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Updated: Aug 7, 2026

Optimizing Mouse Primary Lens Epithelial Cell Culture: A Comprehensive Guide to Trypsinization
Published on: June 21, 2024
[Effects of calcium on lens epithelial cells in rabbits]
Koichiro Mukai1, Hiroyuki Matsushima, Yasuo Ishii
1Department of Ophthalmology, Dokkyo University School of Medicine, Tochigi, Japan. k.mukai@jcom.home.ne.jp
Purpose:
The action of lens epithelial cells (LECs) is important for cataract and posterior subcapsular cataract after cataract surgery. In this study, we analyzed the effects of calcium on the characteristics of LECs.
Methods:
The LECs were collected using albino rabbits and incubated in minimum essential medium [MEM, Introgen Corp. (12% fetal bovine serum: FBS)] (37 degrees C, 5 % CO2) for a week to induce their proliferation. Cell culture dishes (35 mm) were prepared and 7 mm cylindrical pipes were placed in them. After that, around 10,000 cultured LECs were placed in the pipes and incubated. After 2 hours incubation, the pipes were removed and various doses of MEM (0, 2, 10 and 20 mM) replaced the calcium. Proliferation and shapes of LECs were observed using a confocal microscope and immunohistological analysis [alpha-smooth muscle actin (alpha-SMA) and bromodeoxyuridine (BrdU)]. The LECs were incubated with collagen gel and different calcium doses (0, 2, 10 and 20 mM) of MEM to calculate the contraction rate.
Results:
It was observed that the LECs changed to fibroblast-like cells at high doses of calcium using a confocal microscope. Histological studies showed that the BrdU positive cells were increased by using 10 and 20 mM calcium MEM, but the positive cells were decreased by using 0 and 2 mM calcium MEM. Increase of alpha-SMA stained cells was recognized when using 0, 10 and 20 mM calcium MEM. The contraction rate of collagen gel was increased by using the 10 and 20 mM calcium MEM.
Conclusion:
The changes of calcium concentration might be an important factor for the development of cataract, posterior subcapsular opacification, and contraction of the lens capsule after cataract surgery.