Kagan L, Rieck P W
Klinik für Augenheilkunde, Charité - Universitätsmedizin Berlin, Campus Virchow Klinikum, Augustenburger Platz 1, 13353 Berlin.
Ophthalmologe. 2010 Apr;107(4):341-6. doi: 10.1007/s00347-009-2003-4.
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.
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.
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.
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.
原代角膜细胞（HCKp）的培养困难且受多种因素影响。本研究旨在探讨永生化角膜细胞（HCKi）在实验中是否可替代HCKp，并对角膜构建体的开发是否有用。
培养HCKp和HCKi，并分别用浓度为40 - 0.1微克/毫升的苯扎氯铵（BAC）或100 - 0.01微克/毫升的西曲溴铵孵育72小时。测量细胞活力和倍增时间（tv）。
用40或4微克/毫升BAC以及100或10微克/毫升西曲溴铵处理导致细胞死亡。HCKi的tv缩短，尤其是在用BAC处理的细胞中，但只有HCKp显示出明显的活力丧失。在用西曲溴铵处理的细胞中，两种细胞系的tv均显著增加，且从0.1微克/毫升起两种细胞系均未检测到活力丧失。
HCKi比HCKp更具抗性和增殖能力，但如果考虑到两种细胞系之间可检测到的差异，如增殖能力和对药物的反应等，在例如毒性研究等初步实验中，HCKi可作为原代细胞的替代品使用。
