Isolation, Culture, and Characterization of Human Fetal Trabecular Meshwork Cells

Lin, Shan; Lee, On-Tat; Minasi, Petros; Wong, Joshua L. C.

doi:10.1080/02713680601107058

Cited by 19 publications

(14 citation statements)

References 13 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The morphology of the cells we used in our study was similar to that reported by Rhee et al [24], Morgan et al [19], and Lin et al [22], where they were small in size, slightly elongated, exhibited a moderate to low number of processes, and their cell bodies were both rounder and wider than those of scleral fibroblasts. The predominant marker of TM cells is increased myocilin expression following treatment with Dex [10, 25, 26].…”

Section: Resultssupporting

confidence: 82%

“…In the study, we used TM cells from a 25-year-old donor and two human fetal TM cells since TM cells from younger donors have increased and consistent doubling rates and greater passage capacity [22, 23]. The morphology of the cells we used in our study was similar to that reported by Rhee et al [24], Morgan et al [19], and Lin et al [22], where they were small in size, slightly elongated, exhibited a moderate to low number of processes, and their cell bodies were both rounder and wider than those of scleral fibroblasts.…”

Section: Resultsmentioning

confidence: 99%

See 1 more Smart Citation

Small-molecule inhibition of Wnt signaling abrogates dexamethasone-induced phenotype of primary human trabecular meshwork cells

Ahadome

Zhang

Tannous

et al. 2017

Experimental Cell Research

View full text Add to dashboard Cite

Trabecular meshwork (TM) cells are the governing regulators of the TM structure. When the functionality of these cells is impaired, the structure of the TM is perturbed, which often results in increased ocular hypertension. High intraocular pressure is the most significant risk factor for steroid-induced glaucoma. Dexamethasone (Dex) induced phenotype of TM cells is widely utilized as a model system to gain insight into mechanisms underlying damaged TM in glaucoma. In this study, to assess the possible role of the abnormal Wnt signaling in steroid-induced glaucoma, we analyzed the effects of small-molecule Wnt signaling modulators on Dex-induced expression extracellular matrix proteins of primary human TM cells. While Dex-treated TM cells exhibited increased collagen and fibronectin expression, we found that Wnt signaling inhibitor 3235-0367 suppressed these Dex-induced effects. We therefore propose that Wnt signaling plays an important role in Dex-mediated impairment of TM cell functions. Moreover, the use of small molecule Wnt signaling inhibitors to treat TM cells may provide us an opportunity of restoring TM tissue in steroid-induced glaucoma.

show abstract

Section: Resultssupporting

confidence: 82%

Section: Resultsmentioning

confidence: 99%

Small-molecule inhibition of Wnt signaling abrogates dexamethasone-induced phenotype of primary human trabecular meshwork cells

Ahadome

Zhang

Tannous

et al. 2017

Experimental Cell Research

View full text Add to dashboard Cite

show abstract

“…In fact, the increased expression of MYOC caused by Dex is currently used to characterize cultured TM cells (Lin et al, 2007; Mao et al, 2012; Stamer et al, 1998). In the present study and others, the profile of MYOC up-regulation by DEX was dose- and time-dependent, very similar to the course of development of steroid-induced glaucoma (Nguyen et al, 1998) (Shepard et al, 2001; Tamm et al, 1999); suggesting that MYOC plays a role in GC-induced glaucoma (Shepard et al, 2001).…”

Section: Discussionmentioning

confidence: 99%

Differential response and withdrawal profile of glucocorticoid-treated human trabecular meshwork cells

Cui

Dismuke

et al. 2017

Experimental Eye Research

View full text Add to dashboard Cite

The goal of the study was to examine secreted protein response and withdrawal profiles from cultured human trabecular meshwork (HTM) cells following short- and long-term glucocorticoid treatment. Primary cultures of five human HTM cell strains isolated from 5 different individual donor eyes were tested. Confluent HTM cells were differentiated in culture media containing 1% FBS for at least one week, and then treated with Dexamethasone (Dex, 100 nM) 3 times/week for 1 or 4 weeks. Cell culture supernatant was collected 3 times per week for 8 weeks. Secretion profiles of myocilin (MYOC), matrix metalloproteinase-2 (MMP2) and fibronectin (FN) were determined by Western blot analysis and MMP2 activity by zymography. Dex treatment reduced MMP2 expression and activity, returning to normal levels shortly after Dex withdrawal in 5 HTM cell strains. All five cell strains significantly upregulated MYOC in response to Dex treatment by an average of 17-fold, but recovery to basal levels after Dex withdrawal took vastly different periods of time depending on cell strain and treatment duration. Dex treatment significantly increased FN secretion in all strains but one, which decreased FN secretion in the presence of Dex. Interestingly, secretion of FN and MYOC negatively correlated during a 4 week recovery period following 4 weeks of Dex treatment. Taken together, the time course and magnitude of response and recovery for three different secreted, extracellular matrix-associated proteins varied greatly between HTM cell strains, which may underlie susceptibility to glucocorticoid-induced ocular hypertension.

show abstract

“…Aquaporins facilitate regulated water permeability in various sensory and non-sensory tissues, a process which may be crucial for the maintenance of normal tissue mechanics and the ionic balance of fluids. Aqp1 is expressed by cells in the trabecular meshwork of the eye, where it is associated with the regulation of aqueous humor volume and intra-ocular pressure (Verkman 2003;Lin et al 2007). Cells in this meshwork also express cx43, αsma, nmII, and vimentin (Kimura et al 2000;Zhang and Rao 2005;Lin et al 2007;Inoue et al 2010).…”

Section: Contractile Type III Fibrocytes Can Be Derived From the Cultmentioning

confidence: 99%

“…Aqp1 is expressed by cells in the trabecular meshwork of the eye, where it is associated with the regulation of aqueous humor volume and intra-ocular pressure (Verkman 2003;Lin et al 2007). Cells in this meshwork also express cx43, αsma, nmII, and vimentin (Kimura et al 2000;Zhang and Rao 2005;Lin et al 2007;Inoue et al 2010). These properties suggest parallels in the functions mediated by the trabecular meshwork and the continuous helical strip formed by type III fibrocytes.…”

Section: Contractile Type III Fibrocytes Can Be Derived From the Cultmentioning

confidence: 99%

Contractility in Type III Cochlear Fibrocytes Is Dependent on Non-muscle Myosin II and Intercellular Gap Junctional Coupling

Kelly

Forge

Jagger

2012

JARO

View full text Add to dashboard Cite

The cochlear spiral ligament is a connective tissue that plays diverse roles in normal hearing. Spiral ligament fibrocytes are classified into functional subtypes that are proposed to carry out specialized roles in fluid homeostasis, the mediation of inflammatory responses to trauma, and the fine tuning of cochlear mechanics. We derived a secondary sub-culture from guinea pig spiral ligament, in which the cells expressed protein markers of type III or "tension" fibrocytes, including non-muscle myosin II (nmII), α-smooth muscle actin (αsma), vimentin, connexin43 (cx43), and aquaporin-1. The cells formed extensive stress fibers containing αsma, which were also associated intimately with nmII expression, and the cells displayed the mechanically contractile phenotype predicted by earlier modeling studies. cx43 immunofluorescence was evident within intercellular plaques, and the cells were coupled via dye-permeable gap junctions. Coupling was blocked by meclofenamic acid (MFA), an inhibitor of cx43-containing channels. The contraction of collagen lattice gels mediated by the cells could be prevented reversibly by blebbistatin, an inhibitor of nmII function. MFA also reduced the gel contraction, suggesting that intercellular coupling modulates contractility. The results demonstrate that these cells can impart nmII-dependent contractile force on a collagenous substrate, and support the hypothesis that type III fibrocytes regulate tension in the spiral ligament-basilar membrane complex, thereby determining auditory sensitivity.

show abstract

Isolation, Culture, and Characterization of Human Fetal Trabecular Meshwork Cells

Cited by 19 publications

References 13 publications

Small-molecule inhibition of Wnt signaling abrogates dexamethasone-induced phenotype of primary human trabecular meshwork cells

Small-molecule inhibition of Wnt signaling abrogates dexamethasone-induced phenotype of primary human trabecular meshwork cells

Differential response and withdrawal profile of glucocorticoid-treated human trabecular meshwork cells

Contractility in Type III Cochlear Fibrocytes Is Dependent on Non-muscle Myosin II and Intercellular Gap Junctional Coupling

Contact Info

Product

Resources

About