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The present invention relates to cell adhesion promoting (“CAP”) peptide combinations that promote cell attachment or cell adhesion to culture surfaces that are otherwise cell adhesion resistant “CAR”. The invention provides combination of peptides that, when covalently coupled to a CAR layer such as hyaluronic acid that has been created on a polystyrene surface, promote cell attachment, growth differentiation, and execution of other desired cellular functions in culture.

Inventors: Richard David Guarino, Bryce Nelson Chaney, Andrea Liebmann-Vinson, Mohammad A. Heidaran
Original Assignee: Becton, Dickinson and Company
Primary Examiner: Maryam Monshipouri
Secondary Examiner: Marsha M Tsay
Attorney: Alston & Bird LLP
Current U.S. Classification: 435/289.1; 435/402; 435/404; 530/330; 530/402

View patent at USPTO
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Citations

Cited PatentFiling dateIssue dateOriginal AssigneeTitle
US4851521Jul 2, 1986Jul 25, 1989Fidia, S.p.A.Esters of hyaluronic acid
US5278063Sep 28, 1989Jan 11, 1994Board of Regents The University of Texas SystemChemical modification of promote animal cell adhesion on surfaces
US6150459Apr 13, 1999Nov 21, 2000Massachusetts Institute of TechnologyComb polymers for regulating cell surface interactions
US20030022152Mar 5, 2002Methods, apparatus and computer program products for formulating culture media
US20030113812Sep 30, 2002Proliferation and differentiation of stem cells using extracellular matrix and other molecules
US20030113813Sep 30, 2002Methods and devices for the integrated discovery of cell culture environments
US20030162289Nov 19, 2001Peptides promoting cell adherence, growth and secretion
US20030175745Sep 30, 2002Peptides with growth inhibitory action

Claims

1. A cell culture vessel for culturing adherent cells, said vessel comprising a polymer surface, a hyaluronic acid (HA) layer bonded to said polymer surface, and one or more peptides covalently bound to the HA layer, wherein said peptides promote adherence of cells that do not adhere to the HA layer in the absence of said peptides, and wherein each of said one or more peptides comprises:

(a) one or more peptide sequences selected from the group consisting of SEQ ID NO:2 (EFIKY), SEQ ID NO:3 (EEEKV), SEQ ID NO:4 (GHK), SEQ ID NO:6 (KKLVY), SEQ ID NO:8 (AFAFK), SEQ ID NO:10 (KMLIY), SEQ ID NO:11 (KSYYY), SEQ ID NO:12 (AKKMV), SEQ ID NO:14 (AFIPV), SEQ ID NO:15 (KKMYY), and SEQ ID NO:16 (AIKKK);

(b) a chemically or recombinantly prepared peptide multimer that includes at least two repeats of any one or a combination of SEQ ID NOs:2-4, 6, 8, 10-12, and 14-16; or

(c) a cell adhesion promoting variant of any one or more of SEQ ID NOs:2-4, 6, 8, 10-12, and 14-16, said variant having a conservative substitution of a single amino acid residue, wherein said variant has the ability to promote cell attachment or growth.

2. The cell culture vessel of claim 1, wherein said cells are fibroblasts or osteoblasts.

3. The cell culture vessel of claim 1, wherein said cells are liver cells.

4. The cell culture vessel of claim 1, wherein said cells are human or rat primary hepatocytes or porcine hepatocytes.

5. The cell culture vessel of claim 1, wherein said cell culture vessel is a multiwell plate.

6. The cell culture vessel of claim 1, wherein said polymer surface is formed from a flexible material.

7. The system cell culture vessel of claim 1, wherein said polymer surface is selected from the group consisting of polystyrene, polypropylene, polyethylene tereplithalate, polylactide, cellulose, and combinations thereof.

8. The cell culture vessel of claim 7, wherein said polymer surface is polystyrene.

9. A cell culture vessel for culturing adherent liver cells, said vessel comprising a polymer surface, a cell adhesion resistant (CAR) material bonded to said polymer surface, and one or more peptides covalently bound to the CAR material, wherein said peptides promote adherence of liver cells that do not adhere to the CAR material in the absence of said peptides, and wherein each of said one or more peptides comprises:

(a) one or more peptide sequences selected from the group consisting of SEQ ID NO:2 (EFIKY), SEQ ID NO:3 (EEEKV), SEQ ID NO:4 (GHK), SEQ ID NO:6 (KKLVY), SEQ ID NO:8 (AFAFK), SEQ ID NO:10 (KMLIY), SEQ ID NO: 11 (KSYYY), SEQ ID NO:12 (AKKMV), SEQ ID NO:14 (AFIPV), SEQ ID NO:15 (KKMYY), and SEQ ID NO:16 (AIKKK);

(b) a chemically or recombinantly prepared peptide multimer that includes at least two repeats of any one or a combination of SEQ ID NOs:2-4, 6, 8, 10-12, and 14-16; or

(c) a cell adhesion promoting variant of any one or more of SEQ ID NOs:2-4, 6, 8, 10-12, and 14-16, said variant having a conservative substitution of a single amino acid residue, wherein said variant has the ability to promote liver cell attachment or growth.

10. The cell culture vessel of claim 9, wherein said cell adhesion resistant (CAR) material is selected from the group consisting of hyaluronic acid (HA), alginic acid (AA), polyhydroxyethyl methacrylate (Poly-HEMA), polyethylene glycol (PEG), glyme, polyisopropylacrylamide, and combinations thereof.

11. The cell culture vessel of claim 10, wherein said cell adhesion resistant (CAR) material is hyaluronic acid (HA).

12. The cell culture vessel of claim 9 wherein said liver cells are human or rat primary hepatocytes or porcine hepatocytes.

13. The cell culture vessel of claim 9, wherein said cell culture vessel is a multiwell plate.

14. The cell culture vessel of claim 9, wherein said polymer surface is formed from a flexible material.

15. The cell culture vessel of claim 9, wherein said polymer surface is selected from the group consisting of polystyrene, polypropylene, polyethylene terephthalate, polylactide, cellulose, and combinations thereof.

16. The cell culture vessel of claim 15, wherein said polymer surface is polystyrene.

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