Skin culture and process for preparing the same

• US 6,043,089 A
• Filed: 05/11/1998
• Issued: 03/28/2000
• Est. Priority Date: 03/07/1995
• Status: Expired due to Fees

• Current Assignee: Menicon Company Limited
• Original Assignee: Menicon Company Limited
• Inventors: Moriyama, Takeshi, Hamano, Kyoko, Sugiyama, Akihisa
• Primary Examiner(s): Wortman, Donna C.
• Assistant Examiner(s): Brumback, Brenda G.
• Application Number:
  US09/075,519
• Time in Patent Office: 687 Days
• Field of Search: 435/347, 435/373, 435/395, 435/397, 435/371, 435/283.1, 435/284.1, 435/287.1, 435/288.3, 623/15
• US Class Current: 435/371
• CPC Class Codes: C12M 21/08 for producing artificial tissue or for ex-vivo cultivation of tissue prostheses A61F2/00, grafts A61L27/00
• CPC Class Codes: C12M 25/14 Scaffolds; Matrices in general C12N5/0068
• CPC Class Codes: C12N 2502/094 keratinocytes
• CPC Class Codes: C12N 2502/1323 Adult fibroblasts
• CPC Class Codes: C12N 2533/54 Collagen; Gelatin
• CPC Class Codes: C12N 5/0068 General culture methods using substrates for specific animal cell type C12N5/06
• CPC Class Codes: C12N 5/0698 Skin equivalents

Abstract

A process for preparing a skin culture which comprises the following steps:

a) a step in which a skin culture matrix comprising a collagen sponge, a collagen sheet or a collagen gel is prepared in a means having a projection to provide said matrix with a penetrating pore,

b) a step in which a skin-derived cell is seeded and cultured on said matrix, and

c) a step in which the skin culture is provided with a penetrating pore by the means having the projection before said skin culture covers a skin defect, if necessary, the steps a) and b) are simultaneously carried out; a skin culture matrix and a skin culture obtained thereby. According to the present invention, a skin culture can be prepared wherein sufficient nutrition supply to epidermal cells and/or fibroblasts cultured and proliferated and discharge of exudate excessively pooled in the wound can be smoothly performed, while contraction is inhibited, by producing a penetrating pore in the culture skin before covering the wound with the culture skin with a means which requires no pretreatment in order to prevent the flowing out and losing of cells which are caused in seeding cells on a collagen matrix.

Claims

1. A skin cell culture which comprises:

(1) a skin cell culture matrix comprising a collagen sponge, a collagen sheet or a collagen gel, said matrix being prepared in a structure provided with a plurality of projections, wherein said matrix has a thickness of at least 1 mm, and each of said projections has a width of at least 1 mm, said width being measured in at least one direction across a cross-section of said projections, and

(2) skin-derived cells seeded and cultured on said matrix in said structure provided with a plurality of projections,

wherein the resulting matrix bearing a culture of skin-derived cells is provided with a plurality of penetrating pores by the operation of said structure provided with a plurality of projections immediately before said skin cell culture is used to cover a skin defect.

2. The skin cell culture of claim 1, wherein said structure provided with a plurality of projections is a support plate provided with said projections, or a vessel with a bottom of which the inner surface is provided with said projections.

3. The skin cell culture of claim 2, wherein said support plate or said bottom of said vessel is fitted with a removable sheet having perforations matching said projections, and said skin cell culture is provided on said removable sheet.

4. The skin cell culture of claim 1, wherein skin-derived fibroblasts are seeded and cultured on one side or both sides of said matrix.

5. The skin cell culture of claim 1, wherein epidermal cells are seeded and cultured on one side of said matrix.

6. The skin cell culture of claim 1, wherein skin-derived fibroblasts are seeded and cultured on one side of both sides of said matrix and epidermal cells are seeded and cultured on one side of said matrix.

7. The skin cell culture of any one of claims 1-3, wherein the collagen is atelocollagen.

8. The skin cell culture of any one of claims 1-3, wherein the skin cell culture matrix contains hyaluronic acid.