人胚胎干细胞(hESC)和人诱导多能干细胞(hiPSC)衍生的视网膜色素上皮(RPE)疗法是治疗由RPE变性引起的视网膜变性疾病的有希望的替代品。从人类成人供体产生自体RPE细胞,具有避免免疫排斥反应和畸胎瘤形成的优势,是一种替代细胞资源,可获得对RPE退行性疾病的机械洞察力和测试潜在疗法。这里,我们发现,来自hESCs和成人原代角膜缘的角膜缘干细胞(LSCs)具有产生RPE细胞和角膜基质干细胞(CSSCs)的潜能.我们显示hESC-LSC衍生的RPE细胞(LSC-RPE)表达RPE标记,有吞噬功能,和综合热带因素。此外,在从LSCs分化为RPE细胞的过程中,细胞变得有色素,伴随着LSC标志物KRT15水平的降低和RPE标志物MITF水平的升高。Wnt信号通路在LSC-RPE命运转变中起作用,促进细胞核中的MITF表达,并鼓励RPE命运转变。此外,我们还表明,来自成人角膜缘的原代LSCs(pLSCs)类似于hESC-LSC可以产生RPE细胞,这得到了LSC和RPE细胞标志物(KRT15/OTX2,KRT15/MITF)共表达的支持,提示从pLSC到RPE细胞的过渡,和典型的多边形形态,黑化,RPE细胞标记基因表达(TYR,RPE65),通过ZO-1表达形成紧密连接,和最关键的吞噬功能。另一方面,hESC-LSCs和pLSCs也分化成表达干细胞标志物(PAX6,NESTIN)的CSSCs(LSC-CSSCs),介绍MSC功能,包括表面标记表达和三系分化能力,就像人类CSSC中的那些。此外,pLSC-CSSC分化成表达角膜细胞标记基因的细胞的能力(ALDH3A1,PTGDS,PDK4)表明诱导角膜细胞的潜力。这些结果表明,成体pLSC是一种替代细胞资源,其应用为预防RPE功能障碍相关的视网膜变性疾病和角膜瘢痕形成提供了新的潜在治疗途径。
Human embryonic stem cell (hESC)- and human induced pluripotent stem cell (hiPSC)-derived retinal pigment epithelium (RPE) therapies are promising alternatives for the treatment of retinal degenerative diseases caused by RPE degeneration. The generation of autologous RPE cells from human adult donors, which has the advantage of avoiding immune rejection and teratoma formation, is an alternative cell resource to gain mechanistic insight into and test potential therapies for RPE degenerative diseases. Here, we found that limbal stem cells (LSCs) from hESCs and adult primary human limbus have the potential to produce RPE cells and corneal stromal stem cells (CSSCs). We showed that hESC-LSC-derived RPE cells (LSC-RPE) expressed RPE markers, had a phagocytic function, and synthesized tropical factors. Furthermore, during differentiation from LSCs to RPE cells, cells became pigmented, accompanied by a decrease in the level of LSC marker KRT15 and an increase in the level of RPE marker MITF. The Wnt signaling pathway plays a role in LSC-RPE fate transition, promotes MITF expression in the nucleus, and encourages RPE fate transition. In addition, we also showed that primary LSCs (pLSCs) from adult human limbus similar to hESC-LSC could generate RPE cells, which was supported by the co-expression of LSC and RPE cell markers (KRT15/OTX2, KRT15/MITF), suggesting the transition from pLSC to RPE cells, and typical polygonal morphology, melanization, RPE cell marker genes expression (TYR, RPE65), tight junction formation by ZO-1 expression, and the most crucial phagocytotic function. On the other hand, both hESC-LSCs and pLSCs also differentiated into CSSCs (LSC-CSSCs) that expressed stem cell markers (PAX6, NESTIN), presented MSC features, including surface marker expression and trilineage differentiation capability, like those in human CSSCs. Furthermore, the capability of pLSC-CSSC to differentiate into cells expressing keratocyte marker genes (ALDH3A1, PTGDS, PDK4) indicated the potential to induce keratocytes. These results suggest that the adult pLSC is an alternative cell resource, and its application provides a novel potential therapeutic avenue for preventing RPE dysfunction-related retinal degenerative diseases and corneal scarring.