ObjectiveTo explore the culture of human eye trabecular meshwork cells in vitro, and establish a new method of identifying cells according to its characters.

MethodsThe trabecular meshwork tissue was isolated from the eyeballs of patients with ocular rupture injury, and corneal ring left after corneal transplantation, and the human primary trabecular meshwork cells were cultured in vitro using tissue block adherence and digestion methods.The cell growth was observed under an inverted microscope, and the growth rate was measured using CCK8.The fiber connection protein, type Ⅳ collagen protein, layer adhesion protein and water channel protein-1 were identified using cell immunofluorescence staining.The cultured cells were induced using 100 nml/L of dexamethasone for 10 d, and the mRNA and protein expression levels of myocilin were detected using fluorescence quantitative PCR and western blotting, respectively.

ResultsAfter culturing for 1 to 2 weeks, the cells began to grow out from the side of the trabecular meshwork, and gradually increased.After passaging, the trabecular meshwork cells grew faster on 1-4 days, and slower on 5-7 days, which still significantly higher than that on day 4(P < 0.01).The staining of fibronectin, collagen-Ⅳ, laminin and aquaporin 1 in cultured cells were positive using immunofluorescence.After dexamethasone inducing, the myocilin mRNA and protein expression levels in trabecular meshwork cells significantly increased compared with the control group(P < 0.01).

ConclusionsThe primary trabecular meshwork cells can be identified by detecting their characteristics.