The nuclear density of hES-RPE grown in BCEC-CM (mean, 20.1 nuclei/mm Bruch's membrane) was significantly higher than that of cells grown in CM vehicle (mean, 5.1 nuclei/mm Bruch's membrane) (
P = 0.031). Fetal RPE cell nuclear density after culture in BCEC-CM (mean, 20.6 nuclei/mm Bruch's membrane) was significantly higher than that of cells cultured in CM vehicle (mean, 2.2 nuclei/mm Bruch's membrane) (
P < 0.001). The nuclear density of adult RPE cells cultured in BCEC-CM (mean, 10.0 nuclei/mm Bruch's membrane) was also significantly higher than the nuclear density of cells cultured in CM vehicle (mean, 1.2 nuclei/mm Bruch's membrane) (
P = 0.016) (see
Fig. 6A). ANOVA on ranks showed significant differences in the median values among the cell types cultured in BCEC-CM (
P = 0.004). The nuclear densities of fetal RPE and hES-RPE cultured in BCEC-CM were significantly higher than the nuclear density of adult RPE cultured in BCEC-CM (
P < 0.05). Fetal RPE and hES-RPE nuclear densities were not significantly different when cultured in BCEC-CM (
P > 0.05). Nuclear densities of hES-RPE, fetal RPE, and cultured adult RPE were not significantly different when cultured in CM vehicle only (
P = 0.060). There were no statistically significant differences in ages of donor eye explants between groups (
P = 0.345: hES-RPE mean donor explant age, 80.2 ± 8.4 years; fetal RPE mean donor explant age, 80.2 ± 7.8 years; aged adult RPE mean donor explant age, 75.7 ± 3.6 years). We assessed whether RPE survival on age-matched AMD versus non-AMD Bruch's membrane was similar in the presence of BCEC-CM (
Fig. 6B). Explants seeded with fetal RPE on aged Bruch's membrane from eyes without significant AMD changes (including donor eyes with submacular focal RPE hyperplasia and few small [<100 μm] drusen) were compared with explants seeded on AMD Bruch's membrane. AMD donor eyes included those with CNV (removed before cell seeding with no subsequent debridement), geographic atrophy, and extensive large (≥125 μm) drusen. After culture in BCEC-CM, non-AMD donor eye mean nuclear density was 19.6 nuclei/mm Bruch's membrane, and AMD donor eye mean nuclear density was 21.3 nuclei/mm Bruch's membrane. After culture in CM vehicle, non-AMD donor eye mean nuclear density was 2.4 nuclei/mm Bruch's membrane, and AMD donor eye mean nuclear density was 2.0 nuclei/mm Bruch's membrane. Differences in fetal RPE nuclear density on Bruch's membrane in the presence of BCEC-CM versus CM vehicle were statistically significant for both non-AMD and AMD donors (non-AMD,
P = 0.004; AMD,
P < 0.001). For a given culture medium, the nuclear densities of fetal RPE on non-AMD versus AMD explants were not significantly different (culture in BCEC-CM,
P = 0.548; culture in CM vehicle,
P = 0.231). Ages of the two groups were not statistically different (
P = 0.226: aged, non-AMD mean donor age, 77.8 ± 5.0 years; AMD mean donor age, 82.2 ± 9.3 years).