scholarly journals Establishment of functional epithelial organoids from human lacrimal glands

2021 ◽  
Author(s):  
Sang Yun Jeong ◽  
Woo Hee Choi ◽  
Seong Gyeong Jeon ◽  
Sookon Lee ◽  
Jong-Moon Park ◽  
...  
Keyword(s):  
Stem Cells ◽  
Dry Eye ◽  
Lacrimal Gland ◽  
Culture Media ◽  
Calcium Influx ◽  
Eye Disease ◽  
Specific Marker ◽  
Gland Tissue ◽  
Lysosomal Protein

Abstract 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.

scholarly journals Establishment of Functional Epithelial Organoid From Human Lacrimal Glands

2020 ◽  
Author(s):  
Sang Yun Jeong ◽  
Woo Hee Choi ◽  
Sookon Lee ◽  
Jong-Moon Park ◽  
Mira Park ◽  
...  
Keyword(s):  
Stem Cells ◽  
Dry Eye ◽  
Lacrimal Gland ◽  
Culture Media ◽  
Conventional Medicine ◽  
Calcium Influx ◽  
Gene Expression Pattern ◽  
Secretory Function ◽  
Organoid Culture ◽  
Gland Tissue

Abstract 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.

scholarly journals Establishment of functional epithelial organoids from human lacrimal glands

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Sang Yun Jeong ◽  
Woo Hee Choi ◽  
Seong Gyeong Jeon ◽  
Sookon Lee ◽  
Jong-Moon Park ◽  
...  
Keyword(s):  
Stem Cells ◽  
Culture Media ◽  
Calcium Influx ◽  
Specific Marker ◽  
Histological Features ◽  
Hexosaminidase A ◽  
Histological Characteristics ◽  
Marker Expression ◽  
Lysosomal Protein

AbstractBackgroundTear 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.MethodsDigested 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.ResultsThe 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.ConclusionThis 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.

scholarly journals Long-term Natural History of Dry Eye Disease from the Patient's Perspective

Ophthalmology ◽  
2016 ◽  
Vol 123 (2) ◽  
pp. 425-433 ◽  
Author(s):  
Jeffrey P. Lienert ◽  
Laura Tarko ◽  
Miki Uchino ◽  
William G. Christen ◽  
Debra A. Schaumberg
Keyword(s):  
Natural History ◽  
Dry Eye ◽  
Dry Eye Disease ◽  
Eye Disease ◽  
Patient’S Perspective ◽  
History Of

scholarly journals Long-term outcome after topical ciclosporin in severe dry eye disease with a 10-year follow-up

2016 ◽  
Vol 100 (11) ◽  
pp. 1547-1550 ◽  
Author(s):  
Morgane Straub ◽  
Alain M Bron ◽  
Aurore Muselier-Mathieu ◽  
Catherine Creuzot-Garcher
Keyword(s):  
Dry Eye ◽  
Dry Eye Disease ◽  
Eye Disease ◽  
Term Outcome ◽  
Long Term Outcome

scholarly journals Lacrimal Gland Myoepithelial Cells Are Altered in a Mouse Model of Dry Eye Disease

2020 ◽  
Vol 190 (10) ◽  
pp. 2067-2079 ◽  
Author(s):  
Laura García-Posadas ◽  
Robin R. Hodges ◽  
Tor P. Utheim ◽  
Ole Kristoffer Olstad ◽  
Vanessa Delcroix ◽  
...  
Keyword(s):  
Mouse Model ◽  
Dry Eye ◽  
Lacrimal Gland ◽  
Myoepithelial Cells ◽  
Dry Eye Disease ◽  
Eye Disease

scholarly journals HIF1α-mediated TRAIL Expression Regulates Lacrimal Gland Inflammation in Dry Eye Disease

2020 ◽  
Vol 61 (1) ◽  
pp. 3
Author(s):  
Yong Woo Ji ◽  
Joon H. Lee ◽  
Eun Young Choi ◽  
Hyun Goo Kang ◽  
Kyoung Yul Seo ◽  
...  
Keyword(s):  
Dry Eye ◽  
Lacrimal Gland ◽  
Dry Eye Disease ◽  
Eye Disease ◽  
Trail Expression

scholarly journals The Role of Medications in Causing Dry Eye

2012 ◽  
Vol 2012 ◽  
pp. 1-8 ◽  
Author(s):  
Frederick T. Fraunfelder ◽  
James J. Sciubba ◽  
William D. Mathers
Keyword(s):  
Dry Eye ◽  
Ocular Surface ◽  
Dry Mouth ◽  
Eye Disease ◽  
Therapeutic Action ◽  
Common Cause ◽  
Complex Interactions ◽  
Ocular Drugs

The purpose of this paper is to review the possible role of polypharmacy in causing dry eye disease (DED), reflecting the complex interactions and complications associated with the use of multiple systemic and topical ocular medications. The pharmacological, physiological, anatomical, and histological mechanisms causing dry mouth differ little from those causing dry eye. Oral polypharmacy is the most common cause of dry mouth, but has not been investigated as a cause of dry eye. Topical ocular polypharmacy has been shown to cause DED. Information on drugs that likely cause or aggravate DED and the controversial role of preservatives in topical ocular medications are examined. Systemic or topical ocular medications and preservatives used in topical ocular drugs may cause dry eye through the drug's therapeutic action, ocular surface effects, or preservatives, and the effects probably are additive. Long-term use of topical ocular medications, especially those containing preservatives such as BAK, may play an important role in DED and the role of polypharmacy needs further study. We review possible ways to decrease the risk of medication-related dry eye.

scholarly journals Establishment of long-term serum-free culture for lacrimal gland stem cells aiming at lacrimal gland repair

2020 ◽  
Vol 11 (1) ◽  
Author(s):  
Sa Xiao ◽  
Yan Zhang
Keyword(s):  
Stem Cells ◽  
Lacrimal Gland ◽  
Progenitor Cell ◽  
Adult Mouse ◽  
3D Culture ◽  
Phenol Red ◽  
Serum Free ◽  
Ductal Cells ◽  
Cell Based Therapy

Abstract Background Aqueous-deficient dry eye disease (ADDED) resulting from dysfunction of the lacrimal gland (LG) is currently incurable. Although LG stem/progenitor cell-based therapy is considered to be a promising strategy for ADDED patients, the lack of a reliable serum-free culture method to obtain enough lacrimal gland stem cells (LGSCs) and the basic standard of LGSC transplantation are obstacles for further research. Methods Adult mouse LGSCs were cultured in Matrigel-based 3D culture under serum-free culture condition, which contained EGF, FGF10, Wnt3A, and Y-27632. LGSCs were continuously passaged over 40 times every 7 days, and the morphology and cell numbers were recorded. LGSCs were induced to differentiate to ductal cells by reducing Matrigel rigidity, while fetal bovine serum was used for the induction of acinar cells. RT-PCR or qRT-PCR analysis, RNA-sequence analysis, H&E staining, and immunofluorescence were used for characterization and examining the differentiation of LGSCs. LGSCs were allotransplanted into diseased LGs to examine the ability of repairing the damage. The condition of eye orbits was recorded using a camera, the tear production was measured using phenol red-impregnated cotton threads, and the engraftments of LGSCs were examined by immunohistochemistry. Results We established an efficient 3D serum-free culture for adult mouse LGSCs, in which LGSCs could be continuously passaged for long-term expansion. LGSCs cultured from both the healthy and ADDED mouse LGs expressed stem/progenitor cell markers Krt14, Krt5, P63, and nestin, had the potential to differentiate into acinar or ductal-like cells in vitro and could engraft into diseased LGs and relieve symptoms of ADDED after orthotopic injection of LGSCs. Conclusion We successfully established an efficient serum-free culture for adult mouse LGSCs aiming at LG repair for the first time. Our approach provides an excellent theoretical and technical reference for future clinical research for ADDED stem cell therapy.

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