Weisser H, Witzel J G, Lathan B
Institute of Clinical Chemistry and Laboratory Medicine, University Clinic Bergmannsheil, Bochum, Germany.
Tumour Biol. 1994;15(3):153-9. doi: 10.1159/000217886.
The conventional human tumor stem cell assay is limited by the lack of flexibility for drug scheduling with single agents and its inability to test drug combinations. Recently, we described the cloning of tumor cell lines within porous glass capillary tubes which allow free exchange of substances. The present study describes the influence of various perfusion modalities on the colony growth of the human tumor cell line MDA-231 cloned within the perfused capillary cloning system (PCCS). Colony growth of tumor cells within the PCCS is strongly dependent on perfusion tube volume, flow rate and duration of perfusion with growth medium. Best colony growth was achieved using a perfusion tube volume of 12 ml resulting in a cloning efficiency of 36.3%. Continuous perfusion with fresh medium did not improve the cloning efficiency; in fact, colony growth was hampered compared to colony growth within unperfused porous capillaries. However, cloning efficiency was acceptable when continuous perfusion was started at day 6 (26.4%) instead of day 0 (17.2%), or when a short perfusion with high volume (12 ml/h) was discontinued after 1 h at day 0. In contrast to the conventional capillary cloning system the PCCS has the potential for investigating secretion and kinetics of tumor-specific factors and the effect of growth-stimulating or growth-inhibiting drugs.
传统的人类肿瘤干细胞检测方法存在局限性，即单药给药方案缺乏灵活性，且无法检测药物组合。最近，我们描述了在多孔玻璃毛细管内克隆肿瘤细胞系的方法，这种毛细管允许物质自由交换。本研究描述了各种灌注方式对在灌注毛细管克隆系统（PCCS）内克隆的人类肿瘤细胞系MDA - 231集落生长的影响。PCCS内肿瘤细胞的集落生长强烈依赖于灌注管体积、流速和生长培养基的灌注持续时间。使用12 ml的灌注管体积可实现最佳集落生长，克隆效率为36.3%。用新鲜培养基持续灌注并不能提高克隆效率；事实上，与未灌注的多孔毛细管内的集落生长相比，集落生长受到了阻碍。然而，当在第6天（26.4%）而非第0天（17.2%）开始持续灌注时，或者在第0天进行1小时的高体积（12 ml/h）短时间灌注后停止灌注时，克隆效率是可以接受的。与传统的毛细管克隆系统不同，PCCS有潜力研究肿瘤特异性因子的分泌和动力学以及生长刺激或生长抑制药物的作用。
