Purpose: The goal of this study was to examine the effects of age and long-term caloric restriction on the proliferation capacity of murine lens epithelial (LE) cells in vitro and in vivo.
Methods: B6D2F1 (C57BL/6 X DBA/2) F1 mice 4 to 45 months of age were obtained and fed either an ad libitum (AL) or a calorically restricted (CR) diet (60% of AL intake). Cellular proliferation capacity in vitro was measured using the colony size distribution assay for 10-day clonal growth of mouse LE cells. Proliferation rate in vivo was assayed using immunostaining for 5-bromo-2'-deoxyuridine (BrdU) in mouse LE cells after 2-week osmotic pump delivery of BrdU.
Results: Proliferative capacity of cells from old AL mice decreased significantly in comparison to cells from young AL and old CR mice, as determined by the fractions of cells capable of forming small (no or one cell division) and large (four or more cell divisions) colonies in vitro. There was also a decline in cell replicative rate as measured by BrdU labeling index (LI) in vivo with increasing age in AL and CR mice. However, this decline was marked in AL mice between 10 and 30 months of age and minimal in CR mice. Significant differences in BudU LI between AL and CR mice occurred when animals were 30 months of age or older. This finding indicates that an age-related decline in cellular proliferation rate in vivo was delayed by CR.
Conclusions: A significantly reduced proliferative capacity of LE cells is associated with increased age of mice and is delayed by long-term caloric restriction as measured in vitro and in vivo. How caloric restriction mediates its effects on LE cell proliferation remains to be investigated further.