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Abstract #5984 Published in IGR 2-2
Isolation of primary open-angle glaucomatous trabecular meshwork cells from whole eye tissue
Stamer DW; Roberts BC; Epstein DL; Allingham RRCurrent Eye Research 2000; 20: 347-350
PURPOSE: Isolation and culture of human trabecular meshwork (TM) cells from primary open-angle glaucomatous (POAG) tissue has proven difficult. The objective of this study was to directly compare the utility of two different isolation methods to obtain viable human TM cells from POAG whole eye tissue. METHODS: Using a blunt dissection technique, human TM tissue was obtained from four pairs of donor eyes (67, 77, 81 and 82 years) with a documented history of POAG. TM tissue from one eye was explanted into tissue culture. TM from the contralateral eye was digested with a collagenase mixture and seeded onto culture plates. RESULTS: Primary cell isolates were obtained from all donors with both techniques. However, only cells obtained using the digestion method (three of four TMs) could be passaged for expansion and freeze-downs (3 x 107 second passage cells/donor). None of the cells obtained from explanted TMs could be passaged. Cells from successful isolations were of uniform size, possessed typical TM morphology and had doubling times <48 hours. CONCLUSIONS: These results demonstrate a clear advantage to digesting the extracellular matrix of glaucomatous TM tissue to obtain sufficient numbers of healthy cells for use in experiments. In contrast to cells obtained from explants, cells liberated from POAG TM tissue by digestion appear indistinguishable morphologically and behaviorally from 'normal' TM cells.
Dr. D.W. Stamer, Departments of Ophthalmology and Pharmacology, The University of Arizona, Tucson, AZ 85719, USA. dstamer@eyes.arizona.edu
Classification:
2.5 Meshwork (Part of: 2 Anatomical structures in glaucoma)
4 Tissue culture of ocular cells
