Background Tear deficiency due to lacrimal gland (LG) dysfunction is one of the major causes of dry eye disease (DED). Therefore, LG stem cell-based therapies have been extensively reported to regenerate injured lacrimal tissue; however, the number of stem cells in the LG tissue is low, and 2D long-term cultivation reduces the differentiation capacity of stem cells. Nevertheless, 3D LG organoids could be an alternative for a DED therapy because it is capable of prolonged growth while maintaining the characteristics of the LG tissue. Here, we report the development of LG organoids and their application as cell therapeutics. Methods Digested cells from human LG tissue were mixed with Matrigel and cultured in five different media modified from human prostate/salivary organoid culture media. After organoid formation, the growth, specific marker expression, and histological characteristics were analyzed to authenticate the formation of LG organoids. The secretory function of LG organoids was confirmed  through calcium influx or proteomics analysis after pilocarpine treatment. To explore the curability of the developed organoids, mouse-derived LG organoids were fabricated and transplanted into the lacrimal tissue of a mouse model of DED. Results The histological features and specific marker expression of LG organoids were similar to those of normal LG tissue. In the pilocarpine-treated LG organoid, levels of internal Ca2+ ions and β-hexosaminidase, a lysosomal protein in tear fluid, were increased. In addition, the secreted proteins from pilocarpine-treated lacrimal organoids were identified through proteomics. More than 70% of the identified proteins were proven to exosome through gene ontology analysis. These results indicate that our developed organoid was pilocarpine reactive, demonstrating the function of LG. Additionally, we developed LG organoids from patients with Sjogren’s syndrome patients (SS) and confirmed that their histological features were similar to those of SS-derived LG tissue. Finally, we confirmed that the mouse LG organoids were well engrafted in the lacrimal tissue two weeks after transplantation. Conclusion This study demonstrates that the established LG organoids resemble the characteristics of normal LG tissue and may be used as a therapy for patients with DED.

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Background: Tear deficiency due to dysfunction of the lacrimal gland (LG) is one of the major causes of dry eye disease (DED). Therefore, LG stem cell-based therapies have been extensively reported to regenerate injured lacrimal tissue, but the number of stem cells in the LG tissue is pretty low, and 2D long-term cultivation results in the reduction of the differentiation capacity of stem cells. Whereas, 3D LG organoids could be an alternative for a DED therapeutic method, because it is capable of prolonged growth while maintaining the characteristics of the LG tissue. In this study, we developed LG organoids and applied them as cell therapeutics. Methods: Digested cells from human LG tissue were mixed with Matrigel and cultured in five different media modified from human prostate/salivary organoid culture media. After organoid formation, the growth, expression of specific markers and histological characters were analyzed to prove the LG organoids. And then, the secretory function of LG organoids was confirmed by calcium influx or proteomics after pilocarpine treatment. To explore the curability of developed organoids, finally, mouse-derived lacrimal gland organoids were fabricated and transplanted into mouse lacrimal tissue with DED. Results: The histological features and specific marker expression of LG organoids were similar to normal LG tissue. In the pilocarpine treated LG organoid, internal Ca2+ ions and β -hexosaminidase as known a lysosomal protein in tear fluid was increased. Also, the secreted proteins from pilocarpine treated lacrimal organoid were identified through proteomics. More than 70% of the identified proteins were proven to exosome through GO analysis. These results indicate that our developed organoid has a reactivity to pilocarpine, which shows the function of the lacrimal gland. Additionally, we developed LG organoids from Sjogren’s syndrome patients (SS) and confirmed that their histological feutures similar to that of SS-derived LG tissue. Finally, we confirmed that organoids were well engrafted in the lacrimal tissue at 2 weeks after transplantation of mouse LG orgnoid. Conclusion: This current study demonstrate that our established lacrimal gland organoids resemble characteristics of normal lacrimal gland tissue and could be used as cell therapy for patients with dry eye syndrome.

Background: Tear deficiency due to dysfunction of the lacrimal gland is one of the major causes of dry eye disease (DED). Artificial tears as conventional medicine often alleviate symptoms, however, they insufficient to prevent the progression of severe DED. Lacrimal gland stem cell-based therapies have been extensively reported to regenerate injured lacrimal tissue, but the number of stem cells in the lacrimal gland is pretty low, and 2D long-term cultivation results in the reduction of the differentiation capacity of stem cells. Therefore, 3D lacrimal gland organoids could be an alternative for a DED therapeutic method, which is capable of prolonged growth while maintaining the characteristics of the lacrimal gland tissue.Method: Digested cells from human lacrimal gland tissue were mixed with Matrigel and cultured in five different media modified from human prostate/salivary organoid culture media. After organoid formation, the growth, expression of specific markers and histological characters were analyzed to prove the lacrimal gland organoids. And then, the secretory function of lacrimal gland organoids was confirmed by calcium influx or proteomics after pilocarpine treatment.Result: Human lacrimal gland organoids were formed and cultured under a modified salivary organoid culture medium, and their histological features and gene expression pattern were similar to normal lacrimal gland tissue. After pilocarpine treatment, their secretory function was confirmed and the secretomes were analyzed by proteomics. More than 70% of the identified proteins were proven to exosome through GO analysis. Additionally, to explore the curability of developed organoids, mouse-derived lacrimal gland organoids were fabricated and transplanted into mouse lacrimal tissue with DED, and we confirmed that organoids were well engrafted in the lacrimal tissue.Conclusion: These results demonstrate that our established lacrimal gland organoids resemble characteristics of normal lacrimal gland tissue and could be used as cell therapy for patients with dry eye syndrome.