In vitro culture and study of the biological characteristics of rabbit keratocytes.
OBJECTIVE
To improve the technique for culturing rabbit keratocytes in vitro and investigate the biological characteristics of these cells.


METHODS
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.


RESULTS
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.


CONCLUSIONS
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.
Huo
Xiao-Kun
XK
Department of Anatomy, First Military Medical University, Guangzhou 510515, China.
Yuan
Lin
L
Huang
Tao
T
Xia
Hu
H
Yu
Lei
L
Dai
Jing-Xing
JX
Qin
Jian-Qiang
JQ
D000818:Animals; D018929:Cell Culture Techniques; D003315:Cornea / Q000166:cytology*; D011817:Rabbits; D017154:Stromal Cells / Q000502:physiology*; D023822:Tissue Engineering