Institute of Biochemistry, Food Science and Nutrition, The Robert H. Smith Faculty of Agriculture, Food and Environment, The Hebrew University of Jerusalem, POB 12, Rehovot 7610001, Israel.
STAR Protoc. 2020 Aug 13;1(2):100086. doi: 10.1016/j.xpro.2020.100086. eCollection 2020 Sep 18.
Primary hepatocytes are a vital tool in various biomedical research disciplines, serving as an model for liver physiology. Obtaining high yields of viable primary mouse hepatocytes is technically challenging, limiting their use. Here, we present an improved protocol based on the classic two-step collagenase perfusion technique. The liver is washed by perfusion, hepatocytes are dissociated by collagenase, separated from other cells, and cultured. This protocol was optimized to significantly reduce procedure duration and improve hepatocyte yield and viability.
原代肝细胞是各种生物医学研究领域的重要工具，可作为肝脏生理学的模型。获得高产率的活鼠原代肝细胞在技术上具有挑战性，限制了它们的使用。在这里，我们提出了一种基于经典两步胶原酶灌注技术的改良方案。肝脏通过灌注进行洗涤，胶原酶将肝细胞分离，与其他细胞分离，并进行培养。该方案经过优化，可显著缩短操作时间，提高肝细胞产量和活力。
