Department of Cellular Biotechnology, Cell Science Research Center, Royan Institute for Biotechnology, ACECR, Isfahan, Iran; Department of Developmental Biology, University of Science and Culture, Tehran, Iran.
Department of Cellular Biotechnology, Cell Science Research Center, Royan Institute for Biotechnology, ACECR, Isfahan, Iran.
Differentiation. 2018 May-Jun;101:8-15. doi: 10.1016/j.diff.2018.02.003. Epub 2018 Mar 2.
Recently, we have found that human stem cells from apical papilla (SCAP) show a stromal cell-derived inducing activity (SDIA). To examine SDIA competence for retinal cells differentiation, we co-cultured SCAP with human pluripotent stem cells (hPSCs). In comparison with Matrigel-cultured hPSCs, SCAP significantly induces hPSCs to differentiate into rostral neural cells as demonstrated by upregulation of OTX2 and PAX6 and down-regulation of EN1, HOXB4 and HOXC8. Furthermore, the differentiated cells on SCAP significantly expressed eye-field markers, RAX, PAX6, LHX2 and SIX3 and showed five folds pigmented colonies. The generated hPSC-retinal pigmented epithelium (RPE) was hexagonal and highly expressed related markers, ZO-1, RPE65, BEST, CRALBP and MITF. They were able to phagocytose latex beads. Moreover, the assessment of the isolated neural tube-like structures on SCAP showed the expression of retinal progenitor cells (RPCs) - SIX3, RAX, and PAX6. SCAP highly expressed DKK3 and SFRP2, Wnt inhibitor factors and their target genes, Cyclin D1 and c-Myc were down-regulated significantly on SCAP. These results showed SCAP promoted the differentiation of hPSCs into retinal cells (RPE and RPCs) possibly through inhibition of Wnt signaling pathway. This simple and efficient approach provides human RPE generation for developing therapies for diseases such as age-related macular degeneration.
最近，我们发现人根尖乳头干细胞（SCAP）表现出基质细胞衍生诱导活性（SDIA）。为了研究 SDIA 对视网膜细胞分化的能力，我们将 SCAP 与人多能干细胞（hPSC）共培养。与 Matrigel 培养的 hPSC 相比，SCAP 显著诱导 hPSC 分化为头侧神经细胞，表现为 OTX2 和 PAX6 的上调和 EN1、HOXB4 和 HOXC8 的下调。此外，在 SCAP 上分化的细胞显著表达眼区标记物 RAX、PAX6、LHX2 和 SIX3，并表现出五倍的色素斑集落。生成的 hPSC 视网膜色素上皮（RPE）呈六方形状，高度表达相关标记物 ZO-1、RPE65、BEST、CRALBP 和 MITF。它们能够吞噬乳胶珠。此外，对 SCAP 上分离的神经管样结构的评估表明，视网膜祖细胞（RPCs）-SIX3、RAX 和 PAX6 的表达。SCAP 高度表达 Wnt 抑制因子 DKK3 和 SFRP2，其靶基因 Cyclin D1 和 c-Myc 在 SCAP 上显著下调。这些结果表明，SCAP 可能通过抑制 Wnt 信号通路促进 hPSC 分化为视网膜细胞（RPE 和 RPCs）。这种简单有效的方法为开发治疗年龄相关性黄斑变性等疾病的治疗方法提供了人 RPE 的生成。
