Macs-based purification of stem cell-derived retinal pigment epithelium
Inventors
Meyer, Nathan • Chase, Lucas • Stankewicz, Casey
Assignees
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Publication Number
US-12241083-B2
Publication Date
2025-03-04
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 relates to a method for enriching for RPE cells from a starting population of RPE cells by depleting CD56+ cells. The method provides an RPE-enriched cell population in which at least 90% of the cells are PMEL17+ and at least 90% of the cells are TYRP1+.
The described approach further frames depletion as removing contaminating cells defined by cell-surface markers, including CD56 and optionally additional markers such as CD24 and CD90. The enrichment outcome is evaluated using flow-cytometry marker panels before and after depletion, including PMEL17, TYRP1, BEST1, CRALBP, and other associated markers.
The invention also connects the enriched RPE population to downstream characterization and use, including RPE maturation and functional characterization by tight junction markers, ultrastructure, and functional readouts such as TER/TEP. The document additionally describes cryopreservation and reproducibility across iPSC lines and operators.
Claims Coverage
The partial content includes one independent claim directed to CD56+ depletion for enriching an RPE-enriched cell population defined by PMEL17+ and TYRP1+ thresholds. The claim family summary also describes additional depletion markers such as CD90+ and optionally CD24+.
Depleting CD56+ cells to produce an RPE-enriched population
A method for enriching for RPE cells from a starting population of RPE cells by depleting CD56+ cells from the starting RPE cell population to provide the RPE-enriched cell population.
Defining RPE-enriched purity with PMEL17 and TYRP1 thresholds
The RPE-enriched cell population wherein at least 90% of the cells are PMEL17+ and at least 90% of the cells are TYRP1+.
Overall, the claim coverage centers on CD56+ depletion from a starting RPE cell population and the definition of success by PMEL17+ and TYRP1+ marker positivity thresholds in the resulting RPE-enriched cell population.
Stated Advantages
Enables enrichment of RPE cells such that at least 90% of the enriched population is PMEL17+ and TYRP1+.
Improves RPE marker positivity using depletion of contaminating cell populations expressing CD56 and optionally CD24 and/or CD90.
Supports downstream RPE maturation and functional characterization by tight junction markers, ultrastructure, and TER/TEP.
Supports cryopreservation and reproducibility across iPSC lines and operators.
Documented Applications
Screening using the enriched RPE cells as described in the document.
Therapeutic transplantation using the enriched RPE cells as described in the document.
Cryopreservation of the enriched RPE cells and use across iPSC lines and operators as described in the document.
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