Method for altering the differentiation of anchorage dependent cells on an electrically conducting polymer

• US 5,843,741 A
• Filed: 08/01/1994
• Issued: 12/01/1998
• Est. Priority Date: 08/01/1994
• Status: Expired due to Fees

• Current Assignee: Children's Medical Center Corporation, Massachusetts Institute of Technology
• Original Assignee: Children's Medical Center Corporation, Massachusetts Institute of Technology
• Inventors: Wong, Joyce Y., Langer, Robert S., Ingber, Donald E.
• Primary Examiner(s): Weber, Jon P.
• Application Number:
  US08/283,402
• Time in Patent Office: 1,583 Days
• Field of Search: 435/173.8
• US Class Current: 435/173.8
• CPC Class Codes: C12N 13/00 Treatment of microorganisms or enzymes with electrical or wave energy, e.g. magnetism, sonic waves
• CPC Class Codes: C12N 2533/30 Synthetic polymers thermoreactive polymers, e.g. PNIPAm, C12N2539/10
• CPC Class Codes: C12N 5/0068 General culture methods using substrates for specific animal cell type C12N5/06

Abstract

Described is a method and cell culture system for altering the proliferation, differentiation, or function of anchorage dependent cells which includes associating the cells with a surface formed of an electrically conducting polymer and applying an effective amount of a voltage to change the oxidation state of the polymer without damaging the cells.

Claims

1. A method for altering the differentiation of anchorage dependent cells comprising

binding cell surface receptor-specific attachment molecules of defined types and in a defined density to a surface consisting of an electrically conducting polymer,

attaching anchorage-dependent cells to the cell surface receptor-specific attachment molecules on the electrically conducting polymer surface, and

applying a voltage in an effective amount to change the oxidation state of the electrically conducting polymer thereby to alter the differentiation as a function of the type and density of the attachment molecules and applied voltage,

wherein the anchorage dependent cells are selected from the group consisting of animal cells and plant cells.

2. The method of claim 1 wherein the electrically conductive polymer is selected from the group consisting of polyacetylene, polyaniline, polypyrrole, polythiophene, poly(phenylenesulfide), poly(phenylenevinylene), copolymers and blends thereof.

3. The method of claim 1 wherein the cells are dissociated mammalian cells.

4. The method of claim 1 wherein the polymer is coated onto a device or structure for cell culture.

5. The method of claim 1 wherein the polymer is coated onto, or used to form, a device or structure for implantation in the body.

6. The method of claim 5 wherein the device or structure is selected from the group consisting of sutures, bone screws, nails, plates, tubes, sheets, films, stents, and scaffolds for tissue engineering.

7. The method of claim 5 further comprising, following binding cell surface receptor-specific attachment molecules and prior to applying a voltage, implanting the polymeric material into a patient in need of treatment, wherein applying a voltage to the material alters healing of a wound in the body of the patient wherein the alteration is to enhance.

8. The method of claim 5 further comprising, following binding cell surface receptor-specific attachment molecules and prior to applying a voltage, implanting the polymeric material into a patient in need of treatment, wherein applying a voltage to the material alters formation of adhesions in the body of the patient, wherein the alteration is to decrease.

9. The method of claim 5 further comprising, following binding cell surface receptor-specific attachment molecules and prior to applying a voltage, implanting the polymeric material into a patient in need of treatment, wherein applying a voltage to the material alters differentiation and organ function in the body of the patient, wherein the alteration is to stimulate.

10. The method of claim 5 further comprising, following binding cell surface receptor-specific attachment molecules and prior to applying a voltage, implanting the polymeric material into a patient in need of bone repair or regeneration, wherein applying a voltage to the material alters bone repair or regeneration in the body of the patient, wherein the alteration is to enhance.

11. The method of claim 5 further comprising, following binding cell surface receptor-specific attachment molecules and prior to applying a voltage, implanting the polymeric material into a patient in need of nerve repair or regeneration, wherein applying a voltage to the material alters nerve repair or regeneration in the body of the patient, wherein the alteration is to enhance.

12. The method of claim 1 wherein the attachment molecules are selected from the group consisting of gelatin, albumin, fibronectin, laminin, carbohydrates, asialoglycoproteins, lectins, growth factors, low density lipoproteins, heparin, poly-lysine, thrombin, vitronectin, fibrinogen, peptides, and mixtures thereof.

13. The method of claim 12 wherein the peptides incorporate binding sites selected from the group consisting of R G D, L I G R K K T (SEQ ID NO:

1), and Y I G S R (SEQ ID NO;

     2).

14. The method of claim 1 wherein the attachment molecules are selected from the group consisting of thrombospondin, entactin, proteoglycans, glycosaminoglycans, and mixtures thereof.

15. The method of claim 1 wherein the conducting polymer is coated onto a device or structure formed of a biodegradable polymer.

16. A device or structure for altering the differentiation of anchorage dependent cells comprising:

an electrically conductive polymer surface;

cell surface receptor-specific attachment molecules of defined types bound to the electrically conductive polymer surface in a defined density; and

a means to apply a voltage across the electrically conductive polymer surface, wherein the means is electrically connected with the polymer,

wherein the effect of the voltage on the differentiation of the cells attached to the polymer surface is altered as a function of the type and density of the attachment molecules.

17. The device or structure of claim 16 wherein the electrically conductive polymer is selected from the group consisting of polyacetylene, polyaniline, polypyrrole, polythiophene, poly(phenylenesulfide), poly(phenylenevinylene), copolymers and blends thereof.

18. The device or structure of claim 16 wherein the attachment molecules are selected from the group consisting of gelatin, albumin, fibronectin, laminin, carbohydrates, asialoglycoproteins, lectins, growth factors, low density lipoproteins, heparin, poly-lysine, thrombin, vitronectin, fibrinogen, peptides, and mixtures thereof.

19. The device or structure of claim 18, wherein the peptides incorporate binding sites selected from the group consisting of R G D, L I G R K K T (SEQ ID NO:

1), and Y I G S R (SEQ ID NO;

     2).

20. The device or structure of claim 16, wherein the attachment molecules are selected from the group consisting of thrombospondin, entactin, proteoglycans, glycosaminoglycans, and mixtures thereof.

21. The device or structure of claim 16, wherein the polymer is coated onto a device or structure for cell culture.

22. The device or structure of claim 16, wherein the polymer and the means to induce a voltage across the polymer are connected by electrodes.

23. The device or structure of claim 16, wherein the electrically conductive polymer has had a change in its oxidative state due to an applied voltage across the polymer.

24. The device or structure of claim 16, further comprising anchorage dependent cells attached to the cell surface receptor-specific attachment molecules.

25. The device or structure of claim 24 wherein the anchorage dependent cells are dissociated mammalian cells.

26. The device or structure of claim 16 wherein the device is used for implantation, and wherein the device is selected from the group consisting of sutures, bone screws, nails, plates, tubes, sheets, films, stents, and scaffolds for tissue engineering.