[Induction of epidermal stem cells of rhesus monkey into human conjunctival epithelial cells: an in vitro experiment].
OBJECTIVE
To observe the possibility of inducing the epidermal stem cells of rhesus monkey into human conjunctival epithelial cells and to investigate the plasticity of epidermal stem cells.


METHODS
Epidermal stem cells of rhesus monkey were cultured in vitro and separated with type IV collagen. The cells were analyzed before and after isolation with beta1 integrin and keratin 15 by flow cytometry. The separated stem cells were characterized with markers, beta1 integrin and keratin 15 by immunohistochemistry and RT-PCR. The stem cells were transfected by green fluorescent protein (GFP) gene. 24 hours later, the fluorescent cells were selected out and co-cultured with human conjunctival epithelial cells in Transwell for 10 days. Then the cells were characterized with the markers of beta1 integrin, keratin 15, mucin 4 and keratin 4 by immunohistochemistry and RT-PCR.


RESULTS
The purity of the separated epidermal stem cells was almost 90%, and all the cells were positive of beta1 integrin and keratin 15. After being transfected with GFP gene for 24 hours, the cells expressed green fluorescence. 10 days later, after co-culture with human conjunctival epithelial cells, the epidermal stem cells were positive of mucin 4 and keratin 4.


CONCLUSIONS
Epidermal stem cells of rhesus monkey can be induced into conjunctival epithelial cells in vitro by co-culturing with primary passage conjunctival epithelial cells.
Lu
Rong
R
Key Laboratory of Ophthalmology, Ministry of Education, Zhongshan Ophthalmic Center of Sun Yat-sen University, Guangzhou 510060, China.
Ge
Jian
J
Huang
Bing
B
Huang
Dan-ping
DP
Gao
Nan
N
Wei
Yan-tao
YT
Wang
Zhi-chong
ZC
Peng
Zhan
Z
D000818:Animals; D002454:Cell Differentiation / Q000502:physiology; D002478:Cells, Cultured; D018920:Coculture Techniques; D003228:Conjunctiva / Q000166:cytology*; D000078404:Epidermal Cells; D004847:Epithelial Cells / Q000166:cytology*; D049452:Green Fluorescent Proteins / Q000235:genetics; D006801:Humans; D008253:Macaca mulatta; D013234:Stem Cells / Q000166:cytology*; D014162:Transfection
D049452:Green Fluorescent Proteins