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Last Updated: October 16, 2019

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Claims for Patent: 6,051,425

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Summary for Patent: 6,051,425
Title: Method for producing tissue formation and tissue culture kit
Abstract:A matrix for tissue culture comprising two kinds of sponges having at least one different physical property and/or at least one different chemical property; a method for culturing tissue using said matrix for tissue culture comprising inoculating and culturing first cell on a first sponge, laminating a second sponge thereon, and inoculating and culturing second cell on said second sponge; a method for fixing a cultured tissue comprising placing a cultured tissue in gelatin solution solated by elevating temperature, lowering temperature to gelatinize gelatin to fix the cultured tissue by said gelatinated gelatin; and an artificial skin fixed comprising dermis layer fixed by gelatin and epidermis layer laminated on the dermis layer.
Inventor(s): Morota; Katsuyasu (Ayabe, JP), Morita; Shinichiro (Ayabe, JP)
Assignee: Gunze Limited (Kyoto-fu, JP)
Application Number:08/858,905
Patent Claims:1. A method for producing a tissue formation having a structure differentiated in relation to depth, comprising the steps of:

inoculating a first culturing cell on a first bio-compatible sponge having first pores, said first pores having a size such that said first culturing cell is accommodated in said first pores to allow said first culturing cell to proliferate inside said first bio-compatible sponge;

inoculating a second culturing cell on a second bio-compatible sponge having second pores, said second pores having a size such that said second culturing cell is accommodated on said second bio-compatible sponge where inoculated to allow said second culturing cell to proliferate on said second bio-compatible sponge, said second bio-compatible sponge having a lower degree of resistance against collagenase secreted by said second culturing cell than that of said first bio-compatible sponge against said first culturing cell; and

culturing said first and second culturing cells using at least one culture medium to produce a tissue formation having a structure differentiated in relation to depth.

2. A method according to claim 1, wherein said first bio-compatible sponge is chemically crosslinked to a degree sufficient to prevent decomposition during culture of the cell, and said second bio-compatible sponge is crosslinked by heat-dehydration to a degree sufficient for said second bio-compatible sponge to decompose and dissipate at the conclusion of culture of the cell.

3. A method according to claim 1, wherein said first bio-compatible sponge and said bio-compatible second sponge are each made of a bio-compatible polymer selected from the group consisting of type-I, -II, -III, and -IV collagens, gelatine, sodium alginate, fibronectin, laminin, hyaluronic acid, chitin, chitosan, and EHS mouse tumor solubilized extract.

4. A method according to claim 1, wherein said bio-compatible polymer is type-I, II, -III, or -IV collagen.

5. A method according to claim 1, wherein the pore size of said first pores is 50 .mu.m or more, and the pore size of said second pores is 1-30 .mu.m.

6. A method according to claim 1, wherein said first culturing cell is a human fibroblast cell, and said second culturing cell is a human keratinocyte cell.

7. A tissue culture kit for producing a tissue formation having a structure differentiated in relation to depth, comprising:

a first culturing cell;

a second culturing cell;

a first bio-compatible sponge having first pores, on which said first culturing cell is inoculated, said first pores having a size such that said first culturing cell is accommodated in said first pores to allow said first culturing cell to proliferate inside said first bio-compatible sponge;

a second bio-compatible sponge having second pores, on which said second culturing cell is inoculated, said second pores having a size such that said second culturing cell is accommodated on said second bio-compatible sponge where inoculated to allow said second culturing cell to proliferate on said second bio-compatible sponge, said second bio-compatible sponge having a lower degree of resistance against collagenase secreted by said second culturing cell than that of said first bio-compatible sponge against said first culturing cell; and

at least one culture medium for culturing said first cell and second cell.

8. A tissue culture kit according to claim 7, wherein said first bio-compatible sponge is chemically crosslinked to a degree sufficient to prevent decomposition during culture of the cell, and said second bio-compatible sponge is crosslinked by heat-dehydration to a degree sufficient for said second bio-compatible sponge to decompose and dissipate at the conclusion of culture of the cell.

9. A tissue culture kit according to claim 7, wherein said first bio-compatible sponge and said bio-compatible second sponge are each made of a bio-compatible polymer selected from the group consisting of type-I, -II, -III, and -IV collagens, gelatine, sodium alginate, fibronectin, laminin, hyaluronic acid, chitin, chitosan, and EHS mouse tumor solubilized extract.

10. A tissue culture kit according to claim 9, wherein said bio-compatible polymer is type-I, -II, -III, or -IV collagen.

11. A tissue culture kit according to claim 9, wherein the pore size of said first pores is 50 .mu.m or more, and the pore size of said second pores is 1-30 .mu.m.

12. A tissue culture kit according to claim 9, wherein said first culturing cell is a human fibroblast cell, and said second culturing cell is a human keratinocyte cell.

13. A tissue culture kit for a tissue formation having a structure differentiated in relation to depth, comprising a bio-compatible matrix essentially consisting of:

a first bio-compatible sponge adapted to be inoculated with a first culturing cell, said first bio-compatible sponge having a thickness of 1-5 mm and having first pores having a size of at least 50 .mu.m such that the first culturing cell is accommodated in the first pores, said first bio-compatible sponge being chemically crosslinked to a degree sufficient to prevent decomposition during culture of the cell; and

a second bio-compatible sponge adapted to be inoculated with a second culturing cell, said second bio-compatible sponge having a thickness of 1-2 mm and having second pores having a size of 1-30 .mu.m such that the second culturing cell is accommodated on said second bio-compatible sponge where inoculated, said second bio-compatible sponge being crosslinked by heat-dehydration to a degree sufficient for said second bio-compatible sponge to decompose and dissipate at the conclusion of culture of the cell.

14. A tissue culture kit according to claim 13, wherein said first bio-compatible sponge and said second bio-compatible sponge are made of collagen.

15. A tissue culture kit according to claim 13, wherein said bio-compatible sponge matrix consists of said first bio-compatible sponge and said second bio-compatible sponge.

Summary for Patent:   Start Trial

Foriegn Application Priority Data
Foreign Country Foreign Patent Number Foreign Patent Date
Japan6-263047Sep 19, 1994

Details for Patent 6,051,425

Applicant Tradename Biologic Ingredient Dosage Form BLA Number Approval Date Patent No. Assignee Estimated Patent Expiration Status Orphan Source
Advance Biofactures SANTYL collagenase VIAL 101995 001 1965-06-04   Start Trial Gunze Limited (Kyoto-fu, JP) 2014-09-19 RX search
>Applicant >Tradename >Biologic Ingredient >Dosage Form >BLA >Number >Approval Date >Patent No. >Assignee >Estimated Patent Expiration >Status >Orphan >Source

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