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US5015584A Expired / Lapsed US ⏱️ 0.0 years remaining

Epidermal graft system

Assignee: The University of Texas System
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Filing: 1988-02-09
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Granted: 1991-05-14
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Expires: 2008-02-09
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Abstract

Abstract The present invention involves a readily produced skin autograft or allograft composite suitable for the supplementation or replacement of injured skin. A sheet of collagen-coated pliable material such as synthetic surgical dressing is used as a foundation for the skin autograft composite. The synthetic surgical dressing, if not precoated with collagen, may be placed in a container and coated with collagen. Epidermal cells, preferably obtained from a prospective recipient of the skin autograft composite being produced, are cultured in an appropriate medium substantially preventing cell differentiation to form epidermal cells on the surface of a collagen-coated container. When substantially confluent, the epidermal cells are enzymatically detached from the culture vessel and layered upon a collagen coated desiccated surgical dressing infused with low calcium culture medium. In actual use, the skin autograft composite is inverted, placed on a recipient, and allowed to remain in position until epidermal cells attach to the recipient and facilitate the formation of a skin-like covering.

Inventors

  • 👤 Miriam M. Brysk

Bibliographic Details

Application Number —
Jurisdiction US
Data Source google_patents
Last Synchronized 2026-10-24 16:31
CPC Classifications
A; A61; A61L; A61L27/00; A61L27/14; A61L27/22; A61L27/24; A61F; A61F2/00; A61F2/02; A61F2/10; A61L27/36; A61L27/50; A61L27/60; A61F2310/00; A61F2310/00005; A61F2310/00365

Claims Analysis

Total Claims: 25
Claim 1
Independent Claim 55 words
A process for the preparation of a skin autograft composite, the process comprising: obtaining human epidermal cells by cultivation in a vessel containing MCDB-153; and attaching the epidermal cells to a collagen coated surgical dressing immmersed in eucaryotic cell cultrure medium comprising MCDB-153 to product a skin autograft composite consisting essentially of unidifferentiated epidermal cells.
Claim 2
Dependent Claim 55 words
A process for the preparation of a skin autograft composite, the process comprising: obtaining human epidermal cells by cultivation in a vessel containing MCDB-153; and attaching the epidermal cells to a collagen coated surgical dressing immmersed in eucaryotic cell cultrure medium comprising MCDB-153 to product a skin autograft composite consisting essentially of unidifferentiated epidermal cells.
Claim 3
Dependent Claim 21 words
The process of claim 1 wherein the eucaryotic cell culture medium promotes epidermal cell attachment to the collagen coated surgical dressing.
Claim 4
Dependent Claim 25 words
The process of claim 1 wherein the human epidermal cells are those of a patient to be treated with the skin autograft composite bieng prepared.
Claim 5
Dependent Claim 22 words
The process of claim 1 wherein the surgical dressing is synthetic and comprises membranous polyurethane, polyethylene oxide/water, hydrocolloid, silicone/nylon or monofilament nylon.
Claim 6
Dependent Claim 16 words
The process of claim 1 wherein the surgical dressing is synthetic and comprises membranous polyethylene oxide/water.
Claim 7
Dependent Claim 122 words
A process for the preparation of a skin autograft composite, the process comprising: obtaining human epidermal cells by cultivation in a vessel containing MCDB-153; adding the epidermal cells to a container comprising a collagen-coating immersed in a cell attachment eucaryotic cell medium and then replacing said cell attachment medium with MCDB-153; maintaining the cells in a an unidifferentiated state in the container during incubation to facilitate growth; subculturing the cells by detachment and incubation after reinoculation in a collagen-coated container with MCDB-153; detaching the subcultured cells and inoculating them in a container with a surface of collagen-coated synthetic surgical dressing immersed in medium comprising MCDB-153; and facilitating undifferentiated cell growth on the collagen-coated synthetic surgical dressing to produce a skin autograft composite.
Claim 8
Dependent Claim 122 words
A process for the preparation of a skin autograft composite, the process comprising: obtaining human epidermal cells by cultivation in a vessel containing MCDB-153; adding the epidermal cells to a container comprising a collagen-coating immersed in a cell attachment eucaryotic cell medium and then replacing said cell attachment medium with MCDB-153; maintaining the cells in a an unidifferentiated state in the container during incubation to facilitate growth; subculturing the cells by detachment and incubation after reinoculation in a collagen-coated container with MCDB-153; detaching the subcultured cells and inoculating them in a container with a surface of collagen-coated synthetic surgical dressing immersed in medium comprising MCDB-153; and facilitating undifferentiated cell growth on the collagen-coated synthetic surgical dressing to produce a skin autograft composite.
Claim 9
Dependent Claim 104 words
A process for the preparation of a skin autograft composite, the process comprising: obtaining human epidermal cells by a process involving treatment of a epidermal split-thickness human skin section with aqueous solution comprising a proteolytic enzyme to separate dermis and epidermis and separating epidermal cells from the section; adding the epidermal cells to a collagen-coating immersed in a container containing a cell attachment eucaryotic cell medium comprising MCDB-153; subculturing the epidermal cells in an undifferentiated state to facilitate cell growth; and growing the subcultured epidermal cells in medium comprising MCDB-153 and on a collagen-coated synthetic surgical dressing to result in a skin autograft composite.
Claim 10
Dependent Claim 104 words
A process for the preparation of a skin autograft composite, the process comprising: obtaining human epidermal cells by a process involving treatment of a epidermal split-thickness human skin section with aqueous solution comprising a proteolytic enzyme to separate dermis and epidermis and separating epidermal cells from the section; adding the epidermal cells to a collagen-coating immersed in a container containing a cell attachment eucaryotic cell medium comprising MCDB-153; subculturing the epidermal cells in an undifferentiated state to facilitate cell growth; and growing the subcultured epidermal cells in medium comprising MCDB-153 and on a collagen-coated synthetic surgical dressing to result in a skin autograft composite.
Claim 11
Dependent Claim 101 words
A process for preparation of a skin autograft composite comprising collagen-coated synthetic surgical dressing and epidermal cells, the process comprising: adding human epidermal cells to a culture vessel containing MCDB-153 facilitating maintenance of undifferentiated cells to produce an inoculum having a cell density of between about 5×10 3 cells per cm 2 and about 10 5 cells per cm 2 and growing the cells to a state of substantial confluence; and incubating the inoculum in a container with a eucaryotic cell attachment medium comprising MCDB-153 to facilitate undifferentiated cell attachment to a sheet of collagen-coated synthetic surgical dressing in said container.
Claim 12
Dependent Claim 101 words
A process for preparation of a skin autograft composite comprising collagen-coated synthetic surgical dressing and epidermal cells, the process comprising: adding human epidermal cells to a culture vessel containing MCDB-153 facilitating maintenance of undifferentiated cells to produce an inoculum having a cell density of between about 5×10 3 cells per cm 2 and about 10 5 cells per cm 2 and growing the cells to a state of substantial confluence; and incubating the inoculum in a container with a eucaryotic cell attachment medium comprising MCDB-153 to facilitate undifferentiated cell attachment to a sheet of collagen-coated synthetic surgical dressing in said container.
Claim 13
Dependent Claim 25 words
The process of claim 8 wherein the eucaryotic cell attachment medium comprises a 1:1 mixture of EMEM with 10% fetal calf serum and MCDB -153.
Claim 14
Dependent Claim 17 words
The process of claim 8, wherein the eucaryotic cell attachment medium further comprises Eagle's Minimal Essential Medium.
Claim 15
Dependent Claim 30 words
The process of claim 8 wherein the eucaryotic cell attachment medium further comprises Eagle's Minimal Essential Medium with 10% fetal calf serum, 20 mM L-glutamine, antibiotics and 0.4 μg/ml hydrocortisone.
Claim 16
Dependent Claim 92 words
A process for the preparing of a skin autograft composite for use on skin injury of a burn victim, the process comprising: obtaining human epidermal cells from said victim and culturing said cells in a vessel comprising MCDB-153 until substantially confluent; transferring the cells to a sheet of collagen-coated synthetic surgical dressing infused with a eucaryotic cell attachment medium comprising MCDB-153 and maintaining the cells in an undifferentiated condition; and removing from the medium the sheet of collagen-coated synthetic surgical dressing having attached undifferentiated cells, said sheet being a skin autograft composite.
Claim 17
Dependent Claim 92 words
A process for the preparing of a skin autograft composite for use on skin injury of a burn victim, the process comprising: obtaining human epidermal cells from said victim and culturing said cells in a vessel comprising MCDB-153 until substantially confluent; transferring the cells to a sheet of collagen-coated synthetic surgical dressing infused with a eucaryotic cell attachment medium comprising MCDB-153 and maintaining the cells in an undifferentiated condition; and removing from the medium the sheet of collagen-coated synthetic surgical dressing having attached undifferentiated cells, said sheet being a skin autograft composite.
Claim 18
Dependent Claim 76 words
A process for the preparing of a skin autograft or allograft composite, the process comprising: placing a sheet of collagen-coated synthetic surgical dressing in a container; obtaining human epidermal cells; adding a quantity of eucaryotic cell attachment medium which comprises MCDB-153 and the human epidermal cells to the container; and culturing the epidermal cells to facilitate attachment to the collagen-coated synthetic surgical dressing and proliferation of undifferentiated cells to produce a skin autograft or allograft composite.
Claim 19
Dependent Claim 76 words
A process for the preparing of a skin autograft or allograft composite, the process comprising: placing a sheet of collagen-coated synthetic surgical dressing in a container; obtaining human epidermal cells; adding a quantity of eucaryotic cell attachment medium which comprises MCDB-153 and the human epidermal cells to the container; and culturing the epidermal cells to facilitate attachment to the collagen-coated synthetic surgical dressing and proliferation of undifferentiated cells to produce a skin autograft or allograft composite.
Claim 20
Dependent Claim 53 words
The process of claim 13 wherein the placing step includes the steps of: placing a sheet of synthetic surgical dressing on the lower surface of a container; treating the placed synthetic surgical dressing with an aqueous solution of collagen and evaporating water from said solution to produce a modified collagen-coated synthetic surgical dressing.
Claim 21
Dependent Claim 27 words
The process of claim 13 wherein the sheet of collagen-coated synthetic surgical dressing is defined further as comprising membranous polyurethane, polyethylene oxide/water, hydrocolloid, silicone/nylon or monofilament nylon.
Claim 22
Dependent Claim 20 words
The process of claim 13 wherein the sheet of collagen-coated synthetic surgical dressing is defined as comprising membranous polyethylene oxide/water.
Claim 23
Dependent Claim 66 words
The process of claim 13 wherein obtaining human epidermal cells id defined further as including the steps of: obtaining a human skin section; washing the human skin section with eucaryotic cell attachment medium comprising MCDB-153; rinsing the human skin section with isotonic buffered saline free of divalent cations; treating the human skin section with an aqueous solution comprising a proteolytic enzyme; and separating dissociated epidermal cells.
Claim 24
Dependent Claim 73 words
A process for the production of a skin autograft composite, the process comprising; obtaining epidermal cells from a patient; allowing said cells to proliferate in a culture vessel by in vitro attachment and maintenance in an undifferentiated state until substantially confluent; and transferring the epidermal cells to a collagen-located sheet of surgical dressing emplaced in a container and immersed in differentiation-impeding eucaryotic cell attachment medium comprising MCDB-153 to produce a skin autograft composite.
Claim 25
Dependent Claim 73 words
A process for the production of a skin autograft composite, the process comprising; obtaining epidermal cells from a patient; allowing said cells to proliferate in a culture vessel by in vitro attachment and maintenance in an undifferentiated state until substantially confluent; and transferring the epidermal cells to a collagen-located sheet of surgical dressing emplaced in a container and immersed in differentiation-impeding eucaryotic cell attachment medium comprising MCDB-153 to produce a skin autograft composite.

References Cited (Backward Citations)

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USPTO Annuity & Maintenance Windows

Annuity Stage Window Opens Payment Deadline Surcharge Period Until Estimated Fee Status
3.5 Years (Year 4) 1994-05-13 1994-11-12 1995-05-14 $2000.00 Window Closed / Lapsed
7.5 Years (Year 8) 1998-05-13 1998-11-12 1999-05-14 $3760.00 Window Closed / Lapsed
11.5 Years (Year 12) 2002-05-13 2002-11-12 2003-05-14 $7700.00 Window Closed / Lapsed

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