Three-Dimensional Culture of Functional Adult Rabbit Lacrimal Gland Epithelial Cells on Decellularized Scaffold

Lin, Hui-Kuan; Sun, Guoying; He, Hong; Botsford, Benjamin; Li, Mackenzie; Elisseeff, Jennifer H.; Yiu, Samuel C.

doi:10.1089/ten.tea.2015.0286

Cited by 30 publications

(25 citation statements)

References 34 publications

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“…To date, many tissues have been successfully decellularized, including rat heart [10], rat liver [11], rat lung [12], rat kidney [13], and mouse brain [14]. In addition, various groups have carried out decellularization of tissues in the eye, including porcine cornea for corneal grafts [15], porcine conjunctiva for tissue engineering studies [16], rabbit lacrimal glands to study lacrimal gland restoration [17], bovine retina for the production of retinal progenitor cell culture films [18], and human Bruch's membrane to study RPE cell seeding [19],[20]. In contrast, to our knowledge, methods for decellularizing the choroid for the development of CEC transplantation protocols have not been described.…”

Section: Introductionmentioning

confidence: 99%

Preparation and evaluation of human choroid extracellular matrix scaffolds for the study of cell replacement strategies

et al. 2017

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Endothelial cells (ECs) of the choriocapillaris are one of the first cell types lost during age-related macular degeneration (AMD), and cell replacement therapy is currently a very promising option for patients with advanced AMD. We sought to develop a reliable method for the production of human choroidal extracellular matrix (ECM) scaffolds, which will allow for the study of choroidal EC (CEC) replacement strategies in an environment that closely resembles the native tissue. Human RPE/choroid tissue was treated sequentially with Triton X-100, SDS, and DNase to remove all native cells. While all cells were successfully removed from the tissue, collagen IV, elastin, and laminin remained, with preserved architecture of the acellular vascular tubes. The ECM scaffolds were then co-cultured with exogenous ECs to determine if the tissue can support cell growth and allow EC reintegration into the decellularized choroidal vasculature. Both monkey and human ECs took up residence in the choriocapillary tubes of the decellularized tissue. Together, these data suggest that our decellularization methods are sufficient to remove all cellular material yet gentle enough to preserve tissue structure and allow for the optimization of cell replacement strategies.

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Section: Introductionmentioning

confidence: 99%

Preparation and evaluation of human choroid extracellular matrix scaffolds for the study of cell replacement strategies

et al. 2017

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“…The resulting supernatants were analyzed for NAG catalytic activity with a NAG assay kit (Sigma-Aldrich). The absorbance at 405 nm or fluorescent intensity was measured using Synergy 2 microplate reader (BioTek, Winooski, VT) as previously described 18 . Total protein content in the samples was measured by Pierce BCA protein assay kit (Thermo Fisher Scientific).…”

Section: Methodsmentioning

confidence: 99%

“…Considering the ocular surface and tear film, researchers have been investigating the in vitro modeling of several components individually, including conjunctival epithelium 17 and lacrimal glands 18, 19 . In these models, cell morphology and phenotype resembled the equivalent cells in vivo .…”

Section: Introductionmentioning

confidence: 99%

An In Vitro Model for the Ocular Surface and Tear Film System

Yin

Grant

et al. 2017

Sci Rep

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Dry eye is a complicated ocular surface disease whose exact pathogenesis is not yet fully understood. For the therapeutic evaluation and pathogenesis study of dry eye, we established an in vitro three-dimensional (3D) coculture model for the ocular surface. It is composed of rabbit conjunctival epithelium and lacrimal gland cell spheroids, and recapitulates the aqueous and mucin layers of the tear film. We first investigated the culture conditions for both cell types to optimize their secretory functions, by employing goblet cell enrichment, air-lifting culture, and 3D spheroid formation techniques. The coculture of the two cell components leads to elevated secretion and higher expression of tear secretory markers. We also compared several coculture systems, and found that direct cell contact between the two cell types significantly increased tear secretion. Inflammation was induced to mimic dry eye disease in the coculture model system. Our results showed that the coculture system provides a more physiologically relevant therapeutic response compared to monocultures. Our work provides a complex 3D model as a recapitulation of the ocular surface and tear film system, which can be further developed as a model for dry eye disease and therapeutic evaluation.

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“…The same group recently described a non-immunogenic decellularized tissue matrix derived from pig lacrimal gland, which allows lacrimal gland epithelial cells seeded on this scaffold to maintain acinar-like structures and secretory ability for up to 30 days (Spaniol et al, 2015). In a similar effort, rabbit lacrimal gland progenitor cells derived from a sphere-forming culture have been shown to differentiate and display secretory function in either a decellularized lacrimal gland matrix or 3-D collagen gel (Lin et al, 2016). There are still challenges regarding optimization of the decellularization technique, the seeding process, the long-term culturing conditions, and functional validation.…”

Section: The Molecular Mechanism Of Lacrimal Gland Developmentmentioning

confidence: 99%

Lacrimal gland development: From signaling interactions to regenerative medicine

Garg

Zhang

2017

Developmental Dynamics

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The lacrimal gland plays a pivotal role in keeping the ocular surface lubricated, and protecting it from environmental exposure and insult. Dysfunction of the lacrimal gland results in deficiency of the aqueous component of the tear film, which can cause dryness of the ocular surface, also known as the aqueous-deficient dry eye disease. Left untreated, this disease can lead to significant morbidity, including frequent eye infections, corneal ulcerations and vision loss. Current therapies do not treat the underlying deficiency of the lacrimal gland, but merely provide symptomatic relief. To develop more sustainable and physiological therapies, such as in vivo lacrimal gland regeneration or bioengineered lacrimal gland implants, a thorough understanding of lacrimal gland development at the molecular level is of paramount importance. Based on the structural and functional similarities between rodent and human eye development, extensive studies have been undertaken to investigate the signaling and transcriptional mechanisms of lacrimal gland development using mouse as a model system. In this review, we describe the current understanding of the extrinsic signaling interactions and the intrinsic transcriptional network governing lacrimal gland morphogenesis, as well as recent advances in the field of regenerative medicine aimed at treating dry eye disease.

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Three-Dimensional Culture of Functional Adult Rabbit Lacrimal Gland Epithelial Cells on Decellularized Scaffold

Cited by 30 publications

References 34 publications

Preparation and evaluation of human choroid extracellular matrix scaffolds for the study of cell replacement strategies

Preparation and evaluation of human choroid extracellular matrix scaffolds for the study of cell replacement strategies

An In Vitro Model for the Ocular Surface and Tear Film System

Lacrimal gland development: From signaling interactions to regenerative medicine

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