and that other substrates may be used appropriately. Immobilization of indole onto substrate can occur by covalent coupling or radiation grafting techniques (including UV radiation grafting) commonly practiced in the art. Covalent coupling is preferably carried out in the presence of cyanogen bromide.
• the adhesive serum proteins most commonly used are fibronectin, vitronectin, and fibronectin-fragments. DESCRIPTION OF PREFERRED EMBODIMENTS
• Cells were seeded in plastic tissue culture dishes or seeded in spinner flasks at 10 5 cells/ml with modified agarose beads at 1 g/100 ml.
• BAE Bovine aortal endothelial cells
• BSM Bovine smooth muscle cells
• BHK Baby hamster kidney cells

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• Health & Medical Sciences (AREA)
• Life Sciences & Earth Sciences (AREA)
• Chemical & Material Sciences (AREA)
• General Health & Medical Sciences (AREA)
• Engineering & Computer Science (AREA)
• Public Health (AREA)
• Biomedical Technology (AREA)
• Dermatology (AREA)
• Medicinal Chemistry (AREA)
• Oral & Maxillofacial Surgery (AREA)
• Transplantation (AREA)
• Epidemiology (AREA)
• Animal Behavior & Ethology (AREA)
• Veterinary Medicine (AREA)
• Bioinformatics & Cheminformatics (AREA)
• Wood Science & Technology (AREA)
• Biotechnology (AREA)
• Organic Chemistry (AREA)
• Zoology (AREA)
• Genetics & Genomics (AREA)
• Chemical Kinetics & Catalysis (AREA)
• Microbiology (AREA)
• Botany (AREA)
• Biochemistry (AREA)
• General Engineering & Computer Science (AREA)
• Cell Biology (AREA)
• Vascular Medicine (AREA)
• Micro-Organisms Or Cultivation Processes Thereof (AREA)
• Apparatus Associated With Microorganisms And Enzymes (AREA)
• Immobilizing And Processing Of Enzymes And Microorganisms (AREA)

Applications Claiming Priority (2)

Publications (2)

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ID=3771758

Family Applications (1)

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Citations (2)

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• 1987
  • 1987-08-17 JP JP62505089A patent/JPH01500641A/ja active Pending
  • 1987-08-17 WO PCT/AU1987/000266 patent/WO1988001279A1/en not_active Application Discontinuation
  • 1987-08-17 EP EP19870905567 patent/EP0277993A4/en not_active Withdrawn

Patent Citations (2)

Non-Patent Citations (1)

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