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.

干眼病是老龄化眼睛中最普遍的病理状况。一种潜在的治疗策略是将多能干细胞（例如人胚胎干细胞）体外产生的泪腺移植到患者体内。前述要求之一是将人类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，本研究旨在确定一组转录因子，这些转录因子的过表达可以促进人胚胎干细胞向泪腺上皮样细胞的分化。我们进行了从小鼠胚胎和成年获得的泪腺和泪腺相关器官的微阵列分析，并确定了富含泪腺上皮细胞的转录因子。然后，我们将编码这些转录因子的人类直向同源物的合成信使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确定了富含泪腺上皮细胞的16个转录因子。我们专注于三种转录因子，因为它们在其他腺体（如唾液腺）中表达，并且还已知与泪腺的发育有关。我们测试了三种转录因子和 因为它们在其他腺体（如唾液腺）中表达，并且还已知与泪腺的发育有关。我们测试了三种转录因子和 因为它们在其他腺体（如唾液腺）中表达，并且还已知与泪腺的发育有关。我们测试了三种转录因子和PAX6是人类胚胎干细胞中泪腺发育必不可少的基因。结合使用PAX6，SIX1和FOXC1会导致形态上的显着变化，即上皮标记物（如细胞角蛋白15），分支形态发生标记物（如BARX2）的延长的细胞形状和增加的表达（RNA和蛋白质）以及泪腺标志物，例如水通道蛋白5和乳铁蛋白。我们确定了一组富含泪腺上皮细胞的转录因子，并证明了这些转录因子的同时过表达可以将人类胚胎干细胞分化为泪腺上皮样细胞。这项研究表明从人类多能干细胞中产生泪腺的可能性。