[The modification of culture and cryopreservation methods for human junctional epithelium cells].
OBJECTIVE
To study the methods for isolating, culturing and cryopreserving junctional epithelium(JE) cells.


METHODS
JE tissue samples were obtained from human periodontally healthy premolars which were extracted for orthodontic reasons. The sulcular epithelium was removed at a distance of 1 mm from the gingival margin. JE tissue was detached from the extracted teeth by using 11 sterile blade and minced into small fragments. The cultured JE cells were incubated with a freezing medium consisting of 90% calf serum and 10% DMSO. The freezing protocol was applied using a computer-controlled freezer to freeze the medium to -80 degrees centigrade before cells were plunged into liquid nitrogen and stored. The above procedures were optimized to further study the morphology, proliferation, determination of JE cells and measure the viability after thawing.


RESULTS
0.1% trypsin containing 0.02% ethylenediamine tetraacetate (EDTA) digestion was used to isolate JE cells successfully without dispase. The viability of thawing JE cells was(93.87+/-3.11)%, and the morphology was similar to that of the second passage JE cells. CK19 and vimentin were both positive in JE cells immunocytochemically.


CONCLUSIONS
The methods could be applied into practice. The phenotype of JE cells in vitro is not always consistent with that in vivo. It might be associated with the substrate which cells grow in contact with.
Shanghai Key Laboratory of Stomatology, Ninth People's Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai Research Institute of Stomatology, Shanghai 200011, China. sris@sh163.net
Shu-bo
SB
Li
De-yi
DY
Li
Xi-rui
XR
Ge
D001641:Bicuspid / Q000166:cytology*; D002478:Cells, Cultured; D015925:Cryopreservation; D004846:Epithelial Attachment; D005615:Freezing; D006801:Humans