Transplantable sheets of living keratinous tissue

• US 4,304,866 A
• Filed: 11/14/1979
• Issued: 12/08/1981
• Est. Priority Date: 11/14/1979
• Status: Expired due to Term

• Current Assignee: Massachusetts Institute of Technology
• Original Assignee: Massachusetts Institute of Technology
• Inventors: Green, Howard, Kehinde, Olaniyi
• Primary Examiner(s): Kepplinger, Esther M.
• Application Number:
  US06/094,003
• Time in Patent Office: 755 Days
• Field of Search: 435/240, 435/241, 435/267, 424/95, 128/1, 128/305.5
• US Class Current: 424/574
• CPC Class Codes: A61K 35/12 Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells vaccines or medicinal preparations containing antigens or antibodies A61K39/00
• CPC Class Codes: A61K 35/36 Skin; Hair; Nails; Sebaceous glands; Cerumen; Epidermis; Epithelial cells; Keratinocytes; Langerhans cells; Ectodermal cells islets of Langerhans A61K35/39
• CPC Class Codes: A61L 27/60 Materials for use in artificial skin
• CPC Class Codes: C12N 2509/00 Methods for the dissociation of cells, e.g. specific use of enzymes
• CPC Class Codes: C12N 5/0629 Keratinocytes; Whole skin
• CPC Class Codes: Y02A 50/30 Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
• CPC Class Codes: Y10S 623/915 Method or apparatus for preparing biological material

Abstract

A method of producing transplantable sheets of living keratinous tissue by culturing keratinocytes in a culture vessel and subsequently enzymatically detaching a sheet of keratinous tissue employing an enzyme, such as Dispase, is disclosed herein.

Claims

1. A method of producing transplantable sheets of living keratinous tissue, comprising:

a. culturing keratinocytes in a culture vessel under conditions whereby a sheet of keratinous tissue is formed upon the surface of the vessel, and

b. incubating said sheet of keratinous tissue in the presence of a neutral protease under conditions sufficient to enzymatically detach the sheet of keratinous tissue from the surface of the vessel without disaggregating said sheet.

2. A method of claim 1 wherein the keratinocytes comprise human epidermal cells.

3. A method of claim 2 wherein the culturing of keratinocytes is performed in the presence of fibroblast cells treated to prevent their multiplication or medium sufficiently conditioned by fibroblast cells to allow keratinocyte colonies to grow.

4. A method for treating defects in the epithelium of a mammal, comprising:

a. obtaining living epidermal cells from said mammal;

b. culturing said epidermal cells in a culture vessel to form a sheet of epidermal tissue in said vessel;

c. incubating said sheet of epidermal tissue in the presence of a neutral protease under conditions sufficient to enzymatically detach said tissue from the culture vessel without disaggregating the epidermal cells; and

,

d. transplanting detached epidermal tissue onto the defect of said mammal.

5. A method of claim 4 wherein the mammal comprises a human being.