MACS-based purification of stem cell-derived retinal pigment epithelium
Inventors
Meyer, Nathan • Chase, Lucas • Stankewicz, Casey
Assignees
Interested in licensing this patent?
MTEC can help explore whether this patent might be available for licensing for your application.
Publication Number
US-11162070-B2
Publication Date
2021-11-02
Expiration Date
Abstract
Provided herein are methods of enriching a retinal pigment epithelium (RPE) cell population derived from stem cells. Such a method may comprise removing contaminating cells through the depletion of CD24 positive cells, CD56 positive cells, and/or CD90 positive cells from a starting population of RPE cells.
Core Innovation
The invention concerns a method for providing a human retinal pigment epithelium (RPE)-enriched cell population from a less RPE-enriched starting human RPE cell population. The method includes depleting CD56+ cells from the starting RPE cell population to provide the RPE-enriched cell population, which is characterized by high proportions of RPE marker-positive cells.
In the provided RPE-enriched cell population, at least 90% of the cells are BEST1+, at least 90% of the cells are PMEL17+, and at least 90% of the cells are TYRP1+. Additional refinements include producing RPE-enriched populations with higher marker purity, including at least 95% for BEST1+, PMEL17+, and TYRP1+.
The approach is directed to stem cell-derived RPE enrichment, including RPE-enriched cells derived from pluripotent stem cells, including ES/iPSCs. The depletion/enrichment strategy is further refined by additionally depleting contaminating CD24+ and/or CD90+ cells, and the resulting RPE-enriched cells are described as essentially non-genetically modified and cryopreservable for research/screening and therapeutic use.
Claims Coverage
The independent claim set centers on CD56+ depletion to provide a human RPE-enriched cell population with three-marker purity, requiring at least 90% BEST1+, at least 90% PMEL17+, and at least 90% TYRP1+ cells. Dependent refinements tighten purity thresholds and expand depletion targets.
CD56+ depletion to provide an RPE-enriched cell population with marker purity
Depleting CD56+ cells from the starting RPE cell population to provide the RPE-enriched cell population wherein at least 90% of the cells are BEST1+, at least 90% are PMEL17+, and at least 90% are TYRP1+.
At least 95% marker purity for BEST1+, PMEL17+, and TYRP1+
Providing an RPE-enriched cell population wherein at least 95% of the cells express BEST1+, PMEL17+, and TYRP1+.
Additional depletion of CD24+ cells
Further enriching the starting cell population for RPE-enriched cells by additionally depleting CD24+ cells.
Additional depletion of CD90+ cells
Further enriching an initial cell population for RPE cells by depleting CD90+ cells.
CD56+ depletion by magnetic-bead-based sorting or fluorescence-based sorting
Depleting CD56+ cells using magnetic-bead-based sorting or fluorescence-based sorting.
Provided RPE-enriched cell population is not genetically modified
Providing an RPE-enriched cell population that is not genetically modified.
Overall, the claims focus on CD56+ depletion to generate a human RPE-enriched cell population with high proportions of BEST1+, PMEL17+, and TYRP1+ cells. Dependent claims further tighten marker purity, add CD24+ and/or CD90+ depletion, specify CD56+ depletion modality, and restrict the population to being not genetically modified.
Stated Advantages
Enables provision of an essentially non-genetically modified, cryopreservable RPE-enriched cell population.
Increases RPE marker enrichment to high purity levels based on BEST1, PMEL17, and TYRP1.
Supports time-/cost-efficient production of RPE-enriched cells for research/screening and therapeutic use.
Documented Applications
Research and screening using cryopreservable, essentially non-genetically modified RPE-enriched cells.
Therapeutic use of cryopreservable, essentially non-genetically modified RPE-enriched cells.
Interested in licensing this patent?