[The culture and cryopreservation of human conjunctival epithelium in vitro].
OBJECTIVE
To detect the best method of culture and preservation of human conjunctival epithelia.


METHODS
Human conjunctival epithelia were cultured by tissue inoculation, mechanical separation or enzyme digestion with 0.25% trypsin. The cultured cells were identified through their morphology, growing features and immunohistochemical staining. The third and fourth passage confluent cells were frozen in liquid nitrogen, and resuscitated 30 days later.


RESULTS
Cells from tissue digested with 0.25% Trypsin grew much better than those from tissue inoculation. No cell outgrew in mechanical separating group. The cultured cells spread along the flask in polygonal shape and positive in pan-Keratin staining. Ninty percent of the cryopreserved cells were successfully resuscitated.


CONCLUSIONS
Tissue digestion with trypsin is the best approach to culture human conjunctival epithelium, and the cultured cells can be cryopreserved in liquid nitrogen.
Zheng
J
J
Zhongshan Ophthalmic Center, Sun Yat-sen University of Medical Sciences, Guangzhou 510060, China.
Lu
R
R
Zhang
J
J
Lin
M
M
Lin
J
J
D018929:Cell Culture Techniques / Q000379:methods; D002478:Cells, Cultured; D003228:Conjunctiva / Q000166:cytology*; D015925:Cryopreservation / Q000379:methods; D004847:Epithelial Cells / Q000166:cytology*; D006801:Humans; D007150:Immunohistochemistry