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目的 通过实验找出一种更加简便、快捷的制备MTS的方法。材料和方法 利用本实验室的 2个肺腺癌细胞系AGZY和Anip ,通过转瓶琼脂培养法 (即在培养瓶底部涂上 1～ 2mm厚的琼脂层 ,使之凝固后接种一定浓度的细胞悬液 ,置于 37℃、5 0r/min的恒温摇床上旋转培养 )进行MTS的制备和培养。结果 经过 12～ 14h后 ,即可见许多较小的细胞集聚体形成 ,以后集聚体外层细胞不断分裂增生 ,逐渐形成大小较一致、形态较规则的球体 ,即MTS。MTS逐渐增大 ,约 12d后生长进入平台期 ,约 15d后MTS表层细胞有脱落 ,最后MTS崩解。结论 本实验能够诱导贴壁性很强的细胞 (AGZY和Anip)形成MTS ,并且可以通过控制接种的细胞浓度来制备一定数量的MTS ;形成的MTS大小较一致 ,形态较规则。重要的是本实验操作简单 ,观察方便 ,设备要求不高 ,更便于普通实验室开展工作
Objective To find out a more simple and quick way to prepare MTS through experiments. Materials and Methods Two lung adenocarcinoma cell lines, AGZY and Anip, in our laboratory were used to pass the flask agar culture method (ie, a layer of 1-2 mm thick agar was applied to the bottom of the culture flask to solidify and inoculate a certain concentration of cells. The suspension was placed on a constant temperature shaker at 37° C. and 50 r/min to prepare and culture MTS. Results After 12 to 14 hours, many small cell aggregates can be seen. Afterwards, the cells in the outer layer of the aggregating cell continuously divide and proliferate, and gradually form a sphere of uniform size and regular shape, namely MTS. The MTS gradually increased and grew to the plateau stage after about 12 days. After about 15 days, the MTS superficial cells were detached. Finally, the MTS disintegrated. Conclusion This experiment can induce the formation of MTS in cells with strong adhesion (AGZY and Anip), and can control the number of MTS by inoculating the cell concentration; the formed MTS size is relatively uniform and the morphology is more regular. The important thing is that this experiment is simple, easy to observe, and the equipment does not require much, which is more convenient for ordinary laboratories to carry out their work.