Serum-Induced Differentiation of Human Meibomian Gland Epithelial Cells

Sullivan, David A.; Yang, Lijun; Kam, Wendy R.; Ding, Juan; Green, Karin M.; Shaffer, Scott A.; Hatton, Mark P.; Liu, Shaohui

doi:10.1167/iovs.13-13407

Cited by 52 publications

(56 citation statements)

References 55 publications

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“…Such reversibility has also been observed in primary human corneal cells 30, indicating that the immortalized HMGECs share characteristics of primary cells. Indeed, we have previously found that many of the genetic responses of immortalized HMGECs are the same as those of primary HMGECs 18. These findings suggest that this MG cell line, which features a normal karyotype, represents an appropriate model in vitro to study HMGEC dynamics 16.…”

Section: Discussionmentioning

confidence: 67%

“…After reaching 60%–70% confluence, cells were placed for varying intervals in one of the following media: KSFM alone, PM, or serum‐containing medium composed of 10% fetal bovine serum (Thermo Fisher Scientific) in equal volumes of Dulbecco's modified Eagle's medium and Ham's F12 (differentiation medium [DM], Mediatech, Inc., a Corning Subsidiary, Manassas, VA) to induce cell differentiation 18. To accelerate differentiation of HMGECs, 10 μg/mL azithromycin (AZM; Santa Cruz Biotechnology, Dallas, TX) was added to the DM 19.…”

Section: Methodsmentioning

confidence: 99%

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Biomarkers for Progenitor and Differentiated Epithelial Cells in the Human Meibomian Gland

Xie

Sullivan

Chen

et al. 2018

Stem Cells Translational Medicine

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The meibomian gland (MG) is a sebaceous gland that secretes through a holocrine process. Because such secretion requires the destruction of MG acinar epithelial cells, they need constant renewal and differentiation. The processes that promote these regenerative events in the human MG are unknown, nor is it known how to distinguish MG progenitor and differentiated cells. We discovered that Lrig1 and DNase2 serve as biomarkers for human MG progenitor and differentiated cells, respectively. Lrig1 is expressed in MG basal epithelial cells in the acinar periphery, a location where progenitor cells originate in sebaceous glands. DNase2 is expressed in the differentiated epithelial cells of the MG central acinus. Furthermore, proliferation stimulates, and differentiation suppresses, Lrig1 expression in human MG epithelial cells. The opposite is true for DNase2 expression. Our biomarker identification may have significant value in clinical efforts to restore MG function and to regenerate MGs after disease‐induced dropout. Stem Cells Translational Medicine 2018;7:887–892

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Section: Discussionmentioning

confidence: 67%

Section: Methodsmentioning

confidence: 99%

Biomarkers for Progenitor and Differentiated Epithelial Cells in the Human Meibomian Gland

Xie

Sullivan

Chen

et al. 2018

Stem Cells Translational Medicine

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“…However, there have been no reports so far that would demonstrate that MG-derived cell cultures in vitro were able to produce lipids that would be identical (or even similar) to the mature MG secretions described in this paper. Indeed, the reported lipidomes of the cell cultures were much closer to the regular lipid profiles of non-differentiated epithelial cells that are enriched with PL and TAG, but lack WE, CE and other typical components of meibum [74–76]. Thus, it remains unknown what regulatory factors are responsible for triggering meibogenesis, which our current mechanistic approach (Fig.…”

Section: Discussionmentioning

confidence: 72%

Dissecting lipid metabolism in meibomian glands of humans and mice: An integrative study reveals a network of metabolic reactions not duplicated in other tissues

Butovich

McMahon

Wojtowicz

et al. 2016

Biochimica et Biophysica Acta (BBA) - Molecular and Cell Biolog

118

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Lipids comprise the bulk of the Meibomian gland secretion (meibum) which is produced by meibocytes. Complex arrays of lipogenic reactions in Meibomian glands, which we collectively call meibogenesis, have not been explored on molecular level yet. Our goals were to elucidate the possible biosynthetic pathways that underlie generation of meibum, reveal similarities in, and differences between, lipid metabolism in Meibomian glands and other organs and tissues, and integrate Meibomian gland studies into the field of general metabolomics. Specifically, we have conducted detailed analyses of human and mouse specimens using genomic, immunohistochemical, and lipidomic approaches. Among equally highly expressed genes found in Meibomian glands of both species were those related to fatty acid elongation, branching, desaturation, esterification, reduction of fatty acids to alcohols, and cholesterol biosynthesis. Importantly, corresponding lipid products were detected in meibum of both species using lipidomic approaches. For the first time, a cohesive, unifying biosynthetic scheme that connects genomic, lipidomic, and immunohistochemical observations is outlined and discussed.

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“…14 Previous studies, including some from our group, showed the functional influence of cultivation conditions, 15,16 neurotransmitters, 17,18 growth factors, 19,20 x-3 fatty acids, 21 sex hormones, 22,23 and pharmacologic substances 24 on HMGEC. However, all of these studies rely on 2D cell culture conditions or limited animal experiments.…”

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confidence: 94%

Differentiation Patterns of Immortalized Human Meibomian Gland Epithelial Cells in Three-Dimensional Culture

Asano

Hampel

Garreis

et al. 2018

Invest. Ophthalmol. Vis. Sci.

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PURPOSE. To establish a simplified three-dimensional (3D) meibomian gland culture model using a meibomian gland epithelial cell (HMGEC) line that might be a useful tool to gain deeper insights into meibomian gland dysfunction. For this purpose, 3D differentiation patterns and growth characteristics of HMGECs were studied on various membranes/scaffolds as well as in hanging drops.METHODS. Several types of inserts consisting of different materials (Millicell-HA, Millicell-PCF, ThinCert, and Alvetex) as well as hanging drop culture were analyzed. Culture conditions were optimized employing exposure to air (air-lift) and different cell culture media for a maximum of 28 days. To characterize cell differentiation in the developed 3D model, the expression pattern of cytokeratins was investigated by immunohistochemistry. Sudan III staining was performed for detection of lipid formation and transmission electron microscopy (TEM) was used for ultrastructural analysis. RESULTS.Only Alvetex scaffolds and the hanging drop method revealed satisfactory results with regard to 3D culture. Continuous use of proliferation medium (serum-free keratinocyte medium containing epidermal growth factor and bovine pituitary extract) and air-lift were important steps for HMGEC differentiation in 3D culture. However, HMGECs only reached a differentiating state and never became mature or hypermature. When cultured in hanging drops, HMGECs showed serum-induced keratinization processes. CONCLUSIONS.HMGECs have the capability to differentiate in a long-term 3D culture, especially when adapted to an air-rich environment. However, even in the 3D format, HMGECs only reach a state of differentiating meibocytes.

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Serum-Induced Differentiation of Human Meibomian Gland Epithelial Cells

Abstract: Overall, our findings indicate that immortalized human meibomian gland epithelial cells may serve as an ideal preclinical model to identify factors that control cellular differentiation in the meibomian gland.

Cited by 52 publications

References 55 publications

Biomarkers for Progenitor and Differentiated Epithelial Cells in the Human Meibomian Gland

Biomarkers for Progenitor and Differentiated Epithelial Cells in the Human Meibomian Gland

Dissecting lipid metabolism in meibomian glands of humans and mice: An integrative study reveals a network of metabolic reactions not duplicated in other tissues

Differentiation Patterns of Immortalized Human Meibomian Gland Epithelial Cells in Three-Dimensional Culture

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