METHOD AND SYSTEMS FOR USING BIOPOLYMER-BASED BEADS AND HYDROGELS

• US 20080138416A1
• Filed: 06/13/2007
• Published: 06/12/2008
• Est. Priority Date: 06/13/2006
• Status: Abandoned Application

• Current Assignee: FMC Biopolymer AS (DuPont de Nemours, Inc.)
• Original Assignee: FMC Biopolymer AS (DuPont de Nemours, Inc.)
• Inventors: Lee, Randall J., Rauh, Francis, Maciejewski, Mark
• Application Number:
  US11/762,611
• Publication Number:
  US 20080138416A1
• Time in Patent Office: Days
• US Class Current: 424/488
• CPC Class Codes: A61K 38/06 Tripeptides
• CPC Class Codes: A61L 2400/06 Flowable or injectable implant compositions
• CPC Class Codes: A61L 2430/20 for reconstruction of the heart, e.g. heart valves
• CPC Class Codes: A61L 27/20 Polysaccharides
• CPC Class Codes: A61L 27/225 Fibrin; Fibrinogen
• CPC Class Codes: A61L 27/227 Other specific proteins or polypeptides not covered by A61L27/222, A61L27/225 or A61L27/24
• CPC Class Codes: A61L 27/24 Collagen
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 9/00 Drugs for disorders of the cardiovascular system
• CPC Class Codes: C08L 5/04 Alginic acid; Derivatives thereof
• CPC Class Codes: C08L 5/08 Chitin; Chondroitin sulfate; Hyaluronic acid; Derivatives thereof

Abstract

Compositions comprising biopolymers such as alginates and cell attachment peptides are disclosed. Compositions may optionally further comprise cells. Methods for repairing or treating a tissues and organs with such compositions and systems for providing such compositions to tissues and organs, and methods for delivering desired proteins to individual with such compositions and systems for providing such compositions are also disclosed. In vitro methods of culturing cells are also disclosed.

Claims

1. A method for repairing or treating a tissue or organ comprising:

providing to a tissue or organ a therapeutically effective amount of a composition selected from the group consisting of one or more of the following;

a composition comprising alginate covalently linked to two or more different cell attachment peptides;

a composition comprising alginate mixed with two or more different cell attachment peptides;

a composition comprising alginate covalently linked to one or more cell attachment peptides and mixed with one or more different cell attachment peptides;

a composition comprising alginate covalently linked to one cell attachment peptides and ionically linked to chitosan;

a composition comprising alginate covalently linked to two or more different cell attachment peptides and ionically linked to chitosan;

a composition comprising alginate mixed with one cell attachment peptides and ionically linked to chitosan;

a composition comprising alginate mixed with two or more different cell attachment peptides and ionically linked to chitosan

a composition comprising alginate covalently linked to one cell attachment peptide and mixed with cell attachment peptides and ionically linked to chitosan;

a composition comprising alginate covalently linked to two or more different cell attachment peptide and mixed with cell attachment peptides and ionically linked to chitosan;

a composition comprising cells encapsulated in alginate covalently linked to two or more different cell attachment peptides;

a composition comprising cells encapsulated in alginate mixed with two or more different cell attachment peptides;

a composition comprising cells encapsulated in alginate covalently linked to one or more cell attachment peptides and mixed with one or more cell attachment peptides;

a composition comprising cells encapsulated in alginate covalently linked to one cell attachment peptides and ionically linked to chitosan;

a composition comprising cells encapsulated in alginate covalently linked to two or more different cell attachment peptides and ionically linked to chitosan;

a composition comprising cells encapsulated in alginate mixed with one cell attachment peptides and ionically linked to chitosan;

a composition comprising cells encapsulated in alginate mixed with two or more different cell attachment peptides and ionically linked to chitosan

a composition comprising cells encapsulated in alginate covalently linked to one cell attachment peptide and mixed with cell attachment peptides and ionically linked to chitosan; and

a composition comprising cells encapsulated in alginate covalently linked to two or more different cell attachment peptide and mixed with cell attachment peptides and ionically linked to chitosan.

2. The method of claim 1 wherein the cell attachment peptides are selected from group consisting of:

RGD, YIGSR (SEQ ID NO;

1), IKVAV (SEQ ID NO;

2), REDV (SEQ ID NO;

3), DGEA (SEQ ID NO;

4), VGVAPG (SEQ ID NO;

5), GRGDS (SEQ ID NO;

6), LDV, RGDV (SEQ ID NO;

7), PDSGR (SEQ ID NO;

8), RYVVLPR (SEQ ID NO;

9), LGTIPG (SEQ ID NO;

10), LAG, RGDS (SEQ ID NO;

11), RGDF (SEQ ID NO;

12), HHLGGALQAGDV (SEQ ID NO;

13), VTCG (SEQ ID NO;

14), SDGD (SEQ ID NO;

15), GREDVY (SEQ ID NO;

16), GRGDY (SEQ ID NO;

17), GRGDSP (SEQ ID NO;

18), VAPG (SEQ ID NO;

19), GGGGRGDSP (SEQ ID NO;

20) and GGGGRGDY (SEQ ID NO;

21) and FTLCFD (SEQ ID NO;

22).

3. The method of claim 1 wherein said composition comprises alginate beads.

4. The method of claim 3 wherein the beads have mean diameters in a range of from about 30 um to about 500 um.

5. The method of claim 1 wherein said composition comprises an alginate hydrogel.

6. The method of claim 1 wherein said composition comprises cells selected from the group consisting of:

stem cells, muscle cells, pancreatic islets, chondrocytes, hepatic cells, neural cells, renal cortex cells, vascular endothelial cells, thyroid and parathyroid cells, adrenal cells, thymic cells, ovarian cells, and cells which include recombinant genetic material provided to express desired proteins.

7. The method of claim 1 wherein said composition further comprises one or more compounds selected from the group consisting of:

EGF, VEGF, b-FGF, FGF, TGF, TGF-β and

proteoglycans.

8. The method of claim 1 wherein said composition is provided to a tissue selected from the group consisting of:

bones, muscle, cartilage, connective, neural, epithelial, vascular, urothelial and mucosal.

9. The method of claim 1 wherein said composition is provided to an organ selected from the group consisting of:

skin, sinus, liver, lungs, kidney, brain, pancreas, endocrine organs, esophagus, stomach, intestines, colon, rectum, cornea, and breast.

10. The method of claim 1 wherein said composition is provided as a cross-linked matrix.

11. The method of claim 1 wherein said composition produces a matrix in situ following application to the tissue or organ.

12. A system for repairing or treating a tissue or organ comprising a delivery device for providing a composition to a tissue or organ and a therapeutically effective amount of a composition selected from the group consisting of one or more of the following:

a composition comprising alginate covalently linked to two or more different cell attachment peptides;

a composition comprising alginate mixed with two or more different cell attachment peptides;

a composition comprising alginate covalently linked to one or more cell attachment peptides and mixed with one or more cell attachment peptides;

a composition comprising alginate covalently linked to one cell attachment peptides and ionically linked to chitosan;

a composition comprising alginate covalently linked to two or more different cell attachment peptides and ionically linked to chitosan;

a composition comprising alginate mixed with one cell attachment peptides and ionically linked to chitosan;

a composition comprising alginate mixed with two or more different cell attachment peptides and ionically linked to chitosan

a composition comprising alginate covalently linked to one cell attachment peptide and mixed with cell attachment peptides and ionically linked to chitosan;

a composition comprising alginate covalently linked to two or more different cell attachment peptide and mixed with cell attachment peptides and ionically linked to chitosan;

a composition comprising cells encapsulated in alginate covalently linked to two or more different cell attachment peptides;

a composition comprising cells encapsulated in alginate mixed with two or more different cell attachment peptides;

a composition comprising cells encapsulated in alginate covalently linked to one or more cell attachment peptides and mixed with one or more cell attachment peptides;

a composition comprising cells encapsulated in alginate covalently linked to one cell attachment peptides and ionically linked to chitosan;

a composition comprising cells encapsulated in alginate covalently linked to two or more different cell attachment peptides and ionically linked to chitosan;

a composition comprising cells encapsulated in alginate mixed with one cell attachment peptides and ionically linked to chitosan;

a composition comprising cells encapsulated in alginate mixed with two or more different cell attachment peptides and ionically linked to chitosan

a composition comprising cells encapsulated in alginate covalently linked to one cell attachment peptide and mixed with cell attachment peptides and ionically linked to chitosan; and

a composition comprising cells encapsulated in alginate covalently linked to two or more different cell attachment peptide and mixed with cell attachment peptides and ionically linked to chitosan.

13. The system of claim 12 wherein said delivery device comprises two lumens.

14. The system of claim 12 wherein said delivery device comprises a syringe.

15. The system of claim 12 wherein the cell attachment peptides are selected from group consisting of:

RGD, YIGSR (SEQ ID NO;

1), IKVAV (SEQ ID NO;

2), REDV (SEQ ID NO;

3), DGEA (SEQ ID NO;

4), VGVAPG (SEQ ID NO;

5), GRGDS (SEQ ID NO;

6), LDV, RGDV (SEQ ID NO;

7), PDSGR (SEQ ID NO;

8), RYVVLPR (SEQ ID NO;

9), LGTIPG (SEQ ID NO;

10), LAG, RGDS (SEQ ID NO;

11), RGDF (SEQ ID NO;

12), HHLGGALQAGDV (SEQ ID NO;

13), VTCG (SEQ ID NO;

14), SDGD (SEQ ID NO;

15), GREDVY (SEQ ID NO;

16), GRGDY (SEQ ID NO;

17), GRGDSP (SEQ ID NO;

18), VAPG (SEQ ID NO;

19), GGGGRGDSP (SEQ ID NO;

20) and GGGGRGDY (SEQ ID NO;

21) and FTLCFD (SEQ ID NO;

22).

16. The system of claim 12 wherein said composition comprises alginate beads.

17. The system of claim 16 wherein the beads have mean diameters in a range of from about 30 um to about 500 um.

18. The system of claim 12 wherein said composition comprises an alginate hydrogel.

19. The system of claim 12 wherein said composition comprises cells selected from the group consisting of:

stem cells, muscle cells, pancreatic islets, chondrocytes, hepatic cells, neural cells, renal cortex cells, vascular endothelial cells, thyroid and parathyroid cells, adrenal cells, thymic cells, ovarian cells, and cells which include recombinant genetic material provided to express desired proteins.

20. The system of claim 12 wherein said composition further comprises one or more compounds selected from the group consisting of:

EGF, VEGF, b-FGF, FGF, TGF, TGF-β and

proteoglycans.

21. A system for repairing or treating a tissue or organ comprising a delivery device for providing two or more components to a tissue or organ, a first component comprising a gelling ion and a therapeutically effective amount of a second component selected from the group consisting of one or more of the following:

a composition comprising non-crosslinked alginate covalently linked to two or more different cell attachment peptides, and optionally further comprising cells;

a composition comprising non-crosslinked alginate mixed with two or more different cell attachment peptides and optionally further comprising cells; and

a composition comprising non-crosslinked alginate covalently linked to one or more cell attachment peptides and mixed with one or more cell attachment peptides and optionally further comprising cells.

22. The system of claim 21 wherein said delivery device comprises a first lumen for delivery of the first component and a second lumen for delivery of the second component.

23. The system of claim 21 wherein said delivery device comprises a syringe having two chambers, a first chamber for containing the first component and the second chamber for containing the second component.

24. The system of claim 21 wherein the cell attachment peptides are selected from group consisting of one or more of the following peptides:

RGD, YIGSR (SEQ ID NO;

1), IKVAV (SEQ ID NO;

2), REDV (SEQ ID NO;

3), DGEA (SEQ ID NO;

4), VGVAPG (SEQ ID NO;

5), GRGDS (SEQ ID NO;

6), LDV, RGDV (SEQ ID NO;

7), PDSGR (SEQ ID NO;

8), RYVVLPR (SEQ ID NO;

9), LGTIPG (SEQ ID NO;

10), LAG, RGDS (SEQ ID NO;

11), RGDF (SEQ ID NO;

12), HHLGGALQAGDV (SEQ ID NO;

13), VTCG (SEQ ID NO;

14), SDGD (SEQ ID NO;

15), GREDVY (SEQ ID NO;

16), GRGDY (SEQ ID NO;

17), GRGDSP (SEQ ID NO;

18), VAPG (SEQ ID NO;

19), GGGGRGDSP (SEQ ID NO;

20) and GGGGRGDY (SEQ ID NO;

21) and FTLCFD (SEQ ID NO;

22).

25. The system of claim 21 wherein said cells are selected from the group consisting of:

stem cells, muscle cells, pancreatic islets, chondrocytes, hepatic cells, neural cells, renal cortex cells, vascular endothelial cells, thyroid and parathyroid cells, adrenal cells, thymic cells, ovarian cells, and cells which include recombinant genetic material provided to express desired proteins.

26. A system for repairing or treating a tissue or organ comprising a delivery device for providing two or more components to a tissue or organ, a first component comprising a mixture of gelling ions and one or more cell attachment peptides;

and a therapeutically effective amount of a second component selected from the group consisting of one or more of the following;

a composition comprising non-crosslinked alginate, and optionally further comprising cells;

a composition comprising non-crosslinked alginate covalently linked to one or more cell attachment peptides, and optionally further comprising cells;

a composition comprising non-crosslinked alginate mixed with one or more cell attachment peptides and optionally further comprising cells; and

a composition comprising non-crosslinked alginate covalently linked to one or more cell attachment peptides and mixed with one or more cell attachment peptides and optionally further comprising cells;

wherein said first component and said second component collectively comprise two or more different cell attachment peptides.

27. The system of claim 26 wherein said delivery device comprises a first lumen for delivery of the first component and a second lumen for delivery of the second component.

28. The system of claim 26 wherein said delivery device comprises a syringe having two chambers, a first chamber for containing the first component and the second chamber for containing the second component.

29. The system of claim 26 wherein the cell attachment peptides are selected from group consisting of one or more of the following peptides:

RGD, YIGSR (SEQ ID NO;

1), IKVAV (SEQ ID NO;

2), REDV (SEQ ID NO;

3), DGEA (SEQ ID NO;

4), VGVAPG (SEQ ID NO;

5), GRGDS (SEQ ID NO;

6), LDV, RGDV (SEQ ID NO;

7), PDSGR (SEQ ID NO;

8), RYVVLPR (SEQ ID NO;

9), LGTIPG (SEQ ID NO;

10), LAG, RGDS (SEQ ID NO;

11), RGDF (SEQ ID NO;

12), HHLGGALQAGDV (SEQ ID NO;

13), VTCG (SEQ ID NO;

14), SDGD (SEQ ID NO;

15), GREDVY (SEQ ID NO;

16), GRGDY (SEQ ID NO;

17), GRGDSP (SEQ ID NO;

18), VAPG (SEQ ID NO;

19), GGGGRGDSP (SEQ ID NO;

20) and GGGGRGDY (SEQ ID NO;

21) and FTLCFD (SEQ ID NO;

22).

30. The system of claim 26 wherein said cells are selected from the group consisting of:

stem cells, muscle cells, pancreatic islets, chondrocytes, hepatic cells, neural cells, renal cortex cells, vascular endothelial cells, thyroid and parathyroid cells, adrenal cells, thymic cells, ovarian cells, and cells which include recombinant genetic material provided to express desired proteins.

31. A composition selected from the group consisting of:

a composition comprising alginate covalently linked to two or more different cell attachment peptides;

a composition comprising alginate mixed with two or more different cell attachment peptides;

a composition comprising alginate covalently linked to one or more cell attachment peptides and mixed with one or more cell attachment peptides;

a composition comprising alginate covalently linked to one cell attachment peptides and ionically linked to chitosan;

a composition comprising alginate covalently linked to two or more different cell attachment peptides and ionically linked to chitosan;

a composition comprising alginate mixed with one cell attachment peptides and ionically linked to chitosan;

a composition comprising alginate mixed with two or more different cell attachment peptides and ionically linked to chitosan

a composition comprising alginate covalently linked to one cell attachment peptide and mixed with cell attachment peptides and ionically linked to chitosan;

a composition comprising alginate covalently linked to two or more different cell attachment peptide and mixed with cell attachment peptides and ionically linked to chitosan;

a composition comprising cells encapsulated in alginate covalently linked to two or more different cell attachment peptides;

a composition comprising cells encapsulated in alginate mixed with two or more different cell attachment peptides;

a composition comprising cells encapsulated in alginate covalently linked to one or more cell attachment peptides and mixed with one or more cell attachment peptides;

a composition comprising cells encapsulated in alginate covalently linked to one cell attachment peptides and ionically linked to chitosan;

a composition comprising cells encapsulated in alginate covalently linked to two or more different cell attachment peptides and ionically linked to chitosan;

a composition comprising cells encapsulated in alginate mixed with one cell attachment peptides and ionically linked to chitosan;

a composition comprising cells encapsulated in alginate mixed with two or more different cell attachment peptides and ionically linked to chitosan

a composition comprising cells encapsulated in alginate covalently linked to one cell attachment peptide and mixed with cell attachment peptides and ionically linked to chitosan; and

a composition comprising cells encapsulated in alginate covalently linked to two or more different cell attachment peptide and mixed with cell attachment peptides and ionically linked to chitosan.

32. The composition of claim 31 wherein the cell attachment peptides are selected from group consisting of:

RGD, YIGSR (SEQ ID NO;

1), IKVAV (SEQ ID NO;

2), REDV (SEQ ID NO;

3), DGEA (SEQ ID NO;

4), VGVAPG (SEQ ID NO;

5), GRGDS (SEQ ID NO;

6), LDV, RGDV (SEQ ID NO;

7), PDSGR (SEQ ID NO;

8), RYVVLPR (SEQ ID NO;

9), LGTIPG (SEQ ID NO;

10), LAG, RGDS (SEQ ID NO;

11), RGDF (SEQ ID NO;

12), HHLGGALQAGDV (SEQ ID NO;

13), VTCG (SEQ ID NO;

14), SDGD (SEQ ID NO;

15), GREDVY (SEQ ID NO;

16), GRGDY (SEQ ID NO;

17), GRGDSP (SEQ ID NO;

18), VAPG (SEQ ID NO;

19), GGGGRGDSP (SEQ ID NO;

20) and GGGGRGDY (SEQ ID NO;

21) and FTLCFD (SEQ ID NO;

22).

33. The composition of claim 31 wherein said composition comprises alginate beads.

34. The composition of claim 31 wherein the beads have mean diameters in a range of from about 30 um to about 500 um.

35. The composition of claim 31 wherein said composition comprises an alginate hydrogel.

36. The composition of claim 31 wherein said composition comprises cells selected from the group consisting of:

stem cells, muscle cells, pancreatic islets, chondrocytes, hepatic cells, neural cells, renal cortex cells, vascular endothelial cells, thyroid and parathyroid cells, adrenal cells, thymic cells, ovarian cells, and cells which include recombinant genetic material provided to express desired proteins.

37. The composition of claim 31 wherein said composition further comprises one or more compounds selected from the group consisting of:

EGF, VEGF, b-FGF, FGF, TGF, TGF-β and

proteoglycans.

38. The composition of claim 31 wherein said composition is in a tissue selected from the group consisting of:

bones, muscle, cartilage, connective, neural, epithelial, vascular, urothelial and mucosal.

39. The composition of claim 31 wherein said composition is in to an organ selected from the group consisting of:

skin, sinus, liver, lungs, kidney, brain, pancreas, endocrine organs, esophagus, stomach, intestines, colon, rectum, cornea, and breast.

40. A method for delivering a desired protein to an individual comprising:

providing to the individual a therapeutically effective amount of a composition selected from the group consisting of one or more of the following;

a composition comprising cells encapsulated in alginate covalently linked to two or more different cell attachment peptides;

a composition comprising cells encapsulated in alginate mixed with two or more different cell attachment peptides;

a composition comprising cells encapsulated in alginate covalently linked to one or more cell attachment peptides and mixed with one or more cell attachment peptides;

a composition comprising cells encapsulated in alginate covalently linked to one cell attachment peptides and ionically linked to chitosan;

a composition comprising cells encapsulated in alginate covalently linked to two or more different cell attachment peptides and ionically linked to chitosan;

a composition comprising cells encapsulated in alginate mixed with one cell attachment peptides and ionically linked to chitosan;

a composition comprising cells encapsulated in alginate mixed with two or more different cell attachment peptides and ionically linked to chitosan

a composition comprising cells encapsulated in alginate covalently linked to one cell attachment peptide and mixed with cell attachment peptides and ionically linked to chitosan; and

a composition comprising cells encapsulated in alginate covalently linked to two or more different cell attachment peptide and mixed with cell attachment peptides and ionically linked to chitosan;

wherein said cells express the desired protein.

41. The method of claim 40 wherein the cell attachment peptides are selected from group consisting of:

RGD, YIGSR (SEQ ID NO;

1), IKVAV (SEQ ID NO;

2), REDV (SEQ ID NO;

3), DGEA (SEQ ID NO;

4), VGVAPG (SEQ ID NO;

5), GRGDS (SEQ ID NO;

6), LDV, RGDV (SEQ ID NO;

7), PDSGR (SEQ ID NO;

8), RYVVLPR (SEQ ID NO;

9), LGTIPG (SEQ ID NO;

10), LAG, RGDS (SEQ ID NO;

11), RGDF (SEQ ID NO;

12), HHLGGALQAGDV (SEQ ID NO;

13), VTCG (SEQ ID NO;

14), SDGD (SEQ ID NO;

15), GREDVY (SEQ ID NO;

16), GRGDY (SEQ ID NO;

17), GRGDSP (SEQ ID NO;

18), VAPG (SEQ ID NO;

19), GGGGRGDSP (SEQ ID NO;

20) and GGGGRGDY (SEQ ID NO;

21) and FTLCFD (SEQ ID NO;

22).

42. The method of claim 40 wherein said composition comprises alginate beads.

43. The method of claim 42 wherein the beads have mean diameters in a range of from about 30 um to about 500 um.

44. The method of claim 40 wherein said composition comprises an alginate hydrogel.

45. The method of claim 40 wherein said composition comprises cells selected from the group consisting of:

stem cells, muscle cells, pancreatic islets, chondrocytes, hepatic cells, neural cells, renal cortex cells, vascular endothelial cells, thyroid and parathyroid cells, adrenal cells, thymic cells, ovarian cells, and cells which include recombinant genetic material provided to express desired proteins.

46. The method of claim 40 wherein said composition further comprises one or more compounds selected from the group consisting of:

EGF, VEGF, b-FGF, FGF, TGF, TGF-β and

proteoglycans.

47. The method of claim 40 wherein said composition is provided as a cross-linked matrix.

48. The method of claim 40 wherein said composition produces a matrix in situ following application to the tissue or organ.

49. The method of claim 40 wherein said individual has been diagnosed with a disease or condition characterized by an absence or deficiency in a functional form of the desired protein.

50. A system for delivering a desired protein to an individual comprising a delivery device for providing a composition to a tissue or organ and a therapeutically effective amount of a composition selected from the group consisting of one or more of the following:

a composition comprising cells encapsulated in alginate covalently linked to two or more different cell attachment peptides;

a composition comprising cells encapsulated in alginate mixed with two or more different cell attachment peptides;

a composition comprising cells encapsulated in alginate covalently linked to one or more cell attachment peptides and mixed with one or more cell attachment peptides;

a composition comprising cells encapsulated in alginate covalently linked to one cell attachment peptides and ionically linked to chitosan;

a composition comprising cells encapsulated in alginate covalently linked to two or more different cell attachment peptides and ionically linked to chitosan;

a composition comprising cells encapsulated in alginate mixed with one cell attachment peptides and ionically linked to chitosan;

a composition comprising cells encapsulated in alginate mixed with two or more different cell attachment peptides and ionically linked to chitosan

a composition comprising cells encapsulated in alginate covalently linked to one cell attachment peptide and mixed with cell attachment peptides and ionically linked to chitosan; and

a composition comprising cells encapsulated in alginate covalently linked to two or more different cell attachment peptide and mixed with cell attachment peptides and ionically linked to chitosan;

wherein said cells express the desired protein.

51. The system of claim 50 wherein said delivery device comprises two lumens.

52. The system of claim 50 wherein said delivery device comprises a syringe.

53. The system of any of claims 50 wherein the cell attachment peptides are selected from group consisting of:

RGD, YIGSR (SEQ ID NO;

1), IKVAV (SEQ ID NO;

2), REDV (SEQ ID NO;

3), DGEA (SEQ ID NO;

4), VGVAPG (SEQ ID NO;

5), GRGDS (SEQ ID NO;

6), LDV, RGDV (SEQ ID NO;

7), PDSGR (SEQ ID NO;

8), RYVVLPR (SEQ ID NO;

9), LGTIPG (SEQ ID NO;

10), LAG, RGDS (SEQ ID NO;

11), RGDF (SEQ ID NO;

12), HHLGGALQAGDV (SEQ ID NO;

13), VTCG (SEQ ID NO;

14), SDGD (SEQ ID NO;

15), GREDVY (SEQ ID NO;

16), GRGDY (SEQ ID NO;

17), GRGDSP (SEQ ID NO;

18), VAPG (SEQ ID NO;

19), GGGGRGDSP (SEQ ID NO;

20) and GGGGRGDY (SEQ ID NO;

21) and FTLCFD (SEQ ID NO;

22).

54. The system of claim 50 wherein said composition comprises alginate beads.

55. The system of claim 54 wherein the beads have mean diameters in a range of from about 30 um to about 500 um.

56. The system of claim 50 wherein said composition comprises an alginate hydrogel.

57. The system of claim 50 wherein said cells are selected from the group consisting of:

stem cells, muscle cells, pancreatic islets, chondrocytes, hepatic cells, neural cells, renal cortex cells, vascular endothelial cells, thyroid and parathyroid cells, adrenal cells, thymic cells, ovarian cells, and cells which include recombinant genetic material provided to express desired proteins.

58. The system of claim 50 wherein said composition further comprises one or more compounds selected from the group consisting of:

EGF, VEGF, b-FGF, FGF, TGF, TGF-β and

proteoglycans.

59. A method of culturing cells in vitro comprising maintaining cells under conditions suitable for cell growth and proliferation in a composition selected from the group consisting of:

a composition comprising alginate covalently linked to two or more different cell attachment peptides;

a composition comprising alginate mixed with two or more different cell attachment peptides;

a composition comprising alginate covalently linked to one or more cell attachment peptides and mixed with one or more cell attachment peptides;

a composition comprising alginate covalently linked to one cell attachment peptides and ionically linked to chitosan;

a composition comprising alginate covalently linked to two or more different cell attachment peptides and ionically linked to chitosan;

a composition comprising alginate mixed with one cell attachment peptides and ionically linked to chitosan;

a composition comprising alginate mixed with two or more different cell attachment peptides and ionically linked to chitosan

a composition comprising alginate covalently linked to one cell attachment peptide and mixed with cell attachment peptides and ionically linked to chitosan; and

a composition comprising alginate covalently linked to two or more different cell attachment peptide and mixed with cell attachment peptides and ionically linked to chitosan.

60. The method of claim 59 wherein the cell attachment peptides are selected from group consisting of:

RGD, YIGSR (SEQ ID NO;

1), IKVAV (SEQ ID NO;

2), REDV (SEQ ID NO;

3), DGEA (SEQ ID NO;

4), VGVAPG (SEQ ID NO;

5), GRGDS (SEQ ID NO;

6), LDV, RGDV (SEQ ID NO;

7), PDSGR (SEQ ID NO;

8), RYVVLPR (SEQ ID NO;

9), LGTIPG (SEQ ID NO;

10), LAG, RGDS (SEQ ID NO;

11), RGDF (SEQ ID NO;

12), HHLGGALQAGDV (SEQ ID NO;

13), VTCG (SEQ ID NO;

14), SDGD (SEQ ID NO;

15), GREDVY (SEQ ID NO;

16), GRGDY (SEQ ID NO;

17), GRGDSP (SEQ ID NO;

18), VAPG (SEQ ID NO;

19), GGGGRGDSP (SEQ ID NO;

20) and GGGGRGDY (SEQ ID NO;

21) and FTLCFD (SEQ ID NO;

22).

61. The method of claim 59 wherein said composition comprises alginate beads.

62. The method of claim 61 wherein the beads have mean diameters in a range of from about 30 um to about 500 um.

63. The method of claim 59 wherein said composition comprises an alginate hydrogel.

64. The method of claim 59 wherein said composition comprises cells selected from the group consisting of:

stem cells, muscle cells, pancreatic islets, chondrocytes, hepatic cells, neural cells, renal cortex cells, vascular endothelial cells, thyroid and parathyroid cells, adrenal cells, thymic cells, ovarian cells, and cells which include recombinant genetic material provided to express desired proteins.

65. The method of claim 59 wherein said composition further comprises one or more compounds selected from the group consisting of:

EGF, VEGF, b-FGF, FGF, TGF, TGF-β and

proteoglycans.