3株肝细胞在硝酸纤维素膜界面上生长的初步研究  

Preliminary study of three liver cells cultured on organic interface of nitrocellulose membrane

作　　者：王亚东[1] 李爱民[1] 朱薇[1] 燕群[1] 邓丽娟[1] 鲁春花[2] 刘址忠[1] 刘思德[1] 

机构地区：[1]南方医科大学南方医院省部共建器官移植重点实验室,510515 [2]南方医科大学基础医学院医学检验系,510515

出　　处：《现代消化及介入诊疗》2011年第2期96-100,104,共6页Modern Interventional Diagnosis and Treatment in Gastroenterology

基　　金：国家863计划(2006AA02A140);南方医院院长基金(2009C016)

摘　　要：目的在体外培养的条件下,观察3株肝细胞在硝酸纤维素膜材料上的生长情况,研究膜的细胞相容性及理化性能的改变,初步探讨硝酸纤维素膜作为一种新的组织工程膜材料用于构建生物人工肝生物反应器中细胞的载体材料的可行性。方法将生物人工肝常用的细胞:肝肿瘤细胞HepG2、C3A、永生化HL7702细胞株分别接种在浸泡处理后的硝酸纤维素膜片上常规培养,在普通光学显微镜以及扫描电镜观察肿瘤细胞在此种膜上的贴壁情况和细胞形态。同时行免疫细胞化学染色和细胞功能检测。结果细胞经常规染色、透明处理后,在光镜下观察到膜上生长的细胞保持了较好的增殖状态及细胞形态;免疫细胞化学染色结果表明,膜上细胞特异性标志物表达良好。培养液上清的生化检测表明培养细胞的主要分泌、代谢功能与常规培养无明显差异;扫描电镜下膜表面细胞呈集落式三维生长,增殖明显。结论硝酸纤维素膜能促进细胞黏附,并且与细胞有良好的相容性。硝酸纤维素膜作为一种良好的细胞黏附生长介质的同时,还具有可以在其上面直接进行相关培养物观察和检测的优点。Objective The purpose of this study is to test the biological compatibility of NC（nitrocellulose） membrane with liver cells in vitro.Methods Three liver cell lines （HepG2,C3A,and immortalization cell line HL7702） were cultured on the pretreatment NC membrane,cell adhesion and cell morphology were observed by optical microscopy and scanning electron microscopy,and the cell growth and biological functions were detected by immunocytochemistry.Results After transparent treatment and staining,cells on NC membrane observed under light microscope maintained better cell proliferation and cell morphology.IHC data indicated that the specific markers of cells on membrane expressed normally.The biochemistry detections of cell culture supernatant indicated that the secretion and metabolic functions of cells on NC membrane were similar with conventional culture.Scanning electron microscope showed that cell colony on membrane grew in threedimensional pattern and proliferated significantly.Conclusion NC membrane could promote cell adhesion,and had good histocompatibility.NC membrane was not only a good biological carrier in cell growth,but also an useful material for morphology observation and cell function tests in situ.

关 键 词：硝酸纤维素膜 HEPG2细胞株 C3A细胞株 HL7702细胞株 免疫细胞化学染色 组织相容性 

分 类 号：R446.61[医药卫生—诊断学]