[Calf trabecular cell culture in vitro, morphological and cytoskeletal ultrastructures and kinetics investigation].
OBJECTIVE
To establish the method of culturing calf trabecular cells (CTCs) in vitro, to understand the morphology and function of CTCs, to probe into the effect of resistance of aqueous outflow in the pathogenesis and mechanism about primary open angle glaucoma(POAG). To direct the clinical using of drugs and to open up new antiglaucomatous medicine by pharmacological studies of CTCs.


METHODS
Trabecular meshwork was collected from twenty eyeballs of calf donors after slaughter. The tiusse was primarily cultured and cells were subcultured. The growing characteritics and morphological features of cultured primary and passaged cells were observed by light and electron microscopes. Cell kinetics of the third and tenth passaged cells were analysed using autoradiography and flow cytometry. The influence of the antiglaucomatous drugs 0.25 mg.ml-1 epinephrine (EPI) and 0.025 mg.ml-1 dipivalyl epinephrine (DPE) on cell kinetics of the third passaged cells was studied.


RESULTS
The growing characteritics and morphological features of cultured CTCs were as same as those of human trabecular cells. Growing types of CTCs included most of epitheial cell and few of fibroblast. The amount of cellular microfilaments was reduced, DNA synthesis time(Ts) and cell cycle time(Tc) were obviously prolonged with passaged increasing. Antiglaucomatous drugs-EPI (0.25 mg.ml-1) and DPE (0.025 mg.ml-1) made microfilaments dissolving, Ts and Tc obviously prolonging.


CONCLUSIONS
Establishing the method of culturing CTCs in vitro and understanding their morphology, function and pharmacological effects provided an important information for studying human trabecular cells and probing into the effect of resistance of aqueous outflow in the pathogenesis and mechanism about POAG. These studies indicated that antiglaucomatous drugs-EPI (0.25 mg.ml-1) and DPE (0.025 mg.ml-1) influenced obviously microfilaments and cell kinetics of the third passaged cells and suggested that it is not to be ignored that 1% EPI and 0.1% DPE may make CTCs' microfilaments dissolving and may inhibit CTCs' division and proliferation when they are used in clinical therapy.
Zhong
L
L
Department of Ophthalmology, First School of Clinical Medicine, Beijing Medical University, Beijing 100034, China.
Li
M
M
D008841:Actin Cytoskeleton / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure; D000818:Animals; D001082:Aqueous Humor / Q000378:metabolism; D002417:Cattle; D002478:Cells, Cultured; D003599:Cytoskeleton / Q000648:ultrastructure*; D004837:Epinephrine / Q000031:analogs & derivatives* / Q000494:pharmacology; D005902:Glaucoma, Open-Angle / Q000209:etiology*; D007700:Kinetics; D014129:Trabecular Meshwork / Q000166:cytology* / Q000648:ultrastructure
C015173:dipivefrin; D004837:Epinephrine