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Isolation and characterization of primary canine lens epithelial cells
Author(s) -
Long Amy C.,
Agler Amanda,
Colitz Carmen M. H.,
Zhang Jin,
Hayek Michael G.,
Failla Mark L.,
Bomser Joshua A.
Publication year - 2008
Publication title -
veterinary ophthalmology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.594
H-Index - 50
eISSN - 1463-5224
pISSN - 1463-5216
DOI - 10.1111/j.1463-5224.2007.00599.x
Subject(s) - laminin , biology , cytokeratin , laboratory flask , cell culture , lens (geology) , microbiology and biotechnology , in vitro , epithelium , cell , chemistry , immunology , extracellular matrix , immunohistochemistry , biochemistry , paleontology , genetics
Objective The purpose of this study was to characterize the proliferation and differentiation of primary canine lens epithelial cells (LEC) under standard culture conditions. Procedure Canine LEC were isolated by mechanical dissection of the canine globe and enzymatic digestion of the lens capsule from fresh lenses. Isolated capsules and cell suspensions were seeded in laminin‐coated culture flasks. Canine LEC proliferated and formed monolayers, which could be passaged and maintained for approximately 2 weeks. Cells were characterized morphologically and cell lysates examined for expression of protein markers of epithelial origin and differentiation. Results Canine LEC exhibit morphologic characteristics of epithelial cells when cultured on laminin/lysine coated flasks. Expression of epithelial cell marker, cytokeratin 5, was highest at passage 1 and diminished with increasing passage number. Expression of γ‐crystallin, a protein found only in differentiated lens fiber cells, increased at passage 6. A laminin/lysine‐coated surface supported optimal proliferation of canine LEC. Both an initial seeding density of 1 × 10 5 cells/cm 2 and culture in Dulbecco's modified essential media (DMEM) supplemented with 10% FBS supported a doubling time of less than 48 h in canine LEC. Conclusion This study demonstrates that primary canine LEC retain the characteristics of lens epithelial cells prior to passage 6 under the described culture conditions and represent a suitable in vitro model for investigating lens physiology and cataractogenesis.