Dry eye disease is the most prevalent pathological condition in aging eyes. One potential therapeutic strategy is the transplantation of lacrimal glands, generated in vitro from pluripotent stem cells such as human embryonic stem cells, into patients. One of the preceding requirements is a method to differentiate human embryonic stem cells into lacrimal gland epithelium cells. As the first step for this approach, this study aims to identify a set of transcription factors whose overexpression can promote the differentiation of human embryonic stem cells into lacrimal gland epithelium-like cells. We performed microarray analyses of lacrimal glands and lacrimal glands-related organs obtained from mouse embryos and adults, and identified transcription factors enriched in lacrimal gland epithelium cells. We then transfected synthetic messenger RNAs encoding human orthologues of these transcription factors into human embryonic stem cells and examined whether the human embryonic stem cells differentiate into lacrimal gland epithelium-like cells by assessing cell morphology and marker gene expression. The microarray analysis of lacrimal glands tissues identified 16 transcription factors that were enriched in lacrimal gland epithelium cells. We focused on three of the transcription factors, because they are expressed in other glands such as salivary glands and are also known to be involved in the development of lacrimal glands. We tested the overexpression of various combinations of the three transcription factors and PAX6, which is an indispensable gene for lacrimal glands development, in human embryonic stem cells. Combining PAX6, SIX1, and FOXC1 caused significant changes in morphology, i.e., elongated cell shape and increased expression (both RNAs and proteins) of epithelial markers such as cytokeratin15, branching morphogenesis markers such as BARX2, and lacrimal glands markers such as aquaporin5 and lactoferrin. We identified a set of transcription factors enriched in lacrimal gland epithelium cells and demonstrated that the simultaneous overexpression of these transcription factors can differentiate human embryonic stem cells into lacrimal gland epithelium-like cells. This study suggests the possibility of lacrimal glands regeneration from human pluripotent stem cells.

干眼病是老化眼睛中最常见的病理状况。一种潜在的治疗策略是将由多能干细胞（例如人类胚胎干细胞）在体外产生的泪腺移植到患者体内。前述要求之一是将人胚胎干细胞分化为泪腺上皮细胞的方法。作为该方法的第一步，本研究旨在鉴定一组转录因子，其过度表达可以促进人胚胎干细胞分化为泪腺上皮样细胞。我们对从小鼠胚胎和成体获得的泪腺和泪腺相关器官进行了微阵列分析，并鉴定了泪腺上皮细胞中富含的转录因子。然后，我们将编码这些转录因子的人类直系同源物的合成信使RNA转染到人类胚胎干细胞中，并通过评估细胞形态和标记基因表达来检查人类胚胎干细胞是否分化为泪腺上皮样细胞。泪腺组织的微阵列分析鉴定出泪腺上皮细胞中富集的 16 种转录因子。我们重点关注其中三个转录因子，因为它们在唾液腺等其他腺体中表达，并且也已知参与泪腺的发育。我们测试了三种转录因子和PAX6 （泪腺发育不可或缺的基因）的各种组合在人胚胎干细胞中的过度表达。 PAX6 、 SIX1和FOXC1的组合引起形态的显着变化，即，细长的细胞形状和上皮标记物（如细胞角蛋白15）、分支形态发生标记物（如BARX2 ）以及泪腺标记物（如水通道蛋白5和乳铁蛋白）的表达增加（RNA和蛋白质）。我们鉴定了一组在泪腺上皮细胞中富集的转录因子，并证明这些转录因子的同时过表达可以将人胚胎干细胞分化为泪腺上皮样细胞。这项研究表明人类多能干细胞有可能再生泪腺。