H460小型细胞团在天然活性Ⅰ型胶原中三维立体培养模型的构建  

Three-dimensional culture of H460 small cell clumps in native type Ⅰ collagen gel

作　　者：肖永久[1] 朱运奎[1] 汤育瑛[1] 李继东[1] 薛庆亮[1] 刘卫[1] 王晓芹[1] 牟玉兰[1] 

机构地区：[1]解放军兰州军区兰州总医院呼吸内科,甘肃省兰州市730050

出　　处：《中国组织工程研究与临床康复》2007年第41期8382-8384,共3页Journal of Clinical Rehabilitative Tissue Engineering Research

基　　金：甘肃省自然科学基金项目(3ZS051-A25-088)~~

摘　　要：目的:建立H460小型细胞团在天然活性Ⅰ型胶原中的三维立体培养技术。方法:实验于2006-07/11在解放军兰州军区兰州总医院呼吸内科实验室(博士后工作站)完成。①实验材料:H460细胞系由解放军总医院惠赠;Wistar大鼠,体质量150～200g,实验中的胶原仅从大鼠的尾部肌腱提取,该大鼠来源为其他科室实验过程中未做任何处理的大鼠。②实验方法:将H460小型细胞团悬液、鼠尾提取的天然胶原(4g/L)与4倍体积的DMEM共同混合为细胞浓度2×108L-1、胶原浓度为1g/L、1倍体积DMEM浓度。③实验评估:倒置显微镜观察细胞团在天然胶原中的生长状态,并与平面培养细胞相比较。结果:平面培养的细胞,其细胞团生长呈分散平铺的方式,单个细胞为不规则多边行,细胞之间有明显的间隙存在,而立体培养的细胞呈团块状,周围可见放射状毛刺突起,无法观察单个细胞。结论:H460小型细胞团在天然胶原中的三维立体培养为体外研究细胞生长及肿瘤细胞生长方式提供了一个更为类似体内环境的模型。AIM： To establish the model of three-dimensional culture of H460 small cell clumps in native type I collagen gel. METHODS： From July to November in 2006, the experiment was accomplished in the Laboratory of Respiratory Medicine （Post-doctoral Station）, Lanzhou General Hospital of Lanzhou Mititary Area Command of Chinese PLA.（1）H460 small cell clumps ware offered by General Hospital of Chinese PLA as a gift, and ware extracted from Wistar rats weighing 150-200 g. The native type Ⅰ collagen was extracted from rat-tail tendons.（2）The native collagen （4 g/L） was mixed with four-fold concentrated DMEM and H460 cell suspension so that the final mixture resulted in 1 g/L of collagen, about 2×10^8 cells/L, one-fold concentrated DMEM. Inverted microscope was used to observe the growing morphology of cell clumps in native collagen gels and compare with the cells cultured on plastic plate. RESISTS： The proliferation of H460 cell clumps in collagen gels was dimensional and the cell lumps broke out radic burrs. On the contrary, cells spread out the monolayer cultured clumps in plate were irregular and multilateral. The cell-cell intervals could be seen under microscope. CONCLUSION： Three-dimensional culture of H460 small cell clumps using a native type Ⅰ collagen gel provides an environment more similar to in vivo conditions for investigating the vegetal way of cancer cells in vitro.

关 键 词：活性Ⅰ型胶原 三维立体培养 H460 组织构建 

分 类 号：R318[医药卫生—生物医学工程]