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Isolation of primary open-angle glaucomatous trabecular meshwork cells from whole eye tissue.

Publication ,  Journal Article
Stamer, DW; Roberts, BC; Epstein, DL; Allingham, RR
Published in: Curr Eye Res
May 2000

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 (3 of 4 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. CONCLUSION: 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.

Duke Scholars

Published In

Curr Eye Res

ISSN

0271-3683

Publication Date

May 2000

Volume

20

Issue

5

Start / End Page

347 / 350

Location

England

Related Subject Headings

  • Trabecular Meshwork
  • Ophthalmology & Optometry
  • Humans
  • Glaucoma, Open-Angle
  • Collagenases
  • Cell Separation
  • Cell Culture Techniques
  • Aged, 80 and over
  • Aged
 

Citation

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Stamer, D. W., Roberts, B. C., Epstein, D. L., & Allingham, R. R. (2000). Isolation of primary open-angle glaucomatous trabecular meshwork cells from whole eye tissue. Curr Eye Res, 20(5), 347–350.
Stamer, D. W., B. C. Roberts, D. L. Epstein, and R. R. Allingham. “Isolation of primary open-angle glaucomatous trabecular meshwork cells from whole eye tissue.Curr Eye Res 20, no. 5 (May 2000): 347–50.
Stamer DW, Roberts BC, Epstein DL, Allingham RR. Isolation of primary open-angle glaucomatous trabecular meshwork cells from whole eye tissue. Curr Eye Res. 2000 May;20(5):347–50.
Stamer, D. W., et al. “Isolation of primary open-angle glaucomatous trabecular meshwork cells from whole eye tissue.Curr Eye Res, vol. 20, no. 5, May 2000, pp. 347–50.
Stamer DW, Roberts BC, Epstein DL, Allingham RR. Isolation of primary open-angle glaucomatous trabecular meshwork cells from whole eye tissue. Curr Eye Res. 2000 May;20(5):347–350.
Journal cover image

Published In

Curr Eye Res

ISSN

0271-3683

Publication Date

May 2000

Volume

20

Issue

5

Start / End Page

347 / 350

Location

England

Related Subject Headings

  • Trabecular Meshwork
  • Ophthalmology & Optometry
  • Humans
  • Glaucoma, Open-Angle
  • Collagenases
  • Cell Separation
  • Cell Culture Techniques
  • Aged, 80 and over
  • Aged