At day 21 from the 20 start of the floating cu1ture, the กลุ่มก้อนs were disrupted by injecting and ~jecting solution by micropipette, and the resulting cells were subjected to การเพาะเลี้ยงแบบยึดเกาะ in a serum• free อาหาร, which was DMEM/F-12 อาหาร containing 3 µM CHIR99021 (a substance ที่ออกฤทธิ์ต่อ the วิถีสัญญาณ Wnt) and 25 added with lx N2 supplement, at 37°C, 5% C02• By day 27 from the start of the การเพาะเลี้ยงแบบลอย, เรเร having a characteristic polygonal cell form were observed (Fig. 3A). The expression of protein was confirmed by a cell staining method. As a result, the cells 30 were positive.to the transcription factor Mitf, which is a retinal pigment epithelial cell marker (Fig. 3B) and positive to Z0-1, which is a tight junction marker (Fig. 3C), from which it was confirmed that the obtained cells were เรเร.