Induced pluripotent stem cell (IPSC)-derived multi-ribbon biodegradable gel for outer retinal replacement

Abstract

Described herein is a method for replacing retinal tissue with a retinal cell-embedded biodegradable matrix ribbon. The biodegradable matrix ribbon can be provided as a sheet and then cut into ribbons. Alternatively, the biodegradable matrix ribbon can be constructed in ribbon form. The thin ribbon can then be implanted into a patient's eye through a small scleral and / or retinal incision. Once within the intraocular space, the ribbon can be positioned in intimate contact with the target retinal space to provide a region of biodegradable matrix with retinal cells embedded within and on its outer surface.

Description

[Technical Field]

[0001] The present invention relates to induced pluripotent stem cell (IPSC)-derived multi-ribbon biodegradable gels for outer retinal replacement. [Background technology]

[0002] Retinal degenerative diseases are one of the leading causes of irreversible blindness worldwide, with limited treatment options. Common to all retinal degenerative diseases is damage to or loss of retinal photoreceptors. Photoreceptors are light-sensing cells in the retina, the delicate layer covering the back of the eye. Photoreceptor loss can occur as a result of detachment from the underlying retinal pigment epithelium (RPE) and / or apoptosis. In either case, photoreceptor loss leads to progressive visual impairment.

[0003] One of the most common retinal degenerative diseases is age-related macular degeneration (AMD). AMD is the deterioration of the macula, the central part of the retina responsible for central, high-resolution, and color vision. AMD leads to a significant decrease in visual acuity and is the leading cause of vision loss in people over the age of 60.

[0004] Most current treatments can slow the progression of the disease but generally fail to completely halt photoreceptor cell loss. These treatments include antioxidant supplementation, intravitreal complement inhibition, lifestyle and dietary modifications, intravitreal antiangiogenic therapy, gene therapy, and implantable visual aids. Over the past decade, human pluripotent stem cells, such as embryonic stem cells and induced pluripotent stem cells, have emerged as promising options for the treatment of retinal degenerative diseases such as AMD. Embryonic stem cells (ESCs) are pluripotent stem cells derived from the inner cell mass of early preimplantation embryos. In contrast, induced pluripotent stem cells (iPSCs) do not originate from embryos but rather are derived from differentiated somatic cells. Pluripotent stem cells can self-renew and give rise to all cell lineages throughout the body, making them suitable for a wide range of applications.

[0005] Attempts have been made to address retinal degenerative diseases by transplanting induced pluripotent stem cells into the eye. However, these conventional treatment methods have certain drawbacks. For example, methods involving the injection of iPS cell suspensions suffer from low engraftment rates. Furthermore, the injection of the suspension is known to leak onto the anterior retinal surface, leading to retinal surface scarring and retinal detachment. Other methods involve the use of scaffolds to secure iPS cells in place and prevent cell loss. However, many scaffold materials, including natural or naturally derived scaffold materials such as silk, alginate, polyester, and parylene, cause chronic inflammation. Summary of the Invention [Problem to be solved by the invention]

[0006] Thus, there remains a need for methods of treating retinal degenerative diseases that can induce new cell growth and potentially reverse cell loss. [Means for solving the problem]

[0007] The present disclosure relates generally to compositions, devices and methods for replacing retinal tissue with a biodegradable matrix embedded with retinal cells.

[0008] In certain embodiments, a method of transplanting retinal cells into a patient's eye is provided, the method including combining retinal cells with a biodegradable polymer scaffold to form a gel, aspirating at least a portion of the gel into a cannula, forming an incision in the sclera of the patient's eye, inserting the cannula through the incision in the sclera of the eye, and depositing the gel in a target area of ​​the patient's eye.

[0009] In one embodiment, a method for producing a gelatinous biodegradable ribbon for retinal replacement is provided, comprising providing a container comprising a space and at least one divider member, delivering a solution of a biodegradable polymer into the space, delivering retinal cells into the space, and culturing the retinal cells and the biodegradable polymer in vitro for a period of time to provide a gelatinous ribbon comprising a biodegradable polymer matrix and the retinal cells.

[0010] In one embodiment, a method of transplanting retinal cells into a patient's eye is provided, the method including aspirating a gelatinous biodegradable ribbon into a cannula, the gelatinous biodegradable ribbon including a biodegradable polymer matrix and retinal cells, forming an incision in the sclera of the patient's eye, inserting a cannula through the incision in the sclera of the eye, and depositing the gelatinous biodegradable ribbon into a target area of ​​the patient's eye. [Brief explanation of the drawings]

[0011] [Figure 1] FIG. 1 is a schematic diagram of a culture tray for culturing retinal cells within a biodegradable polymer scaffold, according to some embodiments herein. [Figure 2] FIG. 1 is a schematic diagram of a compartmentalized culture tray for culturing retinal cells within compartmentalized ribbons of a biodegradable polymer scaffold, according to some embodiments herein. [Figure 3] FIG. 1 is a schematic diagram of a retinal cell-embedded biodegradable polymer ribbon including a visual indicator for identifying the ribbon ends, according to some embodiments herein. [Figure 4] 1 is a schematic cross-sectional view of an applicator device having a cannula with a rectangular cross-sectional area, according to some embodiments herein. [Figure 5] FIG. 10 is a schematic diagram of a proportional pedal controller that can be used with a cannula to aspirate and deposit retinal cell-embedded biodegradable polymer matrix ribbons, according to some embodiments herein. [Figure 6A]1A-1C are schematic illustrations of various operations of a method for performing retinal cell replacement, according to some embodiments herein. [Figure 6B] 1A-1C are schematic illustrations of various operations of a method for performing retinal cell replacement, according to some embodiments herein. [Figure 6C] 1A-1C are schematic illustrations of various operations of a method for performing retinal cell replacement, according to some embodiments herein. [Figure 6D] 1A-1C are schematic illustrations of various operations of a method for performing retinal cell replacement, according to some embodiments herein. [Figure 6E] 1A-1C are schematic illustrations of various operations of a method for performing retinal cell replacement, according to some embodiments herein. [Figure 6F] 1A-1C are schematic illustrations of various operations of a method for performing retinal cell replacement, according to some embodiments herein. [Figure 6G] 1A-1C are schematic illustrations of various operations of a method for performing retinal cell replacement, according to some embodiments herein. DETAILED DESCRIPTION OF THE INVENTION

[0012] To facilitate understanding, the same reference numerals have been used, wherever possible, to designate identical elements common to the figures. It is contemplated that elements and features of one embodiment may be beneficially incorporated in other embodiments without further recitation.

[0013] The retina is the light-sensitive component of the eye, transmitting information via the optic nerve to the occipital cortex of the brain. The retina is composed of layers with an ordered structure. The outer photoreceptor layer is responsible for light perception and phototransduction. The retinal pigment epithelium (RPE) layer is located between the photoreceptor layer and the underlying five-layered Bruch's membrane. The RPE layer, together with the photoreceptor layer, acts as a functional unit, and its optimal function is crucial for vision. Loss of integrity of this unit promotes photoreceptor degeneration.

[0014] Retinal degeneration occurs in several forms, including retinitis pigmentosa, macular dystrophy, and a non-neovascular form of age-related macular degeneration called geographic atrophy. Age-related macular degeneration is associated with degeneration of photoreceptor cells, while the inner retina remains intact. This multifactorial physiology makes photoreceptor replacement an effective treatment.

[0015] Surgical options for treating retinal degeneration can replace damaged retinal layers, replace lost or damaged retinal components, and / or reestablish interactions between retinal pigment epithelial cells and photoreceptors. Embryonic stem cells and existing pluripotent stem cell lines have been used as cell sources for retinal replacement, but these cells require temporary immunosuppression in patients. Furthermore, retinal pigment epithelial cell suspensions have typically been used to replace damaged retinal layers. However, these methods often face poor engraftment. Furthermore, the size of the sclerotomy is typically approximately 3.5 mm, and the transplanted cell suspension may leak through this relatively large incision. Large retinal incisions can also lead to bleeding and retinal detachment. Retinal replacement surgery has also been attempted using scaffolds to prevent cell leakage, but most biocompatible scaffold materials are non-biodegradable and cause chronic inflammation.

[0016] The embodiments described herein provide compositions and methods that address the above-mentioned problems. Such compositions and methods can be utilized to treat retinal degenerative diseases, such as age-related macular degeneration. The methods involve depositing scaffold-embedded retinal cells (e.g., polarized RPE cells and polarized photoreceptor progenitor cells) in the form of ribbons (i.e., strips). The ribbons can be provided in the form of semi-rigid and flexible gels. Multiple ribbons can be deposited adjacently on the patient's retinal surface to assemble a continuous plate-like region of cells supported by the scaffold.

[0017] Similar to straight-laid wood flooring or masonry patterns (e.g., tiling or brick paving), the ribbons can be laid side-by-side, with one ribbon length abutting and contiguous with an adjacent ribbon length. The scaffolding within each ribbon provides a framework within which and on which cells, and optionally cell nutrients and / or pharmacological agents, can be supported.

[0018] The method can include using induced pluripotent stem cells as a source of replacement retinal cells. Induced pluripotent stem cells can be prepared in a relatively short period of time (e.g., 100 days), and the patient does not need to be immunosuppressed. Replacement retinal cells derived from induced pluripotent stem cells can be provided on a biodegradable scaffold. The use of a biodegradable scaffold avoids the chronic inflammation problem that occurs when non-biodegradable scaffolds are used. The use of a scaffold material also helps maintain the cells in the desired location and prevents leakage from the eye.

[0019] Furthermore, the cell-loaded biodegradable scaffold can be provided in the form of multiple thin ribbons (i.e., strips). Because thin ribbons are used, the size of the scleral incision required to pass the cell-loaded scaffold through the ocular wall is significantly smaller than the conventional 3.5 mm (millimeter) sclerotic incision size. For example, in certain embodiments, a sclerotic incision having a maximum dimension of less than 1 mm, corresponding to a 23-gauge or smaller than 25-gauge cannula, can be used to introduce the ribbon into the eye. Similarly, in instances where the target area is within the subretinal space, the size of the retinotomy required to pass the scaffold into the subretinal space is reduced. The relatively small size of the sclerotic incision and / or retinotomy enabled by such ribbons reduces the risk of low intraocular pressure and retinal cell reflux to the anterior surface of the retina.

[0020] Multiple thin ribbons can be inserted into the eye and assembled at a target area within the eye. Multiple ribbons can be deposited adjacently under the retina to assemble multiple continuous strip areas of cells supported by the scaffold. The ribbons can be assembled in close contact, with one strip length abutting and contiguous with an adjacent strip length, similar to a straight-laid wood flooring or masonry pattern. The scaffold within each strip provides a framework within which and on which cells, and optionally cell nutrients, pharmacological agents, and / or visual indicators, can be supported. In situ assembly of multiple thin ribbons allows for the deposition of retinal cells into large, modular areas. The additional coverage provided by the larger area accommodates greater improvement in central vision without creating large retinotomy or sclerotomy.

[0021] Accordingly, some embodiments of the present disclosure are directed to a method for transplanting retinal cells into a patient's eye. In some embodiments, the method includes combining retinal cells and a biodegradable polymer scaffold to form a gel, aspirating at least a portion of the gel into a cannula of an applicator device, forming an incision in the sclera of the patient's eye, inserting the cannula of the applicator device through the incision in the sclera of the eye, and depositing the gel at a target area of ​​the patient's eye. In some embodiments, the target area includes the retina. In some embodiments, the method further includes combining at least one pharmacological agent, at least one cell nutrient, at least one visual indicator, or a combination thereof, with the retinal cells and the biodegradable polymer scaffold. In some embodiments, the target area is the subretinal space of the patient's eye. In such embodiments, the method further includes forming one or more incisions in the retina of the patient's eye, gradually inserting a cannula through each of the one or more incisions in the retina of the eye, and depositing multiple gel portions at the target area of ​​the patient's eye. In some embodiments, the patient has been diagnosed with a retinal degenerative disease and / or is in need of a retinal transplant or replacement.

[0022] Some embodiments of the present disclosure are directed to methods for producing gelatinous biodegradable ribbons for retinal replacement. In some embodiments, the methods include providing a container including a space and at least one divider member, delivering a solution of a biodegradable polymer into the space, delivering retinal cells into the space, and culturing the retinal cells and the biodegradable polymer in vitro for a period of time to form a gelatinous ribbon comprising a biodegradable polymer matrix and retinal cells. In some embodiments, the methods for producing gelatinous biodegradable ribbons for retinal replacement further include delivering at least one pharmacological agent, at least one cell nutrient, at least one visual indicator, or a combination thereof, to the space. In some embodiments, the ribbon further comprises at least one pharmacological agent, at least one cell nutrient, at least one visual indicator, or a combination thereof.

[0023] Some embodiments of the present disclosure are directed to methods of transplanting retinal cells into a patient's eye, the retinal cells being provided within a gelatinous biodegradable ribbon. In some embodiments, the method includes aspirating a gelatinous biodegradable ribbon into a cannula of an applicator device, the gelatinous biodegradable ribbon comprising a biodegradable polymer matrix and retinal cells; forming an incision in the sclera of the patient's eye; inserting the cannula of the applicator device through the incision in the sclera of the eye; and depositing the gelatinous biodegradable ribbon at a target area of ​​the patient's eye. In some embodiments, the target area of ​​the patient's eye is the subretinal space of the patient's eye. In such embodiments, the method further includes forming one or more incisions in the retina of the patient's eye, forming a subretinal bleb, inserting and positioning a cannula behind the bleb, and depositing multiple ribbons at the target area of ​​the patient's eye as the cannula is withdrawn from the bleb. In some embodiments, the ribbons are deposited in close contact with the target area of ​​the patient's eye.

[0024] Referring to FIG. 1 , a culture tray 110 and a biodegradable polymer for culturing retinal cells are shown. Within the interior volume of the culture tray 110 is a matrix 120 comprising the biodegradable polymer and retinal cells. The biodegradable polymer functions as a scaffold, providing a framework within which and on which the retinal cells, and optionally cell nutrients and / or pharmacological agents, can be supported. The matrix 120 is cut into a plurality of ribbons 130. One of the plurality of ribbons 130 has been removed from the culture tray 110, leaving an empty space corresponding to the empty tray volume 140.

[0025] In some embodiments, the biodegradable polymer is poly(lactic-co-glycolic acid) (PLGA), collagen, gelatin, the polycation poly(allyl anion hydrochloride) (PAH), the polyanion polyacrylic acid (PAA), the polycation poly(styrene sulfonic acid) (PSS), polyglycolide, poly(glycolide-co-caprolactone), poly(glycolide-co-trimethylene carbonate), polycaprolactone (PCL), polyurethane (PU), polypropylene carbonate, polyglycolic acid, The biodegradable polymer may comprise a material selected from the group consisting of polyhydroxybutyrate, polylactic acid, polydioxanone, chitosan, laminin, glycosaminoglycans, proteoglycans, heparin, elastin, fibrin, fibronectin, chondroitin sulfate proteoglycans, thiolated collagen, thiolated laminin, thiolated fibronectin, thiolated heparin, thiolated hyaluronic acid, thiolated hyaluronan-collagen-fibronectin, cellulose, hydroxyapatite, calcium phosphate, and combinations thereof. In some embodiments, the biodegradable polymer comprises a transparent or translucent material. In some embodiments, the biodegradable polymer comprises a colored material.

[0026] In some embodiments, the retinal cells comprise differentiated cells, progenitor cells, precursor cells, or a combination thereof. The differentiated cells, progenitor cells, precursor cells, or a combination thereof may comprise retinal pigment epithelial (RPE) cells, rod cells, cone cells, horizontal cells, bipolar cells, amacrine cells, retinal ganglion cells, or a combination thereof. In some embodiments, the retinal cells comprise differentiated retinal pigment epithelial cells. In some embodiments, the retinal cells comprise photoreceptor precursor cells. In some embodiments, the photoreceptor precursor cells comprise rod precursor cells, cone precursor cells, or a combination thereof. In some embodiments, the retinal cells comprise a combination of differentiated retinal pigment epithelial cells, rod precursor cells, and cone precursor cells.

[0027] In some embodiments, matrix 120, and thus each ribbon 130, comprises a monolayer of differentiated cells, such as a monolayer of differentiated RPE cells. In some embodiments, matrix 120 and each ribbon 130 comprise a bilayer of differentiated cells, such as a bilayer of differentiated RPE cells. In some embodiments, in addition to a monolayer or bilayer of differentiated cells, matrix 120 and each ribbon 130 comprise a monolayer of progenitor cells, such as a monolayer of retinal photoreceptor progenitor cells. In such embodiments, the differentiated cells may have a basal polarity, and the progenitor cells may have an apical polarity. Differentiated RPE cells, photoreceptors, and their progenitor and precursor cells may be transplanted and positioned within the patient's target retinal space using any suitable apical-basal polarity.

[0028] FIG. 2 illustrates a compartmented container 200 according to certain embodiments of the present disclosure. The compartmented container 200 includes a space or volume 210 within which a biodegradable polymer and retinal cells can be combined. Within the container space 210 are one or more dividers 220 that divide the container space into multiple sub-spaces 230. In some embodiments, the one or more dividers 220 include substantially vertical walls. In some embodiments, the sub-spaces 230 formed by the dividers 220 include elongated cross-sectional areas (e.g., shapes). In some embodiments, the sub-spaces 230 include substantially rectangular cross-sectional areas. In some embodiments, the sub-spaces 230 are cuboid-shaped, having a rectangular shape when viewed from above such that the length dimension of the sub-space is greater than the width dimension of the sub-space.

[0029] In some embodiments, a container 200 having an integer number n of partitions 220 includes an integer number n+1 of subspaces. For example, in the embodiment shown in FIG. 2, the container 200 includes 3(n) partitions 220 and 4(n+1) subspaces 230. The container 200, container space 210, partitions 220, and subspaces 230 are designed to accommodate a combination of a biodegradable polymer and retinal cells. The container 200 can accommodate the combination of a biodegradable polymer and retinal cells during a culture period to grow and proliferate the retinal cells. Cell nutrients, pharmacological agents, and / or visual indicators can be added to the combination of a biodegradable polymer and retinal cells in the container 200. Combining the biodegradable polymer and retinal cells results in a gelatinous ribbon containing retinal cells embedded within and on the biodegradable polymer, which serves as a scaffold for cell support. For example, visual indicators can be added to the ribbon to indicate the location of the ribbon's ends. The visual indicators can assist the practitioner in identifying the terminal ribbon position when positioning the ribbon during surgery. The visual indicator can be a physical characteristic of the ribbon or any dye or pigment known to those skilled in the art. An exemplary, non-limiting visual indicator is sodium fluorescein.

[0030] FIG. 3 illustrates an exemplary ribbon 300 comprising a biodegradable polymer and retinal cells, according to certain embodiments of the present disclosure. The biodegradable polymer provides a semi-rigid framework and serves as a scaffold to support the retinal cells. The ribbon 300 has a height h, a width w, and a length l. The height h, width w, and / or length l can be about 0.1 mm to about 1 mm, e.g., about 0.2 mm to about 0.8 mm, e.g., about 0.3 mm to about 0.7 mm, e.g., about 0.4 mm to about 0.6 mm, e.g., about 0.5 mm. In certain embodiments, the height h, width w, and / or length l are about 0.1 mm to about 0.9 mm, about 0.1 mm to about 0.8 mm, about 0.1 mm to about 0.7 mm, about 0.1 mm to about 0.6 mm, about 0.1 mm to about 0.5 mm, about 0.1 mm to about 0.4 mm, about 0.1 mm to about 0.3 mm, or about 0.1 mm to about 0.2 mm. In some embodiments, the height h, width w, and / or length l is from about 0.2 mm to about 1 mm, from about 0.2 mm to about 0.9 mm, from about 0.2 mm to about 0.7 mm, from about 0.2 mm to about 0.6 mm, from about 0.2 mm to about 0.5 mm, from about 0.2 mm to about 0.4 mm, or from about 0.2 mm to about 0.3 mm.

[0031] In some embodiments, the height h, width w, and / or length l are about 0.3 mm to about 1 mm, about 0.3 mm to about 0.9 mm, about 0.3 mm to about 0.8 mm, about 0.3 mm to about 0.6 mm, about 0.3 mm to about 0.5 mm, or about 0.3 mm to about 0.4 mm. In some embodiments, the height h, width w, and / or length l are about 0.4 mm to about 1 mm, about 0.4 mm to about 0.9 mm, about 0.4 mm to about 0.8 mm, about 0.4 mm to about 0.7 mm, or about 0.4 mm to about 0.5 mm. In some embodiments, the height h, width w, and / or length l are about 0.5 mm to about 1 mm, about 0.5 mm to about 0.9 mm, about 0.5 mm to about 0.8 mm, about 0.5 mm to about 0.7 mm, or about 0.5 mm to about 0.6 mm.

[0032] In some embodiments, the height h, width w, and / or length l are about 0.6 mm to about 1 mm, about 0.6 mm to about 0.9 mm, about 0.6 mm to about 0.8 mm, or about 0.6 mm to about 0.7 mm. In some embodiments, the height h, width w, and / or length l are about 0.7 mm to about 1 mm, about 0.7 mm to about 0.9 mm, or about 0.7 mm to about 0.8 mm. In some embodiments, the height h, width w, and / or length l are about 0.8 mm to about 1 mm, or about 0.8 mm to about 0.9 mm. In some embodiments, the height h, width w, and / or length l are about 0.9 mm to about 1 mm.

[0033] In some embodiments, ribbon 300 has dimensions based on the size of the incision in the sclera and / or retina of the eye. For example, in some embodiments, ribbon 300 has lateral dimensions that facilitate a small scleral incision for introducing ribbon 300 into the eye. In some embodiments, multiple ribbons 300 applied to a target area have overall dimensions equal to or substantially equal to the size of the visual field to be restored.

[0034] The exemplary ribbon 300 shown in FIG. 3 has a rectangular parallelepiped shape with length l greater than width w and rectangular faces (e.g., rectangular cross-section) when viewed from above, although other shapes are contemplated, including three-dimensional shapes with square, diamond, rounded, and triangular faces, for example. Ribbon 300 includes optional visual indicator 310 and optional visual indicator 320 located near the ribbon ends. Visual indicator 310 and visual indicator 320 can assist a practitioner in identifying the ribbon ends and positioning ribbon 300 during surgery. In some embodiments, multiple ribbons 300 placed in a patient's eye can be equal in size or different in size. In some embodiments, visual indicator 310 includes a physical feature, such as a hole, indentation, fiducial marking, or the like, formed in ribbon 300. In some embodiments, visual indicator 310 includes a dye or pigment disposed at or near the ribbon ends.

[0035] FIG. 4 illustrates an applicator 400 according to some embodiments. The applicator 400 can be used to deliver and apply one or more ribbons of biodegradable polymer and retinal cells to a target area within a patient's eye. Generally, the applicator 400 includes a handle 450 and a cannula 410. The cannula 410 has a proximal end 412 coupled to a distal end 454 of the handle 450 and a distal end 414 opposite the proximal end 412. In some embodiments, a flexible tube 460 can be disposed through or coupled to the proximal end 452 of the handle 450 to provide vacuum suction to the cannula 410 for aspirating the ribbons. The flexible tube 460 can be in fluid communication with a vacuum source at its proximal end and in direct or indirect fluid communication with the cannula 410 at its distal end 464. For example, flexible tubing 460 may be directly coupled to cannula 410 through handle 450, or flexible tubing 460 may be indirectly coupled to cannula 410 via interior chamber 456 of handle 450. Activation of the vacuum source, and therefore the delivery of vacuum through flexible tubing 460, may be controlled by a proportional foot pedal or a toggle on applicator 400.

[0036] The cannula 410, which may include a tube, is generally formed of any suitable surgical-grade material, such as a metal or a thermoplastic polymer material. Examples of metal materials include aluminum, stainless steel, and other metal alloys. Examples of suitable thermoplastic polymer materials include polyetheretherketone (PEEK), polyetherketone (PEK), and polytetrafluoroethylene (PTFE). In some embodiments, the interior surface of the cannula 410 comprises a surface material selected from the group consisting of fluorinated ethylene propylene, polytetrafluoroethylene, and the like.

[0037] In some embodiments, cannula 410 comprises a rectangular-shaped tube (e.g., having a rectangular cross-section), and / or the tube of cannula 410 comprises a rectangular-shaped distal tip. In some embodiments, cannula 410 has a rectangular-shaped opening 420 at its distal end 414. The rectangular shape of cannula 410 and / or opening 420 may facilitate aspiration and transfer of ribbons having a rectangular cross-sectional area. The rectangular cross-section ribbons disclosed herein can also be aspiration and transfer using conventional cannulas having a circular cross-section.

[0038] In some embodiments, applicator 400 further includes a plunger 470 disposed through at least a portion of handle 450 (e.g., a portion of internal chamber 456) and configured to translate into and through cannula 410. Plunger 470 can be used to push or eject a ribbon drawn into cannula 410 to apply the ribbon to a target area of ​​the eye during a surgical procedure. In some embodiments, plunger 470 is attached to a sliding toggle 472 movably coupled to handle 450, facilitating manual control of plunger 470 by a user. However, in other embodiments, plunger 470 can be electromechanically or pneumatically actuated, for example, by an electromechanical motor or a supply of fluid pressure via flexible tubing 460. In such embodiments, plunger 470 can be controlled by a foot pedal or other user input device.

[0039] FIG. 5 illustrates a proportional pedal controller 500 that can be used with an applicator device, such as applicator 400, for aspirating and depositing ribbons as disclosed herein. The proportional pedal controller 500 includes a foot pedal 510 hinged at a fulcrum 520. The pedal 510 is coupled to a sensing arm 530 that is connected to a controller unit 540. The proportional pedal controller 500 can be communicatively coupled to the applicator device, and the proportional pedal controller and the applicator device can be used together to aspirate one or more ribbons into a cannula of the applicator device and deposit the ribbons from the cannula into an appropriate location within a patient's eye. The foot pedal 510 can be coupled to the applicator device via a variety of coupling means, including a hydraulic coupling, an electronic sensor, a computerized coupling using one or more servo motors, a direct pressure transmission coupling, or the like.

[0040] 6A-6F illustrate various operations during an exemplary method of retinal cell replacement, in which a target region is positioned within or adjacent to the subretinal space. As shown in FIG. 6A, a biodegradable polymer scaffold sheet 610 is embedded with a monolayer or bilayer of retinal pigment epithelial (RPE) cells 612 and retinal photoreceptor progenitor cells 614, which may be derived from induced pluripotent stem cells. The cells are arranged with the RPE cells 612 having a basal polarity and the retinal photoreceptor progenitor cells 614 having an apical polarity. In FIG. 6B, the sheet is cut into multiple ribbons 616, and in FIG. 6C, the ribbons 616 are drawn into a cannula 632 of a ribbon applicator device (e.g., applicator 400) by applying vacuum suction through the cannula 632. In some embodiments, visual indicators included on the ribbon ends assist the user in identifying where the ribbons 616 begin and end.

[0041] In Figure 6D, scissors or other surgical instrument 622 are introduced into the patient's eye 620 (e.g., via a trocar cannula and sclerotomy) and guided through the intraocular space 644 thereof to create an incision 624 (e.g., a retinotomy) in the retina 626 of the eye 620. In Figure 6E, an injection device 628 is introduced into the eye 620 and a buffered salt solution (e.g., BSS) or viscoelastic is injected through the incision 624 into the subretinal space 640 to form a bleb 630. In Figure 6F, after removal of the injection device 628, a cannula 632 of a ribbon applicator device is introduced into the eye 620 and guided through the incision 624 to the back of the bleb 630. Then, in FIG. 6G, as cannula 632 is slowly withdrawn from bleb 630, multiple ribbons 616 within cannula 632 are injected (eg, by actuation of a plunger or other injection mechanism) from cannula 632 onto target area 642.

[0042] In some embodiments, the ribbons 616 are closely deposited on the target area 642 of the patient's eye. The closely deposited ribbons 616 can resemble straight wood flooring or a masonry pattern (e.g., tiling or brick paving), where the ribbons 616 are deposited side-by-side such that the ribbons 616 abut and are continuous with adjacent ribbons 616.

[0043] After the cannula 632 is withdrawn from the eye 620, this process may be repeated one or more times through the same or additional retinotomy to apply additional ribbons 616. The additional ribbons 616 may be placed adjacent to the previously deposited ribbons 616 such that the ribbons 616 are positioned in close contact.

[0044] Illustrative Embodiments Embodiment 1: A method of producing a gelatinous biodegradable ribbon for retinal replacement, comprising: providing a container comprising a space and at least one divider member; delivering a solution of a biodegradable polymer to the space; delivering retinal cells to the space; and culturing the retinal cells and the biodegradable polymer in vitro for a period of time to provide a gelatinous ribbon comprising a biodegradable polymer matrix and the retinal cells.

[0045] Embodiment 2: The method of embodiment 1, wherein at least one partition member divides the space into a first subspace and a second subspace.

[0046] Embodiment 3: The method of embodiment 1, wherein an integer number n of partition members divide the space into an integer number n+1 of subspaces.

[0047] Embodiment 4: The method of embodiment 1, wherein the container comprises a substantially vertical interior wall.

[0048] Embodiment 5: The method of embodiment 1, wherein at least one partition member comprises a substantially vertical wall.

[0049] Embodiment 6: The method of embodiment 2, wherein the first subspace and the second subspace are rectangular parallelepiped shapes in which the length dimension of the subspaces is greater than the width dimension of the subspaces.

[0050] Embodiment 7: The method of embodiment 3, wherein each of the integer n+1 subspaces is a rectangular parallelepiped shape with a length dimension of the subspace greater than a width dimension of the subspace.

[0051] Embodiment 8: The method of embodiment 2, wherein the first subspace and the second subspace each comprise a substantially rectangular cross-sectional area.

[0052] Embodiment 9: The method of embodiment 3, wherein each of the integer n+1 subvolumes comprises a substantially rectangular cross-sectional area.

[0053] Embodiment 10: The method of embodiment 1, wherein the retinal cells comprise differentiated cells, progenitor cells, precursor cells, or a combination thereof.

[0054] Embodiment 11: The method of embodiment 1, wherein the retinal cells are selected from the group consisting of retinal pigment epithelial cells, rod cells, cone cells, horizontal cells, bipolar cells, amacrine cells, retinal ganglion cells, and combinations thereof.

[0055] Embodiment 12: The method of embodiment 1, wherein the retinal cells comprise differentiated retinal pigment epithelial cells.

[0056] Embodiment 13: The method of embodiment 1, wherein the retinal cells comprise photoreceptor progenitor cells.

[0057] Embodiment 14: The method of embodiment 13, wherein the photoreceptor progenitor cells comprise rod progenitor cells, cone progenitor cells, or a combination thereof.

[0058] Embodiment 15: The method of embodiment 1, wherein the retinal cells comprise differentiated retinal pigment epithelial cells, rod progenitor cells, and cone progenitor cells.

[0059] Embodiment 16: The biodegradable polymer is poly(lactic-co-glycolic acid) (PLGA), collagen, gelatin, the polycation poly(allyl anion hydrochloride) (PAH), the polyanion polyacrylic acid (PAA), the polycation poly(styrene sulfonic acid) (PSS), polyglycolide, poly(glycolide-co-caprolactone), poly(glycolide-co-trimethylene carbonate), polycaprolactone (PCL), polyurethane (PU), polypropylene carbonate, polyglycolic acid, poly 2. The method of embodiment 1, comprising a polymer selected from the group consisting of hydroxybutyric acid, polylactic acid, polydioxanone, chitosan, laminin, glycosaminoglycans, proteoglycans, heparin, elastin, fibrin, fibronectin, chondroitin sulfate proteoglycans, thiolated collagen, thiolated laminin, thiolated fibronectin, thiolated heparin, thiolated hyaluronic acid, thiolated hyaluronan-collagen-fibronectin, cellulose, calcium phosphate, and combinations thereof.

[0060] Embodiment 17: The method of embodiment 1, further comprising delivering at least one pharmacological agent, at least one cellular nutrient, or a combination thereof, to the space.

[0061] Embodiment 18: A method for transplanting retinal cells into a patient's eye, the method comprising aspirating a gelatinous biodegradable ribbon into a cannula, the gelatinous biodegradable ribbon comprising a biodegradable polymer matrix and retinal cells; forming an incision in the sclera of the patient's eye; inserting a cannula through the incision in the sclera of the eye; and depositing the gelatinous biodegradable ribbon into a target area of ​​the patient's eye.

[0062] Embodiment 19: The method of embodiment 18, wherein the retinal cells comprise differentiated cells, progenitor cells, precursor cells, or a combination thereof.

[0063] Embodiment 20: The method of embodiment 19, wherein the differentiated cells, progenitor cells, precursor cells, or combinations thereof comprise retinal pigment epithelial cells, rod cells, cone cells, horizontal cells, bipolar cells, amacrine cells, retinal ganglion cells, or combinations thereof.

[0064] Embodiment 21: The method of embodiment 18, wherein the retinal cells comprise differentiated retinal pigment epithelial cells.

[0065] Embodiment 22: The method of embodiment 18, wherein the retinal cells comprise photoreceptor progenitor cells.

[0066] Embodiment 23: The method of embodiment 22, wherein the photoreceptor progenitor cells are selected from the group consisting of rod progenitor cells, cone progenitor cells, and combinations thereof.

[0067] Embodiment 24: The method of embodiment 18, wherein the retinal cells comprise differentiated retinal pigment epithelial cells, rod progenitor cells, and cone progenitor cells.

[0068] Embodiment 25: The method of embodiment 18, further comprising depositing a bilayer of ribbons by aspirating a second gelatinous biodegradable ribbon into the cannula or a second cannula, the second gelatinous biodegradable ribbon comprising a second biodegradable polymer matrix and additional retinal cells, inserting the cannula or the second cannula through an incision in the sclera of the eye, and depositing the second gelatinous biodegradable ribbon at the target area or a second target area of ​​the patient's eye.

[0069] Embodiment 26: The method of embodiment 25, wherein the bilayer of ribbons comprises (1) a first basal ribbon of the bilayer of ribbons comprising differentiated retinal pigment epithelial cells, and (2) a second apical gel ribbon comprising photoreceptor progenitor cells.

[0070] Embodiment 27: The method of embodiment 18, further comprising forming a plurality of incisions in the retina of the patient's eye, gradually inserting a cannula through each of the plurality of incisions in the retina of the eye, and depositing a plurality of ribbons in a target area of ​​the patient's eye.

[0071] Embodiment 28: The method of embodiment 27, further comprising depositing the ribbon in close contact with the target area of ​​the patient's eye.

[0072] Embodiment 29: The biodegradable polymer matrix is ​​selected from the group consisting of poly(lactic-co-glycolic acid) (PLGA), collagen, gelatin, the polycation poly(allyl anion hydrochloride) (PAH), the polyanion (polyacrylic acid) (PAA), the polycation poly(styrene sulfonic acid) (PSS), polyglycolide, poly(glycolide-co-caprolactone), poly(glycolide-co-trimethylene carbonate), polycaprolactone (PCL), polyurethane (PU), polypropylene carbonate, polyglycolic acid, polyhydrogen 19. The method of embodiment 18, wherein the polymer is selected from the group consisting of: polybutyric acid, polylactic acid, polydioxanone, chitosan, laminin, glycosaminoglycans, proteoglycans, heparin, elastin, fibrin, fibronectin, chondroitin sulfate proteoglycans, thiolated collagen, thiolated laminin, thiolated fibronectin, thiolated heparin, thiolated hyaluronic acid, thiolated hyaluronan-collagen-fibronectin, cellulose, hydroxyapatide, calcium phosphate, and combinations thereof.

[0073] Embodiment 30: The method of embodiment 29, wherein the biodegradable polymer matrix comprises PLGA.

[0074] Embodiment 31: The method of embodiment 18, further comprising depositing the ribbon into the subretinal space of the patient's eye.

[0075] Embodiment 32: The method of embodiment 18, wherein the cannula comprises a substantially rectangular cross-sectional area.

[0076] Embodiment 33: The method of embodiment 18, wherein the cannula comprises a surface material selected from the group consisting of fluorinated ethylene propylene, polytetrafluoroethylene, and others included.

[0077] Embodiment 34: The method of embodiment 18, wherein the ribbon further comprises at least one pharmacological agent, at least one cell nutrient, or a combination thereof.

[0078] The detailed description and drawings support and explain the disclosure, the scope of which is defined only by the claims. Although some of the best modes and alternative embodiments for carrying out the claimed disclosure have been described in detail, there are various alternative designs and embodiments for carrying out the disclosure as defined in the appended claims.

[0079] Furthermore, the features of the various embodiments illustrated in the drawings or described herein should not necessarily be understood as independent embodiments. Rather, each of the characteristics described in one example embodiment may be combined with one or more other desirable characteristics from other embodiments, resulting in other embodiments not described in words or by reference to drawings. Accordingly, such other embodiments may be included within the scope of the appended claims.

Claims

1. 1. A method for transplanting retinal cells into an eye of a patient, comprising: combining retinal cells with a biodegradable polymer scaffold to form a gel; aspirating at least a portion of the gel into a first cannula; forming an incision in the sclera of the patient's eye; inserting the first cannula through the incision in the sclera of the patient's eye; depositing the gel onto a first target area of ​​the patient's eye; A method comprising:

2. 2. The method of claim 1, wherein the retinal cells are selected from the group consisting of differentiated cells, progenitor cells, precursor cells, and combinations thereof.

3. 2. The method of claim 1, wherein the retinal cells are selected from the group consisting of retinal pigment epithelial cells, rod cells, cone cells, horizontal cells, bipolar cells, amacrine cells, retinal ganglion cells, and combinations thereof.

4. The method of claim 1 , wherein the retinal cells comprise differentiated retinal pigment epithelial cells.

5. The method of claim 1 , wherein the retinal cells include photoreceptor progenitor cells.

6. The method of claim 5 , wherein the photoreceptor progenitor cells include rod progenitor cells, cone progenitor cells, or a combination thereof.

7. The method of claim 1 , wherein the retinal cells comprise differentiated retinal pigment epithelial cells, rod progenitor cells, and cone progenitor cells.

8. combining additional retinal cells with the second biodegradable polymer scaffold to form a second gel; aspirating at least a portion of the second gel into the first cannula or the second cannula; inserting the first cannula or the second cannula through the incision in the sclera of the patient's eye; depositing the second gel onto the first target area or a second target area of ​​the patient's eye; 10. The method of claim 1, further comprising depositing a bilayer of gel by

9. The method of claim 1 , wherein the gel comprises a first basal layer comprising differentiated retinal pigment epithelial cells and a second apical layer comprising photoreceptor precursor cells.

10. The method of claim 1 , further comprising cutting the gel into one or more ribbons before aspirating the gel into the first cannula.

11. The method of claim 10 , wherein the one or more ribbons have a substantially rectangular cross-sectional area.

12. 11. The method of claim 10, further comprising depositing the one or more ribbons in close contact with the first target area or the second target area of ​​the patient's eye.

13. The biodegradable polymer scaffolds include poly(lactic-co-glycolic acid) (PLGA), collagen, gelatin, polycation poly(allyl anion hydrochloride) (PAH), polyanion (polyacrylic acid) (PAA), polycation poly(styrene sulfonic acid) (PSS), polyglycolide, poly(glycolide-co-caprolactone), poly(glycolide-co-trimethylene carbonate), polycaprolactone (PCL), polyurethane (PU), polypropylene carbonate, polyglycolic acid, polyhydroxybutyric acid.

10. The method of claim 1, wherein the grafted tissue comprises a material selected from the group consisting of polylactic acid, polydioxanone, chitosan, laminin, glycosaminoglycans, proteoglycans, heparin, elastin, fibrin, fibronectin, chondroitin sulfate proteoglycans, thiolated collagen, thiolated laminin, thiolated fibronectin, thiolated heparin, thiolated hyaluronic acid, thiolated hyaluronan-collagen-fibronectin, cellulose, hydroxyapatide, calcium phosphate, and combinations thereof.

14. The method of claim 13 , wherein the biodegradable polymer scaffold comprises PLGA.

15. The method of claim 1 , further comprising depositing the gel in the subretinal space of the patient's eye.

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