Cultured Melanocytes on Bioploymer Membranes for Treatment of Hyper and Hypopigmentation Disorders

• US 20080089870A1
• Filed: 10/15/2007
• Published: 04/17/2008
• Est. Priority Date: 10/13/2006
• Status: Abandoned Application

• Current Assignee: Reliance Life Sciences Pvt. Ltd.
• Original Assignee: Reliance Life Sciences Pvt. Ltd.
• Inventors: Shenoy, Sudheer, Ghosh, Deepa, Shah, Viru, Kuchroo, Pushpa
• Application Number:
  US11/872,431
• Publication Number:
  US 20080089870A1
• Time in Patent Office: Days
• US Class Current: 424/93.7
• CPC Class Codes: A61L 27/3813 Epithelial cells, e.g. keratinocytes, urothelial cells
• CPC Class Codes: A61L 27/60 Materials for use in artificial skin
• CPC Class Codes: A61P 17/00 Drugs for dermatological disorders
• CPC Class Codes: C12N 2502/091 melanocytes
• CPC Class Codes: C12N 2502/094 keratinocytes
• CPC Class Codes: C12N 2502/1323 Adult fibroblasts
• 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 5/0626 Melanocytes
• CPC Class Codes: C12N 5/0698 Skin equivalents

Abstract

The present invention relates to a graft wherein cultured autologous melanocytes are delivered using a biopolymer. The present invention describes the composition, method of preparation and its properties relating to safety and efficacy. The graft of the present invention has a potential use in treating hyper- and hypopigmentation disorders.

Claims

1. A graft composition capable of inducing pigmentation in skin comprising melanocyte cells derived from autologous epidermis and a biopolymer membrane, wherein at least 75% of the cells remain viable in the composition for at least 72 hours under transport conditions at 5-37°

C.

2. The composition of claim 1, wherein, wherein at least 80% of the cells remain viable in the composition for at least 96 hours under transport conditions at 5-37°

C.

3. The composition of claim 1, wherein the composition further comprises keratinocyte cells.

4. The composition of claim 3, wherein the composition the melanocyte and keratinocyte cells are present in a ratio between 0.8:

2 to 1;

1 on the biopolymer membrane.

5. The composition of claim 1, wherein the cells are present as a monolayer on the biopolymer membrane.

6. The composition of claim 5, wherein said monolayer is a sub-confluent monolayer.

7. The composition of claim 1, wherein the melanocytes cells are capable of further proliferation.

8. The composition of claim 1, wherein at least 90% of the cells are actively proliferating melanocytes.

9. The composition of claim 1, wherein the biopolymer membrane selected from the group consisting of polylactic acid (PLA), polyglycolic acid (PGA), and polylactic-polyglycolic acid copolymer (PLGA) having a molecular weight of at least 50,000 Da.

10. The composition of claim 1, wherein the biopolymer membrane is polylactic acid (PLA).

11. The composition of claim 1, wherein viable melanocyte cells are present in a sufficient quantity so that the resulting composition has a therapeutic effect against a hypo-pigmentation or hyper-pigmentation disorder of the skin.

12. A process for preparing the composition of claim 1 comprising the steps of:

a) isolating melanocyte cells from autologous epidermis;

b) preparing a cell suspension comprising the isolated melanocyte cells;

c) expanding the melanocyte cells;

d) optionally, cryopreserving the melanocyte cells;

e) optionally, thawing the melanocyte cells;

f) seeding the melanocyte cells onto a biopolymer membrane;

g) expanding the cells into a sub-confluent monolayer on the biopolymer membrane; and

h) transporting the biopolymer membrane using a transport device comprising transport media;

wherein at least 75% of the cells remain viable in the composition for at least 72 hours under transport conditions.

13. The process of claim 12, wherein the biopolymer membrane comprises polylactic acid (PLA), and the melanocyte cells are seeded onto the biopolymer membrane at a cell concentration of 1×

10⁴ to 1×

10⁵ cells/cm².

14. The process of claim 12, wherein the transport device comprises polycarbonate.

15. The process of claim 12, wherein the transport media comprises carbon dioxide enriched media.

16. The process of claim 12, wherein the transport media is KSFM media.

17. A method for treating an individual requiring skin pigmentation comprising applying the composition of claim 1 to the individual, wherein the composition enhances the rate of epidermal regeneration in the skin of the individual.

18. The method of claim 17, wherein the individual is human.

19. The method of claim 17, wherein the individual requiring skin pigmentation is suffering from vitiligo.

20. A composition comprising a sub-confluent monolayer of melanocyte cells and a biopolymer membrane, wherein at least 75% of the cells remain viable in the composition for at least 72 hours under transport conditions at 5-37°

C.

21. A composition comprising a subconfluent monolayer of melanocytes co-cultured with keratinocytes and a biopolymer membrane, wherein at least 75% of the melanocyte cells remain viable in the composition for at least 72 hours under transport conditions at 5-37°

C.

22. A composition of claim 21 wherein the melanocytes is cocultured in a ratio of 1:

1 with keratinocytes.

23. A composition comprising (1) a sub-confluent monolayer of keratinocyte cells comprising undifferentiated cells and (2) a biopolymer membrane, wherein at least 75% of the cells remain viable in the composition for at least 72 hours under transport conditions at 5-37°

C.

24. A method for treating hypopigmentation in a subject, comprising using a biopolymer membrane as a delivery system, and delivering to the skin a sub-confluent monolayer of melanocytes.

25. A method for treating hyperpigmentation in a subject, comprising using a biopolymer membrane as a delivery system, and delivering to the skin a sub-confluent monolayer of melanocytes

26. A kit for treating vitiligo, wherein the kit comprises (1) a composition comprising (a) a sub-confluent monolayer of melanocyte cells and (b) a biopolymer membrane, wherein the monolayer is on the biopolymer membrane;

and (2) a transport device,

wherein at least 75% of the melanocyte cells remain viable in the composition for at least 72 hours under transport conditions at 5-37°

C.