Huo Xiao-Kun, Yuan Lin, Huang Tao, Xia Hu, Yu Lei, Dai Jing-Xing, Qin Jian-Qiang
Department of Anatomy, First Military Medical University, Guangzhou 510515, China.
Di Yi Jun Yi Da Xue Xue Bao. 2002 Jul;22(7):624-5.
To improve the technique for culturing rabbit keratocytes in vitro and investigate the biological characteristics of these cells.
Fresh rabbit corneas were obtained and the epithelial and endothelial cells were removed after digestion with trypsin. The stroma was rinsed, minced, and incubated in DMEM/F12 medium supplemented with 10% fetal bovine serum and the biological characteristics of the keratocytes were observed with MTT assay and compared with those of the cells in serum-free culture media.
On about the third day of incubation, some keratocytes germinated from the stromal tissues and migrated onto the flask surface presenting fibroblast-like arrangement with spindle-shaped appearance. The keratocytes became confluent after 10 day's incubation and the peak of cell proliferation occurred on day 8 in the presence of serum in the media, while in the absence of serum, the peak took place on day 10.
The method improved for in vitro keratocyte culture is convenient and effective, and the presence of serum in the media may to some degree affect the growth of the keratocytes.
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