猪结肠脱细胞支架联合结肠癌HCT116细胞的体外共培养  

In vitro co-culture of decellularized matrix of porcine colon with colon cancer HCT116 cells

作　　者：朱茗祺 周京旭[2] 林丽珠[3] 陈泽仁 廖志晓 Zhu Mingqi;Zhou Jingxu;Lin Lizhu;Chen Zeren;Liao Zhixiao(Guangzhou University of Chinese Medicine,Guangzhou 510006,Guangdong Province,China;District 1,the First Affiliated Hospital of Guangzhou University of Chinese Medicine,Guangzhou 510400,Guangdong Province,China;District 4,Cancer Center,the First Affiliated Hospital of Guangzhou University of Chinese Medicine,Guangzhou 510400,Guangdong Province,China)

机构地区：[1]广州中医药大学,广东省广州市510006 [2]广州中医药大学第一附属医院肿瘤中心一区,广东省广州市510400 [3]广州中医药大学第一附属医院肿瘤中心四区,广东省广州市510400

出　　处：《中国组织工程研究》2021年第10期1533-1538,共6页Chinese Journal of Tissue Engineering Research

摘　　要：背景:近年来脱细胞支架已被广泛用于各种肿瘤的研究,支架的空间排列、生物力学性质和生物相容性等特性均有助于还原肿瘤细胞生长的微环境。目的:探讨猪结肠脱细胞支架作为结肠癌体外模型的特点与优势。方法:将新鲜猪结肠以2%SDS、1%Triton X-100及0.5%EDTA浸泡结合反复振荡的方法进行脱细胞处理,制备猪结肠脱细胞支架。将人结肠癌HCT116细胞接种于猪结肠脱细胞支架黏膜面,活-死染色观察支架上的细胞生长情况,鬼笔环肽染色观察支架上的细胞形态,苏木精-伊红染色与免疫荧光染色观察细胞在支架上的纵向生长情况。结果与结论:①活-死染色显示,培养第1天,HCT116细胞能很好地聚集在支架黏膜层的孔隙内,少见死细胞;第3天时,细胞逐渐向孔隙外蔓延生长,小部分细胞已生长连接成片,无死细胞;第7天时,细胞生长密度进一步增大,在黏膜层表面生长成片,无死细胞与脱落细胞;②培养第7天苏木精-伊红染色及免疫荧光染色显示,部分HCT116细胞可在孔隙内生长成团,并有部分细胞沿着孔隙继续向黏膜下层生长,表现出原位肠癌浸润性生长的特点;③培养第7天鬼笔环肽染色显示,HCT116细胞与支架黏膜层在超微结构水平上有紧密的接触,并且具有分化良好的上皮形态特征;④结果表明,猪结肠脱细胞支架可作为结肠癌细胞三维培养的载体。BACKGROUND: Acellular scaffolds have been widely used in the study of various tumors in recent years. The spatial arrangement, biomechanical properties and biocompatibility of the scaffolds are helpful to restore the microenvironment of tumor cell growth.OBJECTIVE: To investigate the characteristics and advantages of porcine colon acellular scaffold as an in vitro model of colon cancer.METHODS: Acellular scaffolds of porcine colon were prepared by soaking fresh porcine colon with 2% SDS, 1% TritonX-100 and 0.5% EDTA combined with repeated shaking. Human colon cancer HCT116 cells were inoculated on the mucous surface of porcine colon acellular scaffold. The growth of cells on the scaffold was observed by live-death staining. The morphology of cells on the scaffold was observed by phalloidin staining, and the longitudinal growth of cells on the scaffold was observed by hematoxylin-eosin staining and immunofluorescence staining.RESULTS AND CONCLUSION:(1) Live-death staining showed that on the first day of culture, HCT116 cells could well gather in the pores of the mucosal layer of the scaffolds, and few dead cells were found. On the third day, the cells gradually spread out of the pores, and a small number of cells grew and connected into pieces, and there were no dead cells. On the 7 th day, the cell growth density further increased and grew into a sheet on the surface of the mucous layer, and there were no dead cells and exfoliated cells.(2) On the 7 th day of culture, hematoxylin-eosin staining and immunofluorescence staining showed that some HCT116 cells could grow into clusters in the pores, and some cells continued to grow to the submucosa along the pores, showing the characteristics of invasive growth of intestinal cancer in situ.(3) On the 7 th day of culture, the phalloidin staining showed that HCT116 cells were in close contact with the scaffold mucosa at the ultrastructural level, and had well-differentiated epithelial morphological characteristics.(4) The result shows that porcine colon acellular scaffol

关 键 词：材料 肿瘤 支架 脱细胞支架 三维培养 结肠癌 浸润性生长 

分 类 号：R459.9[医药卫生—治疗学] R318.08[医药卫生—临床医学]