Extended Data Fig. 1: Differentiation of multiple ocular cell lineages in the SEAM.
From: Generation of 3D lacrimal gland organoids from human pluripotent stem cells
a, Schematic illustration of the method for inducing lacrimal-gland-like clusters in a SEAM. b, Gene expression of three germ layer markers in 2- and 4-week differentiated SEAMs (n = 4 biological replicates). Data are presented as mean ± s.d. c–d, Immunostaining for PAX6 (green) and AQP5 (red) in a 10-week differentiated SEAM. Magnified views of the dashed areas in (c) are shown in (d). n = 3 distinct samples. Nuclei, blue. Scale bars, 100 μm. e, Immunostaining for CD44 (green) and HTN1 (red) in a thin section of lacrimal-gland-like clusters in a 14-week differentiated SEAM. n = 3 biological replicates. BF, bright field; Scale bar, 100 μm. f, Flow cytometric analysis of SSEA-4, ITGB4, and CD200 in isolated lacrimal-gland-like clusters induced in SEAMs after 11 weeks of culture. Four populations in the CD200– cells are categorized as P1–P4. n = 2 biological replicates. Scale bar, 200 μm. OESCs, ocular surface epithelial stem cells. g, 3D culture of the sorted cell populations (P1–P4) to induce lacrimal-gland-like organoids at day 3 (n = 3 biological replicates). Scale bars, 1,000 μm. h, Screening for organoid induction medium using a combination of lacrimal gland-related cytokines and Y-27632. n = 2 biological replicates. Scale bars, 1,000 μm. i, Comparison of lacrimal gland-related gene expression in the organoids cultured in medium containing either Y-27632 and EGF (YE), or Y-27632, EGF, KGF, BMP7 and FGF10 (YEKBF). *p < 0.05 (n = 4 biological replicates; NS, not significant). Data are presented as mean ± s.d. Exact p values are provided in the Source Data.
