Jing Wang1, Ting-Jun Fan2, Xiu-Xia Yang2, Shi-Min Chang1. 1. College of Life Sciences, Langfang Normal University, Langfang 065000, Hebei Province, China. 2. Key Laboratory for Corneal Tissue Engineering, College of Marine Life Sciences, Ocean University of China, Qingdao 266003, Shandong Province, China.
Abstract
AIM: To investigate the morphological altering effect of transforming growth factor-β2 (TGF-β2) on untransfected human corneal endothelial cells (HCECs) in vitro. METHODS: After untransfected HCECs were treated with TGF-β2 at different concentrations, the morphology, cytoskeleton distribution, and type IV collagen expression of the cells were examined with inverted contrast light microscopy, fluorescence microscopy, immunofluorescence or Western Blot. RESULTS: TGF-β2 at the concentration of 3-15 µg/L had obviously alterative effects on HCECs morphology in dose and time-dependent manner, and 9 µg/L was the peak concentration. TGF-β2 (9 µg/L) altered HCE cell morphology after treatment for 36h, increased the mean optical density (P<0.01) and the length of F-actin, reduced the mean optical density (P<0.01) of the collagen type IV in extracellular matrix (ECM) and induced the rearrangement of F-actin, microtubule in cytoplasm and collagen type IV in ECM after treatment for 72h. CONCLUTION: TGF-β2 has obviously alterative effect on the morphology of HCECs from polygonal phenotype to enlarged spindle-shaped phenotype, in dose and time-dependence manner by inducing more, elongation and alignment of F-actin, rearrangement of microtubule and larger spread area of collagen type IV.
AIM: To investigate the morphological altering effect of transforming growth factor-β2 (TGF-β2) on untransfected human corneal endothelial cells (HCECs) in vitro. METHODS: After untransfected HCECs were treated with TGF-β2 at different concentrations, the morphology, cytoskeleton distribution, and type IV collagen expression of the cells were examined with inverted contrast light microscopy, fluorescence microscopy, immunofluorescence or Western Blot. RESULTS: TGF-β2 at the concentration of 3-15 µg/L had obviously alterative effects on HCECs morphology in dose and time-dependent manner, and 9 µg/L was the peak concentration. TGF-β2 (9 µg/L) altered HCE cell morphology after treatment for 36h, increased the mean optical density (P<0.01) and the length of F-actin, reduced the mean optical density (P<0.01) of the collagen type IV in extracellular matrix (ECM) and induced the rearrangement of F-actin, microtubule in cytoplasm and collagen type IV in ECM after treatment for 72h. CONCLUTION: TGF-β2 has obviously alterative effect on the morphology of HCECs from polygonal phenotype to enlarged spindle-shaped phenotype, in dose and time-dependence manner by inducing more, elongation and alignment of F-actin, rearrangement of microtubule and larger spread area of collagen type IV.
Entities:
Keywords:
F-actin; collagen type IV; human corneal endothelial cell; microtubule; transforming growth factor-β2
Authors: Ulrike B Kottler; Anselm G M Jünemann; Thomas Aigner; Matthias Zenkel; Carmen Rummelt; Ursula Schlötzer-Schrehardt Journal: Exp Eye Res Date: 2005-01 Impact factor: 3.467