Establishment of lens epithelial cell lines from Emory and cataract resistant mice and their response to hydrogen peroxide.
We determined the conditions required for the establishment of lens epithelial cell lines from individual Emory and age matched cataract-resistant (CR) mice, and investigated the response of these cells to hydrogen peroxide. The technique described here permits the establishment of mouse lens epithelial cell lines from individual animals and provides an opportunity to study changes in epithelial function that precede and accompany cataract formation and aging. Capsules with lens epithelial cells were isolated from 1, 6 month, and 1.5-2 year old Emory and CR mice and cultured in minimal essential medium (MEM) containing 4% heat inactivated fetal bovine serum and 4% heat inactivated rabbit serum. Seeding efficiency at 3 hours was approximately 83% for all lines, doubling time was 22-24 hours, and the shape of the growth curves was comparable for Emory and CR mice from each age group. A three hour exposure of Emory and CR mouse lens epithelial cells from older animals to a constant level of 0.02 mM hydrogen peroxide or to an initial concentration of 0.01, 0.03, or 0.05 mM hydrogen peroxide resulted in a delay in growth. The delay in cell proliferation was decidedly more pronounced in lens epithelial cells from Emory mice. Lens epithelial cells from cataractous mice appear to be more sensitive to oxidative insult than their CR counterparts.
Reddan
J R
JR
Department of Biological Sciences, Oakland University, Rochester, Michigan 48309-4401.
Kuck
J F
JF
Dziedzic
D C
DC
Kuck
K D
KD
Reddan
P R
PR
Wasielewski
P
P
D000375:Aging; D000818:Animals; D002386:Cataract / Q000209:etiology*; D002452:Cell Count / Q000187:drug effects; D002455:Cell Division / Q000187:drug effects; D002460:Cell Line; D004848:Epithelium / Q000187:drug effects; D006861:Hydrogen Peroxide / Q000494:pharmacology*; D007908:Lens, Crystalline / Q000166:cytology / Q000187:drug effects*; D051379:Mice; D008817:Mice, Mutant Strains; D010084:Oxidation-Reduction
D006861:Hydrogen Peroxide