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Figure 1. Workflow for the Differentiation of hPSCs into Retinal Pigment Epithelium (RPE) with ���շ��ѻ徱�ڴ�™ RPE Culture System

hPSC colonies, previously harvested and seeded as clumps, are put directly into the medium provided in ���շ��ѻ徱�ڴ�™-ACF RPE Differentiation Kit. Seeding into ���շ��ѻ徱�ڴ�™ RPE Differentiation Medium A induces cells toward immature RPE. A full medium change is performed on Day 1 with fresh ���շ��ѻ徱�ڴ�™ RPE Differentiation Medium A and then on Day 2 with ���շ��ѻ徱�ڴ�™ RPE Differentiation Medium B. On Days 4 and 6, medium changes are performed with fresh ���շ��ѻ徱�ڴ�™ RPE Differentiation Medium C. On Days 7 and every second day thereafter a medium change is performed with fresh ���շ��ѻ徱�ڴ�™ RPE Differentiation Medium D. On Day 14, immature RPE cells are enzymatically harvested and subcultured in ���շ��ѻ徱�ڴ�™-XF RPE Maturation Medium with the addition of ���շ��ѻ徱�ڴ�™-ACF RPE Plating Supplement from Days 14 to 21 to improve survival after passaging. RPE cells begin to mature over the course of 5 weeks of culture in ���շ��ѻ徱�ڴ�™-XF RPE Maturation Medium and become fully matured by Day 49, possessing key characteristics such as polygonal shape, polarization, pigmentation and phagocytosis.

Figure 2. ���շ��ѻ徱�ڴ�™ Retinal Pigment Epithelial (RPE) Plating Supplement Improves Attachment and Morphology of Replated hPSC-Derived Immature RPE Cells Derived From 4 hPSC Lines

(A) Four different cell lines of hPSC-derived immature RPE cells were replated in either ���շ��ѻ徱�ڴ�™-XF RPE Maturation Medium alone, or (B) with added ���շ��ѻ徱�ڴ�™-ACF RPE Plating Supplement (RPE Plating Medium). (A) Representative images of replated hPSC-derived immature RPE cells are shown 1 Day after replating. (B) hPSC-derived immature RPE cells exhibit improved attachment when replated with RPE Plating Medium. Data shown as mean + SEM; n = 3. (C) Plating Efficiency was measured across the four cell lines following replating showing a 3-4 fold increase with the ���շ��ѻ徱�ڴ�™ RPE-ACF Plating Supplement.

Figure 3. hPSC-Derived Immature Retinal Pigment Epithelial (RPE) Cells Replated Using ���շ��ѻ徱�ڴ�™-ACF RPE Plating Supplement Show Increased Cell Survival and Improved Morphology Post Cryopreservation

(A) Four different cell lines of cryopreserved hPSC-derived immature RPE cells were thawed and replated in either ���շ��ѻ徱�ڴ�™-XF RPE Maturation Medium alone, or (B) with ���շ��ѻ徱�ڴ�™-ACF RPE Plating Supplement (RPE Plating Medium). Representative images show cells 1 Day post-thaw. Post-thaw, hPSC-derived immature RPE cells exhibited improved attachment when replated with RPE Plating Medium. Data shown as mean + SEM; n = 3. (C) Post-Thaw Plating Efficiency was measured across the four cell lines following replating showing a 3-4 fold increase with the ���շ��ѻ徱�ڴ�™-ACF RPE Plating Supplement.