[A comparative in vitro analysis of primary and immortalized keratocytes].
BACKGROUND
The cultivation of primary keratocytes (HCKp) is difficult and influenced by a multitude of factors. In this study it was examined if immortalized keratocytes (HCKi) can replace HCKp in experiments and be useful in the development of a cornea construct.


METHODS
HCKp and HCKi were cultivated and incubated for 72 h with benzalkonium chloride (BAC) or cetrimide at concentrations of 40-0.1 microg/ml or 100-0.01 microg/ml. The vitality and the doubling time (tv) were measured.


RESULTS
Treatment with 40 or 4 microg/ml BAC as well as 100 or 10 microg/ml cetrimide led to cell death. The tv was shortened in HCKi especially in cells that were treated with BAC, but only HCKp showed a significant loss of vitality. In cells treated with cetrimide the tv increased significantly in both cell lines and no loss of vitality was detected from 0.1 microg/ml onwards in both cell lines.


CONCLUSIONS
HCKi are more resistant and proliferative than HCKp but they can be used in preliminary experiments as an alternative to primary cells in for example toxicity studies if the detectable differences between the two cell lines, such as the capacity for proliferation and reaction to agents are taken into consideration.
Kagan
L
L
Klinik für Augenheilkunde, Charité - Universitätsmedizin Berlin, Campus Virchow Klinikum, Augustenburger Platz 1, 13353 Berlin. L.Kagan@web.de
Rieck
P W
PW
D049109:Cell Proliferation; D002470:Cell Survival; D002478:Cells, Cultured; D003319:Corneal Stroma / Q000166:cytology*; D005347:Fibroblasts / Q000145:classification* / Q000166:cytology*; D006801:Humans