zeta life转染原代大鼠肝细胞（IF6.684）文章分享

作者：王韵壹、李英、熊慧卿

zeta life转染原代大鼠肝细胞（IF6.684）文章分享

一、原代肝细胞转染siRNA简述

小干扰RNA转染，将原代肝细胞接种到6孔板或96孔板的含10% FBS Eagle培养基中,培养二十四几小时后，用100 nM的CHOP siRNA 或100 nM 阴性对照 siRNA (siNC) 使用美国Zeta Life公司Advanced DNA RNA转染试剂转染原代肝细胞（货号：AD600050，Zeta life，United

States）。以下CHOP siRNA序列用于转5'-CAGUAUCUUGAGUCUAAUATT-3'和 5'-UAUUAGACUCAAGAUACUGTT-3' , 转染 24 h 后，将肝细胞放置于300 μM MCT 36 h，然后对细胞活力、Annexin-V/PI 染色或蛋白质印迹测定。

培养条件：选择美国Zeta Life公司胎牛血清（＃z7181FBS-500）对原代肝细胞培养，并完成后期培养实验。

zeta life转染原代大鼠肝细胞（IF6.684）文章分享

二、Small Interfering RNA Transfection 原文分享

zeta life转染原代大鼠肝细胞（IF6.684）文章分享

Hepatocytes were seeded into 6-well plates or 96-well plates in Dulbecco’s Modifified Eagle Medium with 10% FBS. Twenty-four hours later, they were transfected with 100 nM of CHOP siRNA or 100 nM of negative control siRNA (siNC) using Advanced Transfection Reagent (Catalog No.: AD600050, Zeta life, United States) according to the manufacture’s protocol. The followingCHOP siRNA sequences were used for transfection: 5′-CAG

UAUCUUGAGUCUAAUATT-3′ (sense) and 5′-UAUUAG ACUCAAGAUACUGTT-3′ (antisense) After 24 h transfection, hepatocytes were exposed to 300 μM of MCT for another 36 h, followed by the cell viability assay, Annexin-V/PI staining, or western blotting

zeta life转染原代大鼠肝细胞（IF6.684）文章分享

三、美国Zeta Life 公司与美国加利福尼亚大学旧金山校区联合开发用于哺乳动物细胞、活体动物转染的 Advanced DNA RNA 第三代多肽小分子转染试剂，此技术成为新的全球蛋白功能、免疫细胞及干细胞治疗、研发及生产的主要关键技术之一。

四、文献中所用转染试剂产品信息

产品名称：Advanced DNA RNA 转染试剂

货号：AD600025、AD600050、AD600075、AD600150