Establishment of a corneal epithelial cell line spontaneously derived from human limbal cells.
The objective of this study was to establish a spontaneously derived human corneal epithelial cell line from a normal human limbus that retains differentiation potential and proliferative properties under continuous cell culture. After 50 passages of epithelial cells obtained from human limbal tissue a cell line spontaneously emerged. The immortalized cells showed a cobblestone appearance and displayed dense microvilli on their apical cell surface membrane. Colony forming efficiency was 5-6% and population doubling time was 19.6 h. In the mRNA level, cytokeratin (CK) 3 and 12 were detected in this cell line. In the protein level, the cells expressed CK3, CK12, CK14, CK19, vimentin, and some other proteins such as F-actin and beta-tubulin and beta(1)-integrin. They lacked p63. The immortalized cells had a heteroploid karyotype, but did not exhibit tumorigenic features. When cultured on an air-liquid interface the cells could form stratified multilayer epithelia. In summary, all these results indicated that a new human corneal epithelial cell line was spontaneously established from normal limbal tissue through serial culture. This cell line would be useful for studies of corneal epithelial biology and reconstructive corneal tissue engineering.
Liu
Jingbo
J
State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University, 54 Xian Lie Nan Road, Guangzhou, Guangdong 510060, China.
Song
Ge
G
Wang
Zhichong
Z
Huang
Bing
B
Gao
Qianying
Q
Liu
Bingqian
B
Xu
Ying
Y
Liang
Xuanwei
X
Ma
Ping
P
Gao
Nan
N
Ge
Jian
J
D015153:Blotting, Western / Q000379:methods; D018929:Cell Culture Techniques; D002453:Cell Cycle; D018797:Cell Cycle Proteins / Q000032:analysis; D002460:Cell Line; D049109:Cell Proliferation; D002471:Cell Transformation, Neoplastic; D015925:Cryopreservation; D004847:Epithelial Cells; D019573:Epithelium, Corneal / Q000166:cytology*; D006801:Humans; D007621:Karyotyping; D008032:Limbic System / Q000166:cytology*; D008854:Microscopy, Electron; D008858:Microscopy, Phase-Contrast; D013234:Stem Cells
D018797:Cell Cycle Proteins