鉴于人肝细胞原代培养在药物代谢、毒性、副作用的研究中被视为核心检验标准，而原代培养的肝细胞经常面临诸如解毒基因表达量下调的问题（如膜转运蛋白、结合酶、细胞色素P45），在Vinci B等人的肝细胞研究中（Vinci B, et al. Modular bioreactor for primary human hepatocyte culture: medium flow stimulates expression and activity of detoxification genes. Biotechnol J. 2011; 6: 554-64），认为正是传统静置培养无法模拟的某些生理刺激，引发了基因表达的下调。而Vinci B通过Quasi Vivo流动培养人原代肝脏细胞后，不论基因表达量、酶活还是生物参数，均证明流动培养可特异性地上调多种解毒酶基因的表达，而且流动培养的人贴壁肝细胞，其解毒酶基因家族的表达水平接近或高于新鲜分离的肝脏细胞。

实验流程

1、将原代肝细胞进行静态培养，在第七天分组，一组保持静态培养，一组改为Quasi Vivo流动培养，继续培养至二十一天，进行检测。

2、分别进行流动培养、静态培养、标准培养十四天后的肝脏细胞(FT297)，形态无差异  

3、流动培养下的肝细胞CYP1A1、CYP1A2、CYP2B6、CYP3A4、GSTa等解毒酶基因表达更高，到达峰值时长有差异。

4、多种解毒基因上调，CYP1A1等基因表达甚至高于新鲜分离的肝脏细胞（FIH）

5、流动培养提高了肝脏细胞CYP3A4、UGT2B4/7、CYP2C9的活性，化合物代谢速率显著提高。

文章小结：

1、人原代肝脏细胞分别进行7-21天的流动培养、静态培养；

2、检测肝脏细胞32个基因的表达量、酶活和生物参数；

3、使用流动培养可提高人贴壁肝脏细胞解毒基因表达：

①UGT（尿苷二磷酸葡萄糖醛酸转移酶）基因家族表达上调：UGT1A1/UGT2B4/UGT2B7；

②多药耐药相关蛋白1 (MDR1)和MRP2基因的mRNA的表达上调；

③外源物质/药物代谢和运输基因表达上调，并激活某些酶活性：CYP1A1/1A2/2B6/2C9/3A4；

④GST(谷胱甘肽S-转移酶)基因表达上调。

用于(贴壁)细胞培养的Quasi Vivo流动培养系统，利用培养基循环流动，模拟血流剪切应力环境，更贴近人肝脏细胞的体内环境，能促进原代肝脏细胞增殖、贴壁肝细胞的基因表达等。作为英国Kirkstall公司核心专利产品，Quasi Vivo流动培养系统创新地模拟了体内动态环境，并可同时结合3D培养构建细胞模型。流动培养系统主要由高精度蠕动泵（用于培养基循环流动）、特殊设计的培养腔室（低通量培养用、中-高通量培养用）耗材组成。Quasi Viv设备可自行拼装，操作方便，按产品说明书清洗和灭菌后，所有耗材可长期、重复使用，大大降低了Quasi Vivo的使用成本。

目前，Quasi Vivo流动培养系统已在全球超过70个专业研究机构获得应用，已成功构建包括呼吸系统（肺成纤维细胞、支气管上皮细胞等）、肝脏、肾脏、心血管、脑组织、糖尿病等研究模型（原代细胞模型）。

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