System and method for fabricating a cornea
Inventors
Moss, Sarah • Hoying, James • Strobel, Hannah
Assignees
Interested in licensing this patent?
MTEC can help explore whether this patent might be available for licensing for your application.
Publication Number
US-11571496-B2
Publication Date
2023-02-07
Expiration Date
Abstract
A method for fabricating a cornea includes affixing a frame to at least one cell culture insert comprising a generally cylindrical structure having a proximal end and a distal end, a base disposed at the proximal end, and a porous membrane disposed between the proximal end and the distal end; affixing a dome-shaped member to the porous membrane within the frame, the dome-shaped member comprising a crown, a dome base, and a surface connecting the crown and the dome base; depositing a material comprising a matrix-forming compound on the frame such that the crown and at least a portion of the surface of the dome-shaped member is coated with the material comprising the matrix-forming compound; and removing the dome-shaped member to produce a fabricated cornea attached to the frame. A system for fabricating a cornea and a cornea scaffold are also described herein.
Core Innovation
The invention describes a system for fabricating a cornea using at least one generally cylindrical cell culture insert having a proximal end, a distal end, a base disposed at the proximal end, and a porous membrane disposed between the proximal end and the distal end. A frame is disposed on the at least one cell culture insert, and a generally dome-shaped member is disposed on the porous membrane, where the dome-shaped member comprises a crown, a dome base, and a surface connecting the crown and the dome base.
The dome-shaped member is positioned such that the surface projects from the porous membrane, with the crown extending away from the base disposed at the proximal end and beyond the distal end of the cell culture insert. The dome-shaped member comprises a sacrificial material, and a fabricated cornea is produced in which a matrix-forming compound is deposited on the frame and the dome-shaped member so that the crown and at least a portion of the surface of the dome-shaped member are coated with the matrix-forming compound.
The documented system and method further describe use of a sacrificial dome-shaped member that is coated with a matrix-forming compound, after which the sacrificial material is removed to yield a fabricated cornea/scaffold attached to the frame. The cellular and matrix composition described includes corneal keratocytes, corneal endothelium cells, and corneal epithelium cells, in connection with the matrix-forming compound deposited on the dome-shaped member.
Claims Coverage
The independent claim specifies four inventive features centered on a cornea-fabricating system with a cylindrical cell culture insert including a porous membrane, a frame, and a sacrificial dome-shaped member that projects beyond the insert, followed by matrix-forming-compound deposition to coat the crown and dome surface. Dependent claims further refine the sacrificial material, define an insert-holding assembly, specify fluid inlet/outlet lines, and enumerate candidate matrix-forming compounds.
Cylindrical cell culture insert with porous membrane
At least one cell culture insert having a generally cylindrical structure with a proximal end and a distal end, a base disposed at the proximal end, and a porous membrane disposed between the proximal end and the distal end.
Frame disposed on the cell culture insert
A frame disposed on the at least one cell culture insert.
Sacrificial dome-shaped member protruding beyond the insert
A generally dome-shaped member disposed on the porous membrane, comprising a crown, a dome base, and a surface connecting the crown and the dome base, with the surface projecting from the porous membrane such that the crown extends away from the base and beyond the distal end, and the dome-shaped member comprising a sacrificial material.
Matrix-forming-compound deposition coating the crown and dome surface
A fabricated cornea comprising a matrix-forming compound deposited on the frame and the dome-shaped member such that the crown and at least a portion of the surface of the dome-shaped member are coated with the matrix-forming compound.
Gelatin sacrificial material for the dome-shaped member
The sacrificial material comprises gelatin.
3D-printable gelatin as sacrificial material
The gelatin comprises 3D-printable gelatin.
Insert holding assembly with well-plate manifold and wells
An assembly for holding the cell culture insert, comprising a well-plate manifold and a well-plate assembly including a plurality of wells sized to accommodate the at least one cell culture insert.
Fluid inlet and outlet lines for wells
A fluid inlet line configured to deliver fluid to the base at the proximal end of each cell culture insert and a fluid outlet line configured to remove fluid from the plurality of wells.
Matrix-forming compound selected from enumerated hydrogel, biopolymer, and synthetic polymer materials
The matrix-forming compound is chosen from a specified group comprising functionalized polyethylene glycol hydrogels, chitosan, silk, corneal extracellular matrix, elastin, fibrin, hyaluronan, collagen 2, collagen 3, collagen 4, collagen 10, mouse sarcoma cell extracellular matrix, laminin, polycaprolactone, polyethylene, polypropylene, polystyrene, nylon, polyethylene glycol and its derivatives, poly(lactic acid), poly(glycolic acid), poly(lactic acid-co-glycolic acid), polyethylene terephthalate, and combinations thereof.
Overall claim coverage centers on a cylindrical cell culture insert with a porous membrane and base, combined with a frame and a sacrificial dome-shaped member whose crown protrudes beyond the insert, followed by depositing a matrix-forming compound so the crown and dome surface are coated. The dependent claims refine the sacrificial dome material using gelatin and 3D-printable gelatin, add well-plate manifold and fluid handling features, and expand the matrix-forming compound by enumerating specific hydrogel, biopolymer, and synthetic polymer candidates.
Stated Advantages
The toroidal/annular frame supports correct geometry of the fabricated cornea scaffold.
The sacrificial dome enables shaping of the cornea without obstructing perfusion via the insert/porous membrane.
Proximal gripping/orientation is facilitated without contacting the fragile cornea.
Documented Applications
Fabricating a living cornea (corneal scaffold) using a cylindrical cell culture insert with a porous membrane, a frame, and a sacrificial dome-shaped member coated with a matrix-forming compound.
Perfusing the fabricated cornea via the insert/porous membrane to provide nutrient/oxygen transport and support matrix alignment.
Interested in licensing this patent?