Protocol for purifying iPSC-derived retinal pigment epithelial cells by sequential digestion and intrinsic autofluorescence sorting.

Induced pluripotent stem cells (iPSC)-derived retinal pigment epithelium cells serve as a promising cell source for transplantation therapy in age-related macular degeneration. Here, we present a protocol for purifying the mature retinal pigment epithelium (RPE) cells post-differentiation. We describe steps for sequentially digesting cells with different enzymes and sorting them based on the autofluorescence of mature RPE cells, avoiding fixation and immunostaining. By maintaining high cell viability and purity, this cost-efficient protocol has broad implications for both in vitro research and transplantation applications.