基于基质胶体外构建结肠癌类器官的初步研究

王宁; 曹博; 邓欢; 崔昊; 张庆鹏; 陈润开

doi:10.3969/j.issn.2095-5227.2021.12.014

基于基质胶体外构建结肠癌类器官的初步研究

Construction of colon cancer organoids based on matrigel in vitro

摘要

摘要:
背景结肠癌类器官较传统肿瘤模型更加接近肿瘤组织的三维结构和分子特征，体外模型的构建可应用于新型抗肿瘤药物研发和肿瘤特性探究。
目的利用基质胶在体外构建出结肠癌类器官。
方法稳定培养商品化结肠癌细胞，经过消化、重悬后将其接种于基质胶内，在基质胶3D结构支撑下，培养形成具有空腔的立体球状(多叶)结构，即结肠癌类器官。光镜下连续观察结肠癌类器官的发育过程和形态特征变化；设置5×10³/mL、5×10⁴/mL、5×10⁵/mL三个密度梯度，观察类器官发育最佳密度；HE染色观察结肠癌类器官形态特点；观察抗肿瘤药物顺铂对结肠癌类器官的影响。
结果结肠癌细胞在基质胶中培养6 d后，形成多细胞球形团状结构，部分类器官发育出分叶状结构；类器官在5×10⁴/mL、5×10⁵/mL培养密度的发育情况优于5×10³/mL；顺铂在15 μg/mL浓度条件下对结肠癌类器官依然有明显的毒性效果。
结论实现了结肠癌类器官体外培养的条件探索和形态特征观察，为结肠癌的化疗药物筛选、肿瘤行为学观察等深入研究提供了良好的平台。

Abstract:
Background Compared with traditional tumor models, the colon cancer organoids are closer to the structure and molecular characteristics of tumor tissues. The model can be applied to the research and development of new anti-colon cancer drugs and the exploration of tumor characteristics.
Objective To construct organoids of carcinoma of colon with Matrigel.
Methods HCT116 cells were stably cultured and resuspended. The cells were inoculated into the Matrigel, and then cultured in three-dimensional environment to form spherical (multi-leaf) colon cancer organoids with cavities. The development and morphological characteristics of colon cancer organoids were observed and analyzed by microscope. Different inoculation gradients (5×10³/mL, 5×10⁴/mL, 5×10⁵/mL) were set to observe the appropriate density of organoids development, HE staining was used to observe the colon cancer organoids morphology, and the effect of cisplatin (DDP) on colon cancer organoids was also observed.
Results Colon cancer cells were cultured in Matrigel for 6 days to form multi-cell globular structure, and some organoids developed lobular structure. The size of the organoids grew larger at density of 5×10⁴/mL or 5×10⁵/mL were better than those at density of 5×10³/mL. In addition, the research proved that cisplatin had obvious toxic effect on colon cancer organoids.
Conclusion In this experiment, culture conditions for human colon cancer organoids are successfully established, and the morphological characteristics of colon cancer organoids in vitro are observed, which provides a more simulated model for intensive research of colon cancer.

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