Method for differentiation of ocular cells and use thereof
Inventors
Chase, Lucas • WALLACE, Kyle • Meline, Benjamin • DIAS, Andrew • Shelley, Brandon • FENN, Marisa • PHILLIPS, Debjani • STERNFELD, Matthew • Meyer, Nathan
Assignees
Interested in licensing this patent?
MTEC can help explore whether this patent might be available for licensing for your application.
Publication Number
US-12110503-B2
Publication Date
2024-10-08
Expiration Date
Abstract
Provided herein are methods of producing a photoreceptor precursor (PRP) cell population derived from stem cells. Further provided herein are methods of using the PRP cell populations, such as for therapeutics.
Core Innovation
The invention provides a method for producing a population of neural retinal progenitors (NRPs) from human induced pluripotent stem cells (iPSCs) by performing a staged retinal differentiation using first and second retinal differentiation media (RD1 and RD2) followed by a retinal maturation (RM) medium. Cells are cultured in RD1 comprising a BMP inhibitor to further differentiate the cells to anterior neuroectoderm cells, and retinal differentiation is then induced by culturing the anterior neuroectoderm cells in RD2 essentially free of BMP inhibitors to form retinal progenitor cells (RPCs).
After RPC formation, the RPCs are cultured in an RM medium to produce NRPs. The staged differentiation sequence is presented as a workflow to generate NRPs from iPSCs through an anterior neuroectoderm and RPC intermediate, with constraints on medium components and the essentially-free nature of specific factors, including BMP inhibitors during RD2 and Activin A being essentially free in the RM medium.
The described subject matter also encompasses selecting and refining NRP populations based on marker-based enrichment and quality control concepts. NRPs are characterized using marker expression thresholds for genes and proteins such as PAX6, PMEL17, and VSX2, and a composition is included that defines an NRP population derived from iPSCs with specified minimum fractions of cells expressing these markers.
Claims Coverage
The document provides two independent claims: a method for producing NRPs and a defined NRP composition derived from iPSCs. Across these, the inventive coverage centers on a staged RD1/RD2/RM differentiation workflow with BMP-inhibitor control and on defined marker-expression thresholds in the resulting NRP population.
Staged retinal differentiation using RD1 with BMP inhibitor and RD2 essentially free of BMP inhibitors
Culturing human iPSCs in a first retinal differentiation medium (RD1) comprising a BMP inhibitor to differentiate the cells to anterior neuroectoderm cells; inducing retinal differentiation by culturing the anterior neuroectoderm cells in a second retinal differentiation medium (RD2) essentially free of BMP inhibitors to form retinal progenitor cells (RPCs).
Retinal maturation of RPCs to produce NRPs using an RM medium
Culturing the RPCs in a retinal maturation (RM) medium to produce NRPs.
Defined NRP composition derived from iPSCs with marker-expression thresholds
A composition comprising an NRP population derived from iPSCs wherein at least 90% of the cells express PAX6, at least 90% of the cells express PMEL17, and at least 70% of the cells express VSX2.
Claim coverage focuses on generating NRPs from iPSCs via RD1/RD2/RM stages that control BMP-inhibitor presence, and on specifying a final NRP composition by quantitative expression of PAX6, PMEL17, and VSX2.
Stated Advantages
Not explicitly described in patent.
Documented Applications
Therapeutic administration for retinal degeneration [procedural detail omitted for safety].
In vitro compound screening assays using the described retinal lineages and progenitor/precursor populations [procedural detail omitted for safety].
Interested in licensing this patent?