制备和培养多细胞肿瘤球体的简易方法

Simple Method for Preparation and Culture of Multi-cellular Tumor Spheroid

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 。

Objective To find a rapid and simple method for preparation of multi-cellular tumor spheroid (MTS). Methods Two cell lines of pulmonary adenoma (AGZY and Anip) in our department were prepared for MTS with agar culture method of tuning bottle. Results The many convergencies of small amount cells were formed at 12~24 h after culture and then their outer layer cells division and proliferation were taken to form the spheroid in identical size and regular shape. MTS increased gradually and entered the platform stage after 12 days its surface cells dropped in 15 d and MTS collapsed. Conclusion This method could induce the strongly adhibit-wall cell lines of AGZY and Anip to form MTS. The quantity of MTS might be controlled with the cell concentration and MTS were unanimous in size and regular in shape. This method could be used in general lab because it could be simple and convenient for observation without higher requirement of equipment.