Establishing and characterizing lacrispheres from human lacrimal gland for potential clinical application

Tiwari, Shubha; Nair, Rohini; Vamadevan, Praseeda; Ali, Mohammad Javed; Naik, Milind N.; Honavar, Santosh G; Vemuganti, Geeta K.

doi:10.1007/s00417-018-3926-8

Cited by 17 publications

(19 citation statements)

References 18 publications

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“…As prerequisites for cell therapy, the plausibility of using lacrimal organoids containing functional stem cells to rescue and repair to obtain functionally competent cells should be studied. Such studies could contribute to regeneration of damaged tissues [23].…”

Section: Discussionmentioning

confidence: 99%

“…Contrarily to lacrimal gland cultures growing a monolayer in order of preference on Matrigel, collagen, and HAM in two to three weeks, the formation of spheroids including a mixed population of stem cells and differentiated cells has been reported in salivary gland cultures [22] and prostate spheres [23]. A single study has been conducted on spheroidal aggregation of rabbit lacrimal gland cells grown in a microgravity environment of a rotary cell culture system [24].…”

Section: Introductionmentioning

confidence: 99%

“…Under certain culture conditions, suspension 3D cultures of human "lacrispheres" could be maintained and propagated for 3-4 weeks. In addition, lacrimal spheres could secrete quanti able levels of tear protein into conditioned media [23]. Lacrimal gland epithelial cells can make 'spherules' with channel like connections.…”

Section: Introductionmentioning

confidence: 99%

“…Regarding structural and functional loss of the lacrimal gland, one promising option for managing this condition could be replacement of the gland and restoration of its function through cell therapy [23].…”

Section: Introductionmentioning

confidence: 99%

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Establishment of Functional Epithelial Organoid From Human Lacrimal Glands

Jeong

Choi

Lee

et al. 2020

Preprint

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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.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Establishment of Functional Epithelial Organoid From Human Lacrimal Glands

Jeong

Choi

Lee

et al. 2020

Preprint

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“…Many studies utilize DMEM for the culture of thyroid cell lines [17]. While HDM was originally designed for the culture of hepatocytes, it has also been reported to be suitable for the culture of epithelial cells and certain kinds of gland cells, such as hepatic epithelial cells [15], salivary epithelial cells [18] and primary human lacrimal gland cells [19]. Similarly, thyroid cells, which are follicle epithelial cells isolated from the thyroid gland, may also be well-cultured and easily proliferate in epithelial cell culture medium.…”

Section: Discussionmentioning

confidence: 99%

Fabrication of Functional Cell Sheets with Human Thyrocytes from Non-Tumorous Thyroid Tissue

Huang

Yamanouchi

Sakai

et al. 2019

Tissue Eng Regen Med

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BACKGROUND: Engineered cell sheet transplantation has been considered an alternative physiological therapy for endocrine disorders. In this study, we attempted to fabricate functional human thyroid cell sheets using the engineering technology by culturing primary thyrocytes in free-feeder monolayers and assessed their proliferation and function in two different media. METHODS: The non-tumorous tissues (approximately 2 g) were dissected during surgery. Primary human thyroid cells were isolated by mechanical dispersion and treatment with isolation solution. The cells were cultured on tissue culture dishes or temperature-responsive culture dishes to induce the formation of detached cell sheets. RESULTS: Primary thyroid cells isolated from nine patients were positive for thyroid transcription factor 1, thyroglobulin (TG) and cytokeratin 7. Cell sheets with follicles were fabricated by cells incubated in both Dulbecco's Modified Eagle Medium (DMEM) and hepatocyte-defined medium (HDM) culture medium. The diameter and thickness of sheets fabricated in HDM were larger and thicker than those fabricated from DMEM. Furthermore, the cells incubated in HDM secreted higher levels of fT3 and fT4 than those incubated in DMEM. The thyroid peroxidase and TG mRNA of cells maintained in HDM were higher than those in cells maintained in DMEM. CONCLUSION: HDM appears suitable as a culture medium for maintaining primary thyrocytes and fabricating functional cell sheets. These in vitro findings may contribute to the development of appropriate culture conditions for human thyrocytes as well as engineered functional cell sheets.

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