Isolation and characterization of primary canine lens epithelial cells

Veterinary Ophthalmology
Amy C LongJoshua A Bomser

Abstract

The purpose of this study was to characterize the proliferation and differentiation of primary canine lens epithelial cells (LEC) under standard culture conditions. 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. 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 gamma-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 x 10(5) cells/cm(2) and culture in Dulbecco's modified essential media (DMEM) supplemented with 10% FBS supported a doubling time of less th...Continue Reading

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Citations

Nov 16, 2012·Journal of Biosciences·Anshul I AroraAbhay R Vasavada
Apr 18, 2009·Nutrition and Cancer·Elizabeth A Clubbs, Joshua A Bomser
Sep 29, 2012·Research in Veterinary Science·M D PratheeshG T Sharma
Feb 23, 2011·Chemico-biological Interactions·Chad BrockerVasilis Vasiliou
Nov 7, 2008·Journal of Veterinary Diagnostic Investigation : Official Publication of the American Association of Veterinary Laboratory Diagnosticians, Inc·Francesco SassiCinzia Benazzi
Feb 2, 2012·Experimental Animals·Nobuyuki KanemakiHideharu Ochiai

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